9th OCARINA International Symposium
Joint mini-symposiums on Structural Biology and Catalysis

Preface
The OCU Advanced Research Institute for Natural Science and Technology(OCARINA) is organized to ﬁnd solutions for global-level energy and environmental
issues and complex and advanced research subjects, and contribute to the establishmentof a sustainable society. Since its foundation in 2010, OCARINA has set up research
projects across the graduate schools to promote multidisciplinary researches.
Currently, we have seven big projects from Graduate School of Science, Graduate
School of Engineering, Graduate School of Human Life Science and Graduate School of
Medicine in Osaka City University (OCU). To promote interdisciplinary discussions
among those research projects, we will hold the 9th international symposium on
March 6-7, 2018, at OCU Osaka, Japan.
This symposium organizes the annual reports of the research projects andinvited lectures. We added two keynote subjects on the structural biology by using
cryo-electron microscopy, which honored the 2017 Nobel Prize in chemistry, andcatalysis, which is a crucial substrate to govern chemical reaction.
The symposium promotes the poster presentations for younger carrier
researchers (age under 35) in order to increase opportunities for lively and informative
exchange of views. Thanks for the promotions; we successfully received over 60
submissions. Approximately 20% of posters will be awarded the poster prize. It will
be announced at the banquet.
On behalf of organizing committee, it is my great pleasure to welcome you tothis symposium and I hope all of you to interact and enjoy with fruitfull discussions.
Chair organizer

Michio MIYANOAdviser to the President of OCU
Director of OCARINA

1

第９回 OCARINA 国際シンポジウム
ご挨拶大阪市立大学・複合先端研究機構（OCARINA）は、地球規模でのエネルギー、資源、生態系など、環境を含めた全人類に係る複合的および先端的な研究課題に対して、プロジェク
ト制により研究科横断型で最先端科学・技術を融合して取り組むことにより、学術的・社会的提言並びに人材育成を行い、得られた成果を社会や地域へ効果的に還元することを目的とします。
現在、機構では理学研究科・工学研究科・生活科学研究科の３研究科を横断する研究組織を構築し、７つのプロジェクト研究を推進しています。これらのプロジェクト研究の相互理解を深める場として、「第９回OCARINA国際シンポジウム」を開催いたします。
今年度は、2017年ノーベル化学賞を受賞した「クライオ電子顕微鏡」と、化学反応をつ
かさどる極めて重要な物質である「触媒」に関する２つのトピックスについて、研究のトップランナーとしてご活躍の外国人研究者による講演を企画しました。
また、学内理系分野間の相互理解を深めることを目的として、本学「人材育成プロジェ
クト」と連携したポスターセッションを設けました。おかげさまで、60名を超える申し込
みをいただきました。
ポスターは英語によるショートプレゼンテーションとポスター発表
により審査され、優秀ポスター賞が贈られます。 ポスター賞授与式は懇親会で行います。
ご参加された皆様にはぜひ活発な異分野交流と議論を行っていただけますようお願い申し上げます。

2018年３月
複合先端研究機構・機構長
宮野

2

道雄

Date
6th March (Tue.) , 2018 (Registration:09:00-) 09:30-18:00
7th March (Wed) , 2018 (Registration:09:30-) 10:00-18:00*Notice: Symposium Photo at 10F (18:10-18:20)

Venue
Conference Room (10th Floor) , Media Center, OCU

Banquet7th March (Wed.) , 2018 18:30at "Nonohana House" 1st Floor, Media Center, OCU

Organizer
The OCU Advanced Research Institute for Natural Science and Technology (OCARINA)

Organizing committee
Dr. Michio MIYANO (Adviser to the President of OCU / Director of OCARINA/Professor, Graduate School of Human Life Science, OCU, Japan)
Dr. Nobuo KAMIYA (Vice Director, OCARINA/Professor, OCARINA, Japan)
Dr. Naoteru SHIGEKAWA (Vice Director, OCARINA/Professor, Division of Physical Electronics and Informatics (Applied Physics andElectronics), Graduate School of Engineering, OCU, Japan)
Dr. Yutaka AMAO (Director, ReCAP/Professor, OCARINA, Japan)
Dr. Tomoko YOSHIDA (Deputy Director, ReCAP/Professor, OCARINA, Japan)
Dr. Ritsuko FUJII (Associate Professor, OCARINA, Japan)
Dr. Keisuke KAWAKAMI (Specially Appointed Associate Professor, OCARINA, Japan)
Dr. Tomoyasu NOJI (Specially Appointed Lecturer, OCARINA, Japan)
Dr. Shusaku IKEYAMA (Specially Appointed Research Associate, OCARINA, Japan)
Dr. Yoshihiro YAMAGUCHI (Tenure-track Specially Appointed Associate Professor, OCARINA, Japan)

Invited speakers
Dr. Pu QIAN (Researcher, The Univ. of Sheffield, United Kingdom)
Dr. Laurean ILIES (Associate Professor, Graduate School of Science, The Univ. of Tokyo, Japan)
Dr. Xing Yi LING (Associate Professor, Nanyang Technological Univ., Singapore)
Dr. Katsura SUGAWARA (Japan Tissue Engineering Co. Ltd.)
Dr. Mitsuhiro TERAKAWA (Associate Professor, Faculty of Science and Technology, Keio Univ., Japan)
Dr. Tasuku HAMAGUCHI (Researcher, RIKEN SPring-8, Japan)
Dr. Sin-ichiro OZAWA (Specially appointed Research Associate, Research Institute for Interdisciplinary Science, Okayama Univ., Japan)
Dr. Masahiro UEDA (Professor, Graduate School of Frontier Biosciences, Osaka Univ., Japan)
Speakers in OCARINA project teamDr. Taro NAKAMURA（Professor, Division of Biology and Geosciences(Functional Biosciences), Graduate School of Science,OCU, Japan）
Dr. Soichi SAEKI (Associate Professor,Division of Mechanical and Physical Engineering (Mechanical Engineering), Graduate School ofEngineering, OCU,Japan)
Dr. Tatsuya SHOJI (Lecturer, Graduate School of Science, Division of Molecular Materials and Science(Inorganic Chemistry),
OCU,Japan)
Dr. Yasuhiro NAKASO (Special Researcher, OCARINA, OCU,Japan)
Dr. Masaki NAKAO (Extraordinary Professor, OCARINA, OCU,Japan)
Dr. Tomoko YOSHIDA (Deputy Director, ReCAP/Professor, OCARINA, Japan)
Dr. Makoto MIYATA (Professor, Division of Biology and Geosciences(Functional Biosciences), Graduate School of Science, OCU, Japan)
Dr. Akihisa TERAKITA (Professor, Division of Biology and Geosciences(Science of Biomolecules), Graduate School of Science, OCU, Japan)
Dr. Tetsuya SATOH (Professor, Division of Molecular Materials and Science(Organic Chemistry), Graduate School of Science OCU, Japan)
Dr. Naoteru SHIGEKAWA (Deputy Director, OCARINA/Professor, Division of Physical Electronics and Informatics (Applied Physics andElectronics), Graduate School of Engineering, OCU, Japan)

3

日

時

2018年3月6日（火）（受付9:00） 9:30−18:00
2018年3月7日（水）（受付9:30） 10:00−18:00＊3月7日終了後、会場にて集合写真を撮りますのでご参集ください。
会場

大阪市立大学（杉本キャンパス）学術情報総合センター 10階 大会議室 研究者交流室
大阪市住吉区杉本3丁目3番138号 Tel:06-6344-9560
懇親会
2018年3月7日（水）18:30〜
学術情報総合センター１階
主

カフェレストラン「野のはなハウス 」

催

大阪市立大学複合先端研究機構
組織委員
宮野 道雄
（大阪市立大学・学長補佐／大阪市立大学・複合先端研究機構・機構長／生活科学研究科・教授）
神谷 信夫（大阪市立大学・複合先端研究機構・副機構長／複合先端研究機構・教授）
重川 直輝（大阪市立大学・複合先端研究機構・副機構長／工学研究科電子情報系専攻・教授）
天尾豊（大阪市立大学・人工光合成研究センター・所長／複合先端研究機構・教授）

吉田 朋子（大阪市立大学・人工光合成研究センター・副所長／複合先端研究機構・教授）
藤井 律子（大阪市立大学・人工光合成研究センター・分析装置運営部会会長／複合先端研究機構・准教授）
川上 恵典（大阪市立大学・複合先端研究機構・特任准教授）
野地 智康（大阪市立大学・複合先端研究機構・特任講師）
池山 秀作（大阪市立大学・複合先端研究機構・特任助教）
山口 良弘（大阪市立大学・複合先端研究機構・テニュアトラック特任准教授）
招待講演者
Dr. 銭 朴（シェフィールド大学、イギリス）
Dr. イリエシュ

ラウレアン（東京大学、日本）

Dr. Xing Yi Ling（南洋理工大学、シンガポール）
Dr. 菅原桂（株式会社 ジャパンティッシュ・エンジニアリング・再生医療事業（軟骨領域）首席、自家培養軟骨ジャック プロダクトマネージャー）

Dr. 寺川光洋（慶應大学・理工学部）

Dr. 浜口祐（理化学研究所・研究員）

Dr. 小澤

真一郎（岡山大学異分野基礎科学研究所・特任助教）

Dr. 上田昌宏（大阪大学 生命機能研究科・教授）

プロジェクトチーム講演者
中村 太郎（大阪市立大学・理学研究科生物地球系専攻・教授）
佐伯 壮一（大阪市立大学・工学研究科機械物理系専攻・准教授）
東海林 竜也（大阪市立大学・理学研究科物質分子系専攻・講師）
中曽 康壽（大阪市立大学・複合先端研究機構・特別研究員）
中尾 正喜（大阪市立大学・複合先端研究機構・特命教授）
吉田 朋子（大阪市立大学・複合先端研究機構・教授）
宮田 真人（大阪市立大学・理学研究科生物地球系専攻・教授）
寺北 明久（大阪市立大学・理学研究科生物地球系専攻・教授）
佐藤 哲也（大阪市立大学・理学研究科物質分子系専攻・教授）
重川 直輝（大阪市立大学・複合先端研究機構・副機構長／ 工学研究科電子情報系専攻・教授）

4Update: 20180213

The 9th OCARINA International Symposium

Joint mini-symposiums on Structural Biology and Catalysis

At 10th Floor, Media Center, Osaka City University, Osaka, Japan
March 6 (Tue)09:00-09:30 Registration
09:30-09:40 Opening 荒川 哲男（大阪市立大学・学長兼理事長）
Tetsuo ARAKAWA (President of OCU)
Session 1: バイオリソース(Bio-Resource)
09:40-10:00 中村 太郎 (理学研究科生物地球系専攻・教授)[日本語講演]
O1
ナショナルバイオリソースプロジェクト酵母
Taro NAKAMURA (Grad. School of Science, OCU)[Japanese lecture]
National BioResource Project yeast
10:00-10:10 BreakSession 2: バイオメディカル先端医療工学(Frontier Bio-Medical Engineering)
10:10-10:30 佐伯 壮一（工学研究科機械物理系専攻・准教授）[日本語講演]
O2
多機能 OCT を用いたマイクロ断層診断法～コスメおよび医療への応用～
Souichi SAEKI (Grad. School of Engineering, OCU) [Japanese lecture]
Micro-tomographic Diagnosis of Cosmetic and Medical Application using Multi-functional
OCT
10:30-11:20 招待講演 1（座長：佐伯 壮一 准教授）[日本語講演]
I1
菅原 桂（株式会社 ジャパンティッシュ・エンジニアリング・再生医療事業（軟骨領域）首席、自家培養軟骨ジャック プロダクトマネージャー）
自家培養軟骨ジャックの開発と製品化
Invited Lecture 1 (Chair: Souichi SAEKI) [Japanese lecture]
Katsura SUGAWARA (Japan Tissue Engineering Co. Ltd.)
Research, Development and Commercialization of an Autologous Cultured Cartilage,
JACC
11:20-11:30 BreakSession 3: ナノマテリアル光制御(Optical Control of Nano-Material)
11:30-12:20 招待講演 2（座長：坪井 泰之 教授）
I2
寺川 光洋（慶応義塾大学理工学部・准教授）[日本語講演]
フェムト秒レーザによる金属とソフトマテリアルの複合微細構造の作製
Invited Lecture 2 (Chair: Yasuyuki TSUBOI)
Mitsuhiro TERAKAWA (Keio Univ.) [Japanese lecture]
Fabrication of microstructures composed of metal and soft material by using femtosecond
laser
12:20-12:40 東海林 竜也（理学研究科物質分子系専攻・講師）[日本語講演]
O3
シリコンナノ構造を用いた NASSCA 光ピンセット法の開発
Tatsuya SHOJI (Grad. School of Science, OCU) [Japanese lecture]
Nano-structured semi-conductor-assisted (NASSCA) optical tweezers with black silicon
12:40-13:40
Lunch BreakSession 4: 都市エネルギー・防災(Urban energy)
13:40-14:10 中曽 康壽（複合先端研究機構・特別研究員）[日本語講演]
O4
高性能井戸を用いた帯水層蓄熱システムの開発と今後の普及展開
Yasuhiro NAKASO (OCARINA, OCU) [Japanese lecture]
Development of aquifer thermal energy storage system with high performance thermal welland future spread measures
14:10-14:40 中尾 正喜（複合先端研究機構・特命教授）[日本語講演]
O5
帯水層蓄熱空調システムの省エネルギー性能
Masaki NAKAO (OCARINA, OCU) [Japanese lecture]
Energy saving performance of developed aquifer thermal energy storage system
14:40-14:50 Break
Poster Session [Presentations in English]
14:50-16:20 Short Presentations [English presentations]
ポスター発表者によるショートプレゼンテーション[英語講演]
16:20-16:30 Break5

6

Update: 20180213
15:20-15:50
O10

Naoteru SHIGEKAWA (Grad. School of Engineering, OCU) [English lecture]
Low-temperature direct bonding of dissimilar materials for advanced electronics
重川 直輝（工学研究科電子情報系専攻・教授）[英語講演]
次世代エレクトロニクスを拓く低温直接接合
15:50-16:00 BreakMini Symposium on Catalysis（触媒ミニシンポジウム*注）
最寄り駅からの経路
16:00-16:50
Special Lecture 2 (Chair: Tetsuya SATOH)
S2Laurean ILIES (The Univ. of Tokyo) [English lecture]
C-H Activation catalyzed by earth abundant metals
特別講演 2（座長：佐藤 哲也 教授）
イリエシュ ラウレアン（東京大学理学研究科・准教授) [英語講演]
地球上に豊富に存在する鉄を触媒として用いる C-H 活性化
16:50-17:00 Break
17:00-17:50 Special Lecture 3 (Chair: Yusuke YAMADA)
S3
Xing Yi LING (Nanyang Technological University, Singapore) [English lecture]
Exploiting the nanoscale interface at metal nanoparticle and metal-organic framework forcatalysis and sensing applications
特別講演 3（座長：山田 裕介 教授）
Xing Yi LING（南洋理工大学・准教授）[英語講演]
金属ナノ粒子と金属有機構造体におけるナノスケールインターフェースの触媒およびセンシングへの応用
17:50-18:00
Break
18:00-18:10 Closing Nobuo KAMIYA (OCARINA, OCU, Vice Director of OCARINA)
神谷 信夫（大阪市立大学複合先端研究機構・副機構長）
18:10-18:20 Symposium Photo at 10F (記念撮影)
18:30-20:30 Banquet & Celebration for Poster Prize (1F Nonohana House)
JR 阪和線「杉本町（大阪市立大学前）駅」下車、東口徒歩５分
懇親会&ポスター賞授与式 (1F 野のはなハウス）*注

15 分
後援：平成地下鉄御堂筋線「あびこ駅」下車、４号出口より南西へ徒歩約
29 年度大阪市立大学教育推進本部経費事業「研究科横断型大学院教育改革の推進と化学人材育成」

喫煙エリア

7

Guidelines for Poster Presentations（1）Preparation for the poster◦ The poster board provided is approximately 160 cm height x 110 cm width.
◦ Please limit the size of your entire poster or poster sections in it. (Recommended poster size is A0.)
◦ Please place your poster on the poster board numbered for you.
◦ Push pin will be available at the site.
◦ Poster viewing will be start from odd number, and then even number (40 min each). During your time,please stay around the poster.
◦ Display your poster during OCARINA symposium (6 March, 9:00 - 7 March, 18:00).
（2）Preparation for the short oral presentation◦ All the poster presenters can present own short oral presentation.
◦ One separate file (pdf or power point format) is available to show during your presentation.
◦ No animation. Submit the file to the conference secretary.
◦ Gather and make a queue in front of the stage before starting of the short oral presentation.
Allocated time is 1 minute including set up time. Please stand up nearby the microphone and be
ready before finishing of the previous talk.

ポスター発表について（１）添付ポスターについて（英語）◦ポスターサイズ

A0（ボードサイズは W110 x H160 cm 程度 )◦添付：自分のポスター番号（要旨集／ HP）の場所に添付してください。

◦会

場：学術情報センター 10F

研究者交流室内◦添付期間：3 月 6 日（火）9 時 〜 3 月 7 日（水）18 時頃まで
◦ポスター賞の審査を希望する方は、必ず最後まで貼付してください。
◦ポスターの撤去は、ご自身でお願いします。放置されたポスターについてはこちらで処分いたします。
◦ポスター発表：前半／奇数番号

後半／偶数番号➢ 自分の発表の時間帯は、自身のポスターのそばにいてください。
（２）ポスターショートプレゼンテーションについて（英語、持ち時間１分（厳守））
上述赤字の注意事項を熟読のこと。
◦ポスター賞審査方法

招待講演者及び学内複合先端研究機構プロジェクトメンバー有志の審査員による審査。異分野の研究者に自身の研究を的確に伝えているかどうか。
（研究の背景・目的・意義・狙いを理解してもらい、討論をする。
）◦ポスター賞授与式 3 月 7 日（水）18 時 30 分開始の懇親会場にて行います。皆様どうぞご参加
ください。

8

Oral Presentations
March 6th (Tue), 2018

National BioResource Project yeast
Taro Nakamura
Graduate School of Science, Advanced Research Institute for Natural Scienceand Technology, Osaka City University, Sugimoto 3-3-138, Sumiyoshi-ku, Osaka
The National BioResource Project (NBRP) starts in FY 2002 to comprehensively promote life
sciences. In the project, the systems of collection, preservation, and deposition have been
established for bioresources, such as experimental animals, plants, and microbes (systems,groups, tissues, cells, and genetic materials of animals, plants, and microbes and theirinformation as research and development materials) that are important for the nation toorganize strategically.
Yeast is an important eukaryotic model organism. This is especially true of thefission yeast Schizosaccharomyces pombe and the budding yeast Saccharomyces cerevisiae,
which are making significant contributions to research in a variety of areas within the life
sciences. NBRP-Yeast has established a framework to collect, stock and distribute strain and
DNA resources of mainly the two species. Through phases 1 and 3 (FY 2002 - 2016) of the
NBRP, the NBRP-yeast has become one of the top international yeast resource centers. Here,I will introduce the activity of NBRP-yeast.

O-1

11

Micro-tomographic Diagnosis of Skin and Regenerative Tissue
using Multi-functional OCT
Souichi SAEKI1
1

Graduate School of Engineering, Mechanical & Physical Engineering, Osaka City
University. 3-3-138, Sugimoto, Sumiyoshi-ku, Osaka, 558-8585

Abstract:
Water has significant influence on human body due to physiological carrier media. In addition,rheological behavior of interstitial fluid in epidermal and dermal tissue, including blood microcirculation, can vary skin mechanics in micro scale, i.e. vis. Therefore, an in vivo measurementof moisture content and capillary blood flow velocity is necessary to clarify skin mechanics.
This paper presents 2C-OCM (2-Color Optical Coherence Moisturegraphy), which wascomposed of two-band light sources having different optical absorption properties of water
respectively. These are capable of tomographically in vivo diagnosing the moisture content.
Furthermore, OCDV (Optical Coherence Doppler Velocigraphy) algorithm can visualize thetomographic flow velocity of red blood cell in capillaries of human epidermal skin. In thisexperiment, 2C-OCM & OCDV were in vivo applied to human skin and regenerative tissue.
Consequently, it was concluded that 2C-OCM & OCDV can provide micro tomography ofmoisture content and capillary blood velocity inside skin tissue, which can be attributed tobiomechanical properties.

O-2

References:[1] Daisuke Furukawa, et al., “Accuracy Evaluation on Tomographic Micro-visualization of
Flow Velocity using High Frequency Modulated Low Coherence Interferometer, J. JSEM,
Vol.17, Issue 1, (2017), pp.52-56.
[2] Yusuke Hara, Toyonobu Yamashita, Kumiko Kikuchi, Yoshihide Kubo, Chika Katagiri,
Kentaro Kajiya, Souichi Saeki, “Visualization of age-related vascular alterations in facial skinusing optical coherence tomography-based angiography”, Journal of Dermatological Science,
2018, Jan. 8, S0923-1811(18).
[3] Yusuke Hara, Yuki Ogura, Toyonobu Yamashita, Daisuke Furukawa, Souichi Saeki,“Visualization of visco-elastic behavior in skin equivalent using optical coherence
tomography straingraphy”, Skin Research and Technology, 2018, Jan, 24, 10.1111/srt.12435.

12

Research, Development and Commercialization of
an Autologous Cultured Cartilage, JACC
Katsura SUGAWARA
Japan Tissue Engineering Co., Ltd., Gamagori, Aichi, Japan
Abstract: Recently, regenerative medicine has made remarkable progress and attracted
attention worldwide. Articulat cartilage is a thin layer that covers the bones in knee joints andallowing them to move smoothly. However, since articular cartilage lacks blood vessels andnerves, once cartilage is damaged by trauma or overuse, it is not able to heal spontaneously.
Brittberg and Peterson et al. performed autologous chondrocyte implantation (ACI) in
patients with full- thickness cartilage defects in the knee [1]. In this procedure, cartilage
pieces are taken from the unloaded resion of the knee and isolated chondrocytes are cultivatedfor two to three weeks. After the cultivation, chondrocytes are harvested and implanted into
the cartilage defect of the patient in a cell suspension manner with a periosteum patch. This
regenerative medicine for the cartilage repair was an epoc-making invention, however, somedisadvantages were pointed out; 1) during the cultivation, chondrocytes lose their ECM
producing activity 2) possible leakage of chondrocyte after implantation due to its formulation
3) an uneven distribution of chondrocytes because of gravity. To address these issues, Prof.
Ochi at Hiroshima University developed three-dimensional cultivation of chondrocytes [2]. In
the procedure, chondrocytes are embedded in atelocollagen gel and cultivated for three to fourweeks. On the basis of basic and animal studies, clinical studies were conducted since 1996 at
Shimane Medical University and Hiroshima University. This technology was transferrd to
Japan Tissue Engineering Co., Ltd. (J-TEC) and a multi-center clinical trial was conducted in
2004 [3]. J-TEC obtained the approval for the three-dimensional cultured cartilage, named
JACC, in 2012 as the first orthopaedic regenerative medicinal product in Japan and JACC
was listed as an item covered by the National Health Insurance from 2013.
In this presentation, translational research, development and commercialization of JACC will
be reviewed and discussed.

I-1

References:[1] Brittberg, M.; Lindahl, A.; Peterson, L. et al., New Engl J Med 1994, 331, 889.
[2] Ochi, M.; Uchio, Y.; Kawasaki, K. et al., J Bone Joint Surg 2002, 84, 571.
[3] Tohyama, H.; Yasuda, K.; Ochi, M. et al., J Orthop Sci 2009, 14, 579.

13

I-2

14

Nano-structured semi-conductor-assisted (NASSCA) opticaltweezers with black silicon
Tatsuya Shoji
Graduate School of Science, Osaka City University
Abstract: We demonstrated a novel optical manipulation technique for nanoparticles with
non-plasmonic semiconductor nanostructures; named nano-structured semi-conductor-assisted(NASSCA) optical tweezers[1]. A tightly focused laser beam exerts optical force on polarized
microparitcles such as living cells, bacteria, and liquid droplets, leading to optical trappingand manipulation of them (optical tweezers). However, optical manipulation of smaller sized
nanoparticles and molecules is a still challenging issue because of their quite small
polarizability.
Instead of conventional optical trapping methods, we present an alternative and novel
trapping method – NASCA optical tweezers – using metal-free black silicon (B-Si) whose
surface has nano-needle structures. NASSCA optical tweezers have several advantages: (1)simple fabrication with high uniformity over wafer-sized areas (cm×cm), (2) free fromthermal effects detrimental for trapping, (3) switchable trapping between one andtwo-dimensions, (4) tight trapping because of no detrimental thermal forces, and so on. We
believe that NASSCA optical tweezers will become an indispensable tool for manipulating
nanoparticles and molecules in the same way that conventional optical tweezers have been
proven to be a valuable micromanipulation tool.

O-3

References:[1] Shoji, T.; Mototsuji, A.; Balčytis, A.; Linklater, D.; Juodkazis, S.; Tsuboi, Y. Sci. Rep.
2017, 7, 12298.

15

Development of Aquifer Thermal Energy Storage System withHigh Performance Thermal Well and future spread measures
Yasuhisa Nakaso
Reaserch Fellow, 3-9-11,Sugimoto,Sumiyoshi-ku,Osaka City, Japan
Abstract:
Large urban areas in high energy consuming density are required to applicable practicalscale renewable energy technologies without damaging urban environment like global warmingand heat island problem. In some Japanese urban area, renewable thermal energy utilization of
sewage and river water has been realized as district heat and cool supply plant.
Now, we can realize direct groundwater heat source utilization under iron and salt ion
rich location in the coastal urban area, that leads hard well clogging easily and so on which wasdifficult to use, and this technical development project is assisted by Japanese Ministry of theEnvironment.
We set up the well in the Umekita redevelopment area in the northern part of JR Osaka
Station, and we tried pumping up a groundwater 100m3 per hour from a well, getting a heat
power and immediately rechrging the groundwater from another well to original same aquifer.
This is better way that reduce the influence of land subsidence problem in the urban areawith keeping groundwater level in local area, and we confirmed it could be operated stably for along period by using airtight structure.
In this paper, we introduce the background and the outline of this technology
development including the preceding trial in Takasago area.

O-4

Osaka Umekita site
Sampling tube↓#2 well#1well&Thermal equipment

See through pipe

#1well (cold well)

試験実施個所の概観

Propaganda in Umekita site.

#1well connection←After1H itturns
brown.

・Radiation panel through a ground water, fell cool insummer.
Cut modell of well

Site tour

16

1/200 model

After pumping up, groundwater turns
white. (including methane)

Boring sample
before 20,000 years←It is no
problemfordrinking,
but badtaste.

Measurement
bar through thefirst aquifer

Criteria of drawdown vs. abstraction and injection volume
Q

rw

Abstraction volume Q, Permeability k, Drawdown hw

Rr
Aquitard

Aquiferhwmi=dh/drh→They have proportional relationh0

permeabilityｋ

Aquitard

Clogging occurred
near bore wall whichhave high watervelocitySame water vlume brings
large drawdown

Takasago site operation report

Flow rate

Draw down

Draw down vs. flow rate
10/31

10/31

10/31

10/31

2016/10/31

2016/11/26

2017/1/24

2017/2/24

O-4
10/31

10/31

10/31

10/31

2017/4/10

2017/6/16

2017/8/18

2017/12/08

エネルギー消費密度の高い都市域において、地球温暖化防止やヒートアイランド問題など都市環境を損なうことなく、適用可能な実用規模の再生可能エネルギー技術の普及開発が求められている。
わが国においても、地域熱供給事業として下水や河川水の熱利用が実用化されている。
この度、環境省の技術開発実証事業として始めた地下水の熱源利用研究において、臨海都市部
で鉄分や塩分濃度が高く、これまで目詰まりを起こすなどして利用の難しかった熱源井の開発に成功した。
この井戸は JR 大阪駅北のうめきた地区の再開発エリアの一角に地下水を汲み上げるための井戸を設置し、実用規模レベルである毎時 100m3 で揚水を行い、熱利用後、直ちに元の帯水層に還元することで、地域の地下水レベルの低下を防止、都市域で問題となっている地盤沈下の影響を軽減すると共に、気密構造の徹底により、長期間、安定運用できることも確認した。本稿では、先行する高砂地区での事例も含め、この技術開発の背景と取り組み概要について紹介する。

17

Energy Saving Performance of Developed Aquifer Thermal
Energy Storage System
Masaki Nakao
Professor, 3-9-11,Sugimoto,Sumiyoshi-ku,Osaka City, Japan
Abstract:
Aquifer Thermal Energy Storage System(ATES) is storage and recovery of thermalenergy system which is achieved by extraction and injection of groundwater from aquifers
using two groundwater wells.
ATES is a technology developed in the Netherlands, and it has already been installedin more than 1,000 facilities in the country.
In order to apply it to our country it is necessary to operate and control the thermal
storage facility in consideration of the Japanese climate conditions, the cooling and heatingload characteristics of the building, and the regional level of the aquifer temperature.
In this report, we show the test result and energy saving effect by introducing the
ATES to the heating and cooling of the factory located in the coastal area. In addition, theverification results of the simulation model linking the aquifer and heating and coolingequipment is described. Furthermore, we show the effect of energy conservation when
applying ATES to an office building in Osaka City using simulation model. In particular, it
shows that it is effective to cool the aquifer of the low temperature well at low outside
temperature in the winter season.

O-5

In order not to affect adjacent ATES, it is necessary to handle the heat transfer
performance as a closed system. Further, it is necessary for the aquifer storage to perform
stably for a long term. So, the following operation of ATES is discussed for long-term
operation.
1) Prevent long-term aquifer temperature rise or decline.
2) The heat mass size of each well stabilizes
As a countermeasure, the spacial average temperature of the aquifer does not changeover time. Moreover, the annual cumulative extraction of each well is equal to injection ofgroundwater amount.
In order to satisfy these two conditions, we devise an operation method of the ATES
that changes the operation parameters of the following year from the results of the previous
year.

18

Intermediate
periodWinterStorage of chilled water isachieved by the operationof a heatpump

Storage of chilled water isachieved by outdoor air
coolingHeating load

Heat
pump

Heat
pump

Intermediate
periodSummerStorage of hot water isachieved by a heatpump

Cooling
tower

Cooling load

Cooling
tower

Heat
exchanger

Heat
pump

Heat
exchanger

Annual cycle of ATES operation
Heating
43℃

850kW

6℃

O-5

Fancoil
空調機unit

Turbo-Heatpump

Well

Chilled water storage

Actual plant of heating and cooling system for the Factory in Takasago City
Actual operation of heating
and cooling system for the
Factory in Takasago City
Factoryplant

Experimental
verification of model
Modeling of heating andcooling plant
Interconnected simulation
methodEstimation of energy saving effectwhen ATES is applied to a Central
district office building in Osaka

Office building

Interconnected simulation

Identification method ofthermal storage
characteristics of aquifer
Aquifers using two groundwater
wells in Coastal Area

Identified model of aquifer inCentral area in Osaka

Modeling, experimental verification and estimation of energy saving effect of ATES

19

20

Oral Presentations
March 7th (Wed), 2018

Visualization and structures of membrane proteinsby cryo-electron microscopy
Tasuku Hamaguchi, Saori Maki-Yonekura, Koji Yonekura
RIKEN SPring-8 center
Structures of life’s molecular machinery are indispensable to understand biologicalphenomena in details. The current technology, however, still suffer from limitations such as
crystallization, stability, heterogeneity and so on. Cryo-electron microscopy (cryo-EM)
allows to obtain the 3D structures of biological macro molecules under a natural condition ora condition close to it without crystallization, heavy-atom staining and chemical fixing.
Cryo-EM has determined the structures of membrane proteins and protein complexes, e.g.
GPCRs, ion channels, transporters, and rotary-ATPases, and the number of high-resolution
structures has been increasing explosively, since the direct electron-detection camera wasintroduced in the field and the structure of TRPV1 was reported at a near-atomic resolution
using this new technology [1]. Furthermore, cryo-EM can visualize super-molecularmachineries, which are hard or impossible to be crystalized. Those targets include bacterial
flagella, injectosome, and cell organelles.
Exosome, an extracellular vesicle excreted from eukaryotic cells, plays an importantrole in intercellular communications. Reflecting dispositions of the derived cells, theexosome contains many soluble proteins, micro-RNAs and membrane proteins. The
exosome-specificmembraneproteins
includeexcitatory
amino-acidtransporters
andlysosome-associated membrane glycoproteins. The
former regulates an excitation of the postsynaptic
action, and this transporter expressed in a central
nervous system is responsible for various stresses.
We aim at visualization of these targets by cryo-EM.
In addition, a membrane protein machinery involved inbacterial motility is also analyzed by cryo-EM and will
be presented in this workshop.

I-3

Cartoon of exosome[1] Liao, M., Cao, E., Julius, D., and Cheng, Y. Nature 2013
504(7478), 10723

Cryo-EM structure of the Blastochloris viridis
RC-LH1 complex at 2.87 Å
Pu QianDepartment of Molecular Biology and Biotechnology, University of
Sheffield, Sheffield, UK.
The reaction centre light-harvesting 1 (RC-LH1) complex is the core functional component ofbacterial photosynthesis. A 2.87 Å resolution cryo-EM structure of the bacteriochlorophyll bbased RC-LH1 from Blastochloris viridis reveals the structural basis for absorption of
infrared light, and the molecular mechanism of quinone migration across the LH1 complex.
The novel triple ring LH1 complex comprises a circular array of 17 β-polypeptides
sandwiched between 17 α- and 16 γ-polypeptides. Tight packing of the γ-apoproteins between
βs collectively interlocks and stabilizes the LH1 structure which, together with the short MgMg distances of BChl b pairs, contributes to the large red-shift of bacteriochlorophyll babsorption. The ‘missing’ 17th γ polypeptide creates a pore in the LH1 ring, and an adjacentbinding pocket provides a folding template for a novel quinone, QP, which adopts a compact,
export-ready conformation prior to passage through the pore and eventual diffusion to thecytochrome bc1 complex.

S-1

24

Configuration of Antenna subunits in Photosystem I complex
Shin-Ichiro Ozawa
Research Institute for Interdisciplinary Science, Okayama University,
Okayama, Japan
Abstract: In oxygenic photosynthesis, electron transfer is driven by two types ofphotosystems, Photosystem II (PSII) and Photosystem I (PSI), operating in tandem. In
plants and algae, the association of antenna complex, light-harvesting chlorophyll a/b
complex (LHC), significantly increases light-harvesting capacity. LHC consists of LHC
subunits of which structures, as well as chlorophyll binding sites, are well conserved.
However, the physical property of individual LHC subunit is not identical because chemical
species and arrangement of chlorophyll are slightly modified. PSI binds LHCI while PSII
binds LHCII, respectively and specifically, to form PSI-LHCI or PSII-LHCII, and it was
found that configurations of each LHC subunit are well organized. Structure of PSI-LHCI
from pea is resolved at 2.6 Å by X-ray crystal structural analysis and individual LHCI
subunit configuration is well described [1]. According to the crystal structure, four LHCI
subunits (Lhca1-4) are located in a half-ring arrangement. In contrast to the plant, thearrangement and stoichiometry of LHCI subunits in algal PSI are not sufficiently
determined because of the difficulties to isolate intact PSI-LHCI complexes and/or thepresence of more LHCI subunits.
It is currently accepted that PSI-LHCI supercomplex of Chlamydomonas reinhardtii has
nine distinct LHCI subunits (LHCA1-9). Our knowledge of the configuration of individual
LHCI subunits is even more limited while the most recent single particle analysis suggested
that nine LHCI subunits are arranged in two layers [2]. Recently, our group determinedboth the stoichiometry and configuration of nine distinct LHCI subunits in PSI-LHCI
supercomplex of C. reinhardtii[3]. To determine the stoichiometry of the LHCI subunits,
we employed uniform labeling of total cellular proteins with 14C, subsequently, theconfiguration of all LHCI subunits in PSI-LHCI supercomplex was determined by chemical
cross-linking in combination with the identification of cross-linked products byimmunoblotting and mass spectrometry.
In this talk, I am going to introduce LHCI briefly and discuss functional insights of LHCI
subunits in C. reinhardtii.

I-4

References:[1] Mazor, Y.; Borovikova, A.; Caspy, I.; Nelson, N., Structure of the plant photosystem I
supercomplex at 2.6 Å resolution. Nat Plants 2017, 3, 17014.
[2] Drop, B.; Webber-Birungi, M.; Fusetti, F.; Kouril, R.; Redding, K. E.; Boekema, E. J.;
Croce, R., Photosystem I of Chlamydomonas reinhardtii contains nine light-harvesting
complexes (Lhca) located on one side of the core. J Biol Chem 2011, 286 (52), 44878-87.
[3] Ozawa, SI.; Bald, T.; Onishi, T.; Xue, H.; Matsumura, T.; Kubo, R.; Takahashi, H.;
Hippler, M.; Takahashi, Y., submitted.

25

XAFS/EELS analyses of nitrogen doped titanium oxide
photocatalysts
Tomoko Yoshida1, Muenaki Yamamoto2, Akiyo Ozawa3,
Yuma Kato3, Shinya Yagi4, Tetsuo Tanabe1
1Advanced Research Institute for Natural Science and Technology, Osaka City University
2
Materials, Physics and Energy Engineering, Graduate School of Engineering, Nagoya
University
3
Applied Chemistry and Bioengineering, Graduate School of Engineering, Osaka City
University
4
Institute for Materials and systems for sustainability, Nagoya University

Photocatalytic reactions at the surface of titanium dioxide (TiO2) under UV lightirradiation have been attracting much attention in view of their practical applications toenvironmental cleaning such as self cleaning of tiles, glasses, and windows. Recently, it was
found that the doping of nitrogen into TiO2 contributed to narrowing of the band gap, thus
providing a visible-light response [1]. Previous studies demonstrated that nitrogen doping
generated new optical absorption bands in the visible-light region and the absorbance evolved
with increasing nitrogen. On the other hand, absorbance may not be linearly proportional tothe photocatalytic activity [2], and it is thus important to understand the chemical state of
doped nitrogen most effective for the visible-light response.
The N K-edge XANES spectrum of the photocatalytic active nitrogen doped TiO2
showed a characteristic double-peak at 398 and 401 eV, and the XANES spectrum of theinactive photocatalyst a distinct single peak around 401 eV. These features of the XANES
spectra were well reproduced by theoretical simulations based on the model where an O atom
in TiO2 were replaced by N ((N)s) for the active photocatalyst, and that of NO2 ((NO2)s) for
the inactive photocatalyst [3]. The (N)s had proven to be indispensable for band-gap
narrowing and photocatalytic activity, and the optimal nitrogen concentration for generating
the (N)s was evaluated to be less than 1.8 atom% by EELS analysis.
We also tried to visualize these two different nitrogen species in photocatalytic active andinactive nitrogen doped TiO2 samples by introducing modern mathematical treatments toenergy-filtering TEM. This advanced nanosized mapping clearly showed that the spatial
distributions of both (N)s and (NO2)s species in TiO2 are the important key to generate
visible-light response of a TiO2 photocatalyst.

O-6[1] Asahi R., Owaki T., Aoki K., Taga Y., Science. 2001, 293, 269-267[2] Nosaka Y., Matsushita M., Nishino J., Nosaka A. Y., Sci. Tech. Adv. Mater., 2005, 6,
143-148.
[3] Yoshida T., Niimi S., Yamamoto M., Ogawa S., Nomoto T., Yagi S., Nucl. Instr. andMeth. B, 2015, 365, 79-81

26

Recommendation for quick freeze replica
electron microscopy
Makoto Miyata
1) The OCU Advanced Research Institute for Natural Science andTechnology2) Graduate School of Science, Osaka City University
Quick freeze replica electron microscopy was developed to visualize membrane fusion at asynapse of nerve cell in 1979, and mainly applied to animal cell physiology. In this method,a specimen is frozen and fixed in a moment by slamming it onto a cupper block cooled byliquid helium or nitrogen, and visualized three dimentionally as a pratinum replica. It gives
high image contrast, nanometer spacial and submillisecond time resolutions, which cannot be
achieved by any other methods. However, recently only a few groups can provide thismethod over the world, because it needed “advanced craftmanship”. In other words, thisexciting special method was going before our project.
Our project is to change this method more friendly to new researchers and subjects. We
welcome both intra and inter university collaborations on various subjects including (i)motility machinery, (ii) pathogenic microorganisms, (iii) usefulmicroorganism, and (iv) materials. I will introduce exciting images
recently obtained in our project.

O-7

References[1] Heuser, J.E. J Electron Microscopy (Tokyo). 2011, 60 Suppl 1:S3-29.
[2] Trussart, M.; Yus, E.; Martinez, S.; Baù, D. Tahara, Y.O.; Pengo, T.; Widjaja, M.;
Kretschmer, S.; Swoger, J.; Djordjevic, S.; Turnbull, L.; Whitchurch, C.; Miyata, M.;
Marti-Renom, M.A.; Lluch-Senar, M.; Serrano, L. Nature Communications. 2017, 8, 14665

27

Molecular physiology of diverse light-senor proteins and their
optogenetic applications
Akihisa Terakita and Mitsumasa Koyanagi
Department of Biology and Geoscience, Graduate School of Science, Osaka City
University, Osaka, Japan; OCARINA, Osaka City University, Osaka, Japan

Most animal opsins, which bind to a chromophore retinal to form photosensitive pigments(opsin-based pigments),
serve as light-sensitive G protein-coupled receptors (GPCRs),
which constitute a large protein family of receptors that sense molecules outside the cell andactivate inside signal transduction cascades and, ultimately, generate cellular responses.
Many animals capture light information through opsin-based pigments and utilize theinformation for visual and non-visual functions including regulation of circadian rhythms.
Thousands of opsins have been identified from a wide variety of animas thus far. We havecharacterized diverse opsins, including novel ones, spectroscopically, biochemically,molecular physiologically and evolutionally [1-4], and also investigated their contribution tobiological function [5]. Interestingly, we found that some non-visual opsins have uniquemolecular properties suitable for optogenetics applications, which is the combination ofgenetics and optics to control well-defined events within specific cells of living tissues and is
now a strong application to control cell activities and animal behaviors by light.
A pineal UV-sensitive opsin parapinopsin, which is activated by UV light and deactivated bygreen light, contributes to the pineal function of lower vertebrate, discrimination of
wavelength of lights, UV and green lights [1, 4]. We found that UV and green lightilluminations activate and deactivate G protein-mediated signal transduction cascade in the
mammalian cultured cells expressing parapinopsin [6]. Therefore we suggest thatparapinopsin has optogenetic potentials to regulates cellular responses and animal behaviours
in a light wavelength-dependent manner.

O-8

References:[1] Koyanagi et al., Proc. Natl. Acad. Sci. U.S.A 2004, 101, 6687
[2] Tsukamoto et al., J. Biol. Chem 2009, 284, 20676[3] Koyanagi et al., Proc. Natl. Acad. Sci. U.S.A 2013, 110, 4998
[4] Koyanagi et al., BMC Biology 2015, 17, 73[5] Nagata et al., Science 2012, 355, 469[6] Kawano-Yamashita et al., PLoS One 2015, 10, e0141280

28

Automated single-molecule imaging analysis of cell signaling
Masahiro Ueda
Graduate School of Frontier Biosciences, Osaka University
and Quantitative Biology Center (QBiC), RIKEN
Abstract:
Single-molecule imaging of biomolecules in living cells allows for the investigation of cellsignaling and other molecular mechanisms [1]. The techniques have made it possible todirectly monitor the behaviors of biomolecules in living cells, in which the locations,movements, turnovers, and complex formations of biomolecules can be detected
quantitatively with single molecule sensitivity, providing powerful tools to elucidate
molecular mechanisms of intracellular signaling processes. The systematic andcomprehensive measurement of a huge number of molecular species at single moleculesensitivity provides detailed information on elementary biological processes and new insights
into a system’s dynamics, deepening and extending the current biological and medical
knowledge. However, technical expertise has been required for both microscope operationand data analysis, which has prevented the analysis from being a standard in medical andbiological research. Recently, we have developed an automated single-molecule imaging
apparatus for live-cell analysis [2]. All the significant procedures, such as searching for cells
suitable for the observation, detecting an in-focus position, image acquisition andsingle-molecule tracking, are fully automated, allowing us to accomplish a large amount ofhighly accurate, efficient, and reproducible live-cell imaging experiments. The apparatus
could complete single-molecule imaging and analysis of epidermal growth factor receptor(EGFR) for 1,600 cells in a 96-well plate within one day. Changes in the lateral mobility of
EGFR on the plasma membrane in response to various ligand and drug concentrations weredetected clearly in individual cells, which provided several dynamics and pharmacologicalparameters including diffusion coefficient, oligomer size, and EC50. Our data show that upon
EGF stimulation, EGFR gradually undergoes a transition from the fast-mobile state to slow
and immobile states within a 270 nm diameter wide area and simultaneously forms large
oligomers that function possibly as a signaling hub for the downstream signaling molecule
Grb2. Automated single-molecule imaging is feasible for the systematic analysis of cellsignaling and can also be applied to single-molecule screening, thus extensively contributing
to biological and pharmacological research.

I-5

References:[1] Ueda, M. et al. Science 2001, 294, 864.
[2] Yasui, M.; Hiroshima, M.; Kozuka, J.; Sako, Y; Ueda, M., under revsion.

29

Synthesis of -Conjugated Molecules
via Transition-Metal-Catalyzed C–H Functionalization
Tetsuya Satoh
Department of Chemistry, Graduate School of Science, Osaka City University,
3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan
Transition-metal-catalyzed regioselective C–H functionalization reactions with the aid of
directing groups (–DG) are recognized as important tools for environmentally benign
synthesis, since they provide atom- and step-economical routes toward complex moleculesfrom simple substrates. Especially, oxidative coupling of aromatic C–H and unsaturated
compounds in the presence of an appropriate oxidant are highly useful for constructing
π-conjugated molecules. In earlier examples for such direct transformation, palladiumcatalysts used to be employed. However, homogeneous palladium catalysts tend to be easily
inactivated under oxidative conditions. In addition, substrate scope used to be relatively
narrow.
R
R

R

R
DG

H
DG

O-9

TM-cat.
oxidant
R

R
DG

We succeeded in finding that Cp*Rh(III) complexes are stable under oxidative conditions andcan effectively promote the oxidative coupling reactions [1]. Particlarly, the annulative
coupling of aromatic substrates possessing various heteroatom-containing directing groups
with internal alkynes proceeds smoothly under rhodium catalysis to give the corresponding
fused heteroaromatic molecules [2].
References:[1] For review, see: Satoh, T.; Miura, M. Chem. Eur. J. 2010, 16, 11212.
[2] For representative examples, see: (a) Morioka, R.; Nobushige, K.; Satoh, T.; Hirano, K.;
Miura, M. Org. Lett. 2015, 17, 3130. (b) Iitsuka, T.; Hirano, K.; Satoh, T.; Miura, M. J. Org.
Chem. 2015, 80, 2804.

30

Low-temperature direct bonding of dissimilar materials
for advanced electronics
Naoteru Shigekawa
Gradutate School of Engineering, Osaka City University
Seven research projects are currently going on in OCARINA. As members of “Frontier
Materials” Project, one of the seven research projects, we have been investigating applicationof surface-activated bonding (SAB), which is one of low-temperature direct bonding
technologies, for fabricating junctions of dissimilar materials. We previously fabricated
III-V-on-Si multijunction solar cells [1] and metal-foil-based ultrathick interconnects directly
bonded to semiconductor substrates [2] by using SAB. In this presentation, we focus on
possibility of SAB-based junctions for realizing novel high-power devices.
The performance of high-power devices is mainly limited by the deterioration in escape ofheat during the operation. Diamonds are promising as materials for thermal spreaders becauseof their excellent insulating properties and large thermal conductivity. It was reported that a
diamond film was directly bonded to a Si (100) substrate [3]. The bonding process, however,required a long-period (15 h) loading in a high temperature (> 950 C), which prevents themethod from being applied in semiconductor industries.
We successfully fabricated diamond/Si junctions by using SAB without heating samples
during bonding [4]. We found that diamond and Si were firmly bonded to each other by using
an SEM. An observation using a TEM and EELS indicated that both sp2 and sp3 bondings
were distributed in a diamond layer neighboring the bonding interface. We confirmed by XPS
characterization that the sp2 bondings were formed during irradiating fast atom beams of Ar
for activating surfaces of samples. It is assumed that the mixture of sp2 and sp3 bondings
formed on a surface of diamond should play an essential role in successfully bonding
diamond and Si at low temperatures.
The authors are grateful to Prof. K. Tsuji in Graduate School of Engineering, Osaka City Univ.
for his advice in XPS characterization. This work was supported by the Grant-in-Aid for
Challenging Exploratory Research (16K13676) of the Ministry of Education, Culture, Sports,
Science, and Technology (MEXT), Japan.

O-10[1] N. Shigekawa, et al. Jpn. J. Appl. Phys. 54, 08KE03 (2015).
[2] J. Liang, et al. ECS J. Solid State Sci. Technol., 6, 626 (2017).
[3] G. N. Yushin, et al. Appl. Phys. Lett. 81, 3275 (2002).
[4] J. Liang, et al. Appl. Phys. Lett. 110, 111603 (2017).

31

S-2

32

Exploiting the Nanoscale Interface at Metal Nanoparticle andMetal-Organic Framework for Catalysis and Sensing
Applications
Xing Yi Ling
Division of Chemistry and Biological Chemistry, School of Physical andMathematical Sciences, Nanyang Technological University, 21 Nanyang Link,
Singapore 637371.

E-mail: xyling@ntu.edu.sg

Abstract. A major challenge in nanoparticle self-assembly is programming the large-areaorganization of a single type of anisotropic nanoparticle into distinct superlattices with tunable
packing efficiencies. In this talk, I will discuss how nanoscale surface chemistry is used to direct
the self-assembly of shape-controlled silver nanoparticles into three distinct two-dimensional
plasmonic superlattices at a liquid/liquid interface. Systematically tuning the surface chemistry of
the silver nanoparticles leads to a continuous superlattice structural evolution, from close-packed
to progressively open structures. Notably, silver octahedra standing on vertices arranged in asquare lattice is observed using hydrophobic particles. The structure-to-function characterization
reveals that the nanoparticle assembly with the least packing density generates plasmonic‘hotstrips’, leading to nearly 10-fold more efficient surface-enhanced Raman scattering comparedwith the other more densely packed configurations.
In the second part, I will discuss how real-time SERS is developed to directly observe theconcentration of CO2 molecules into a quasi-condensed phase at a nanoscale interface at a
metal-organic framework (MOF) and metal nanoparticle surface, even at ambient conditions of 1
bar and 298 K. A systematic spectral evolution is observed throughout our time-dependent SERS
studies, which enables us to experimentally visualize the in situ linear-to-bent transformation of
CO2 at the solid-gas interface with increased gas accumulation over time. These direct
observations highlight the advantage of surface-sensitive SERS in probing such sophisticatedmolecular dynamics occurring at the enclosed surface of solid@MOF system, offering valuable
insights to further our understanding in solid-gas interactions at ambient operations.

S-3

33

34

P-1

Au-nanoparticle-embedded polymer films
synthesized on HAuCl4/gelatin aqueous solution
irradiated with Ar DBD plasma
1Junji Kambayashi, 1Shiori Azuma, 1Yusuke Nakamura,2
Toshiyuki Isshiki, and 1Tatsuru Shirafuji
1
Department of Physical Electronics and Informatics,
Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka, 558-8585 Japan2
Department of Electrical Engineering and Electronics,
Kyoto Institute of Technology, Kyoto 606-8585, Japan

2. Experimental procedure
The aqueous solution was HAuCl4 (0.15, 0.30, and 0.60mM) aqueous solution with gelatin (5, 10, and 20 wt.%). The
DBD system used in this study is shown in Fig. 1. Applied
voltage was bipolar pulse voltage (amplitude 4 kV, frequency
40 kHz, pulse width 4 μs). Typical discharge time was 10
min. Discharge gas was argon.

gas
inlet

100 mm

1. Introduction
Solution plasma can be used for various applications
including nanoparticle synthesis and liquid treatment [1].
However, there are few reports on thin film formation using
solution plasma [2]. In this work, we report that we cansynthesize a free-standing film, which is cross-linked gelatinwith embedded gold nanoparticles (GNPs), on an aqueous
solution irradiated with dielectric barrier discharge (DBD).

to HV

gas
outlettop
metalelectrodediameter
10 mm

top
dielectric
diameter
20 mm

This side
is made ofquartz plate
for OES.

top
dielectric
thickness
2 mm

gap
2 mm depth
8 mmbottomdielectric
thickness
2 mmbottommetalelectrodediameter
20 mm

OES
through
an opticalfiber

to GND

Fig. 1 The DBD reactor used inthis study.

3. Results and discussion
100nm
A film is formed on the aqueous solution by irradiating DBD
plasma on the aqueous solution. Infrared absorption spectra
Fig. 2 TEM image of the film.
of the film and energy dispersive X-ray spectra haveindicated that the film is made of cross-linked gelatin and gold. Figure 2 shows a transmission
electron microscope (TEM) image of the sample taken from the outer edge of the film, whichindicates that the synthesized GNPs are densely incorporated in the film. The size of GNPs
seems to be regulated. Such size regulation may be explained in terms of immediate captureof reduced gold by growing film. The film formation process can be controlled by means of
the concentration of HAuCl4 and gelatin and discharge duration, which affects the size andconcentration of GNPs in the membrane.
References:[1] T. Shirafuji, et al., Jpn. J. Appl. Phys. 2013, 52, 126202.
[2] H. Furusho, et al., J. Photopolym. Sci. Technol. 2007, 20, 229.

37

P-2

Iridium-Catalyzed Aerobic Dehydrogenative/Decarbonylative
Coupling of Salicylaldehydes with Alkynes
Shintaro Yamane,1 Tomoaki Hinoue,2 Yoshinosuke Usuki,1 Masumi Itazaki,1
Hiroshi Nakazawa,1 Yoshihiro Hayashi,3 Susumu Kawauchi,3Masahiro Miura,2 Tetsuya Satoh1
1
Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku,
Osaka 558-8585, Japan.
2
Guraduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
3
Department of Chemical Science and Engineering, Tokyo Institute of Technology,
2-12-1 (E4-6), Ookayama, Meguro-ku, Tokyo 152-8552, Japan.
Abstract: Transition-metal-catalyzed dehydrogenative coupling reactions have been regarded
as useful synthetic tools to prepare useful organic molecules from readily available starting
materials. In particular, the coupling of aromatic substrates possessing a heteroatom-containing
directing group with internal alkynes has been utilized for constructing fused heteroarenes. We
previously demonstrated that salicylaldehydes undergo dehydrogenative coupling with internalalkynes in the presence of a rhodium catalyst and a copper salt oxidant to produce 2,3disubstituted chromone derivatives.[1] In this study, we succeeded in finding thatdehydrogenative, decarbonylative coupling of the same combination of substrates can be
carried out efficiently by using an iridium catalyst together with a copper cocatalyst in place ofthe rhodium system under O2 (1 atm) to give 2,3-disubstituted benzo[b]furan derivatives.
Benzo[b]furan skeletons can be seen in various synthetic intermediates for pharmaceuticals andbiologically active compounds and functional organic materials. The present procedureprovides a straightforward, simple method for constructing such important structures.

References[1] Shimizu, M.; Tsurugi, H.; Satoh, T.; Miura, M. Chem. Asian J. 2008, 3, 881.

38

P-3

Photoswiching of Birefringence of Diarylethene Single Crystals
by Photochromic Reaction
Kohei Morimoto, Hajime Tsujioka, Daichi Kitagawa, Seiya Kobatake
Department of Applied Chemistry, Graduate School of Engineering,Osaka City University, Osaka 558-8585, Japan
Abstract: Photochromic compounds have drawn muchattention because the chemical and physical propertiesare changed by photoirradiation. Diarylethenes are oneof the most promising photochromic compounds
because of their high fatigue-resistance and high Figure 1. Diarylethenes used in this work.
thermal stability of two isomers[1]. In this work, we investigated the photoinduced birefringencechange of 1a and 2a single crystals. The transmittance spectra of the crystals under crossed
Nicols exhibited interference due to birefringence. Fig. 2 shows the interference color change
under crossed Nicols and the real color change under open Nicol in 2a crystal by ultraviolet(UV) light irradiation. The transmittance spectrum due to the interference color was shifted tolonger and shorter wavelength in 1a and 2a, respectively, as shown in Fig. 3. The results
indicate that the birefringence waschangedbythephotoisomerization reaction. Thechange in birefringence values of
the crystal was obtained by fitting
the theoretical formula to thetransmittance spectrum. The
single crystals of 1a and 2a
exhibitedthebirefringenceincrease and decrease upon
irradiation with UV light,
respectively. The relationshipbetweenthephotoinduced
birefringencechangeofdiarylethene crystals and theirmolecular
structureswasdiscussed based on the molecularpolarizability anisotropy.

Figure 2. Interference color change under crossed Nicols and realcolor change under open Nicol of 2a crystal (thickness: 5.1 μm).

Figure 3. Transmittance spectra of (a) 1a crystal (thickness: 45.2μm) and (b) 2a crystal (thickness: 5.8 μm) under crossed Nicols,
before UV irradiation (solid line) and after UV irradiation (dashedline).

References:[1] M. Irie, T. Fukaminato, K. Matsuda, S. Kobatake, Chem. Rev. 2014, 114, 12174−12277.

39

P-4

Development of Covariance Mapping Method for theInvestigation of Coulomb Explosion Processes
Tomomasa Nishimura, Masako Itsukashi, Tomoyuki Yatsuhashi
Graduate School of Science, Osaka City University, 3-3-138 Sugimoto,Sumiyoshi-ku, Osaka 558-8585, Japan
Coulomb explosion is one of the most extreme fragmentation process in chemistry due to thedestructive nature of multiply charged molecular cations [1]. We have investigated therelationship between the molecular structure and the ejection of fragment ions by Coulomb
explosion for various molecules by using a time-of-flight (TOF) spectroscopy [2]. The
qualitative interpretations of ion emissions even for large molecules can be made to someextent; however, the correlations between fragment ions are not known. In this study, wedeveloped the covariance mapping technique, which can be utilized to determine correlations
between fragment ions from TOF data. We prepared a data collection and analysis software,
which can process 8×1011 data within 1 hour. The experimental conditions were optimized forthe investigation of Coulomb exploison of diatomic molecules.
Multiple ionization of N2 was performed with a femtosecond laser pulse (0.8 µm, 40fs) and the ions formed by Coulomb explosion were detected by a linear-type TOF mass
spectrometer. The pressure of N2 was mainteined in the 10−6 Pa range (ca. 4×10−13 mol dm−3)
to avoid false covariance events. Figure 1 shows a covariance map showing the Coulomb
explosion of N2. The bottom and left traces
represent average TOF spectra. The typical featureof an ion generated by Coulomb explosion is thepresence of split peaks that are a consequence ofthe fact that ions with certain kinetic energies are
emitted along both the forward (f) and backward(b) direction relative to the ion flight axis. The
strong diagonal line in the map corresponds to theauto-correlations. On the other hand, four peaks
indicated by dashed circles show the correlation of
Nf3+ and Nb3+ , Nf2+ and Nb3+ , Nf3+ and Nb2+ ,and Nf2+ and Nb2+ , respectively. We conclude thatthe origin of the respective fragment ion pairs isdefinitively identified as being N26+, N25+, N25+,
and N24+.

Figure 1 A covariance map showing the ion
correlations in the Coulomb explosion of N2.

References[1] Yatsuhashi, T.; Nakashima, N. J. Photochem. Photobiol. C 2018 in press.
[2] Tanaka H.; Nakashima, N.; Yatsuhashi, T. J. Phys. Chem. A 2016, 120, 6917.

40

P-5

Synthesis of Single-Nano-Sized Iron Nanoparticles
from Liquid/Liquid Dispersion System
by Femtosecond Laser Irradiation
Yuki Horikawa, Takuya Okamoto, Tomoyuki Yatsuhashi
Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka
558-8585, Japan
Iron nanoparticles (FeNPs) have come to occupy an important position in magnetic andbiomedical materials. The production of FeNPs have been reported by using nanosecond
ultraviolet lasers, whereas femtosecond laser was rarely used [1]. In this study, we report thesynthesis of FeNPs by using water and n-hexane mixed solvent to control the size of FeNPs aswell as to remove the unwanted carbon aggregates.
We chose ferrocene as a reactant, andn-hexane or water and n-hexane mixture as
solvent. The reactant solution was exposed
to focused femtosecond laser pulses (0.8 µm,
40 fs, 0.4 mJ, 1 kHz) for 10 min.
Transmission electron microscopy (TEM)
was used to investigate the morphology andsize of FeNPs. When n-hexane was used asa solvent, FeNPs as well as carbon
aggregates were formed in abundance. The
mean and maximum diameters of FeNPs
were 10 nm and 50 nm, respectively(Fig.1a). On the other hand, we obtained
dispersed FeNPs with negligible amout of

Fig.1 TEM images and paticles size
distribution of FeNPs collected from (a)n-hexane solution and (b) water layer of waterand n-hexane mixture.

carbon aggregates when water and n-hexane mixture was used as solvent (Fig.1b). It isemphasized that single-nano-sized FeNPs are dominantly formed. The mean and maximum
diameters of FeNPs were 4.6 nm and 10 nm, respectively. We conclude that heterogeneous
reaction environment (n-hexane droplets in water) regulates the growth of FeNPs. In addition,hydrophobic carbon aggregates originating form cyclopentadienyl ligands were separated intoa hydrocarbon phase [2].
References[1] Wesolowski, M. J.; Kuzmin, S.; Wales, B.; Sanderson, J. H.; Duley, W. W. J. Mater. Sci.
2013, 48, 6212.
[2] Okamoto, T.; Mitamura, K.; Hamaguchi, T.; Matsukawa, K.; Yatsuhashi, T.
ChemPhysChem 2017, 18, 1007.

41

P-6

Stereoselective Synthesis of the Tripeptide of SF-2132
Akira Sawai, Ai Sekihara, Yoko Yasuno, Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
SF-2132 (1), isolated from the culture broth of Nocardiopsis sp. SF-2132, exhibits
antimicrobial activities against mycobacteria, Gram-positive bacteria, and Gram-negative
bacteria.[1] This hexapeptide contains four unusual amino acids. Among them, the structuresof the β-arginine and the dehydroamino acid moieties have not been established yet. In thispresentation, we would like to report the stereoselective synthesis of the tripeptide unit E-2
and Z-3. E-tripeptide 2 was synthesized via the Negishi cross coupling reaction of iodo
dehydroamino acid 4 under the mild condition.[2] Stereoselective synthesis of Z-isomer 3 was
furnished by the selective dehydration reaction of 5 via a cyclic sulfamidite.[3] Comparison of
NMR data of E-2, Z-3 and 1 suggested that the olefin geometry of 1 was putatively assigned
to be Z-configuration.

[1] Kondo, Y.; Tohyama, H.; Shomura, T.; Sezaki, M.; Niwa, T.; Kojima, M. Meiji Seika
Kenkyu Nenpo, 1985, 24, 27.
[2] Yasuno, Y.; Nishimura, A.; Yasukawa, Y.; Karita, Y.; Ohfune, Y.; Shinada, T. Chem.
Commun. 2016, 52, 1478.
[3] Stohlmeyer, M. M.; Tanaka, H.; Wandless, T. J. J. Am. Chem. Soc. 1999, 121, 6100.

42

P-7

Annealing temperature dependency of directly-bonded SiC/Si
junction characteristics
K. Shimozato, J. Liang, and N. Shigekawa
Graduate School of Engineering
Osaka City University
Sugimoto 3-3-138, Sumiyoshi, Osaka 558-8585, Japan
E-mail：m17tb029@ka.osaka-cu.ac.jp
M. Arai
New Japan Radio Co. Ltd.
temperature at the first (lowertemperature) step in the bonding process.
2.0x1015

1/C2 (cm4/F2)

We measured electrical characteristics
and die shear strength of directly-bonded
n—SiC(1×1017cm3)/p+-Si junction with theemphasis on their dependence on theannealing condition during bonding. As
reported for GaAs/GaAs junction[1], theelectrical characteristics of directlybonded junction are varied by annealing
conditions. In this work, we separated
annealing process into two steps. We
performed lower-temperature annealingat the first step and higher-temperature
one at the second step. Figure 1 shows
capacitance-voltage (C-V) characteristics
of junction prepared with different firststep annealing temperatures. The
impurity concentration obtained fromeach C-V curve is also shown in this figure.
Figure 2 shows the dependence of bonding
strength on the annealing temperature in
the first step. A concentration close to thatlayersobtainedbyforn--SiCcharacterizing Schottky junctions isobserved
forhigherannealing
temperatures in the first step. With
increasing annealing temperature at thefirst step, the impurity concentrations
tend to approach the concentration of SiC
and the bonding strength shows a highnumerical value. These results show
directly-bonded
SiC/Sijunctioncharacteristics depend on the annealing

1.5x1015

RT(26oC)
200oC
300oC

5.49x1016

1.0x1015

8.76x1016

5.0x1014
0.0

9.96x1016
-3

-2

-1

0

1

2

3

4

Voltage (V)Fig. 1 C-V characteristics of n-4H-SiC/p-Si junctions.
Bonding Strength (MPa)

Abstract:



16

15

14

13

0

50

100

150

200

250

300

350

o

Temperature ( C)

Fig. 2 Bonding
junctions.

strengthof

SiC/Si

References:[1]M. J. Jackson, et al. J. Appl. Phys. 110,
104903 (2011).

43

P-8

Light-induced hydrogen production by photosystem I-Pt nanoparticle
immobilized in porous glass plate nanopores
Makoto Hirano,1 Tomoyasu Noji,2 Keisuke Kawakami,2 Teturo Jin,3 Masaharu
Kondo,4 Hirozo Oh-oka,5 Nobuo Kamiya,2,1
1
Graduate School of Science, Osaka City University, 2 The OCU Advanced
Research Institute for Natural Science & Technology (OCARINA), Osaka City University,
3
Inorganic Functional Materials Research Institute, National Institute of Advanced Industrial,
Science and Technology (AIST),4Department of Life Science and Applied Chemistry,
Graduate School of Engineering, Nagoya Institute of Technology, 5Department of Biological
Sciences, Graduate School of Science, Osaka University,

Abstract: Artificial photosynthesis that converts solar energy into chemical energy, has
attracted a lot of attention as one of answers to solve the energy problem. Natural
photosynthesis is efficiently achieved by pigment-protein complexes, named photosystem I(PSI) and photosystem II. PSI produces the high reducing power to convert carbon dioxide tosugar using solar energy. Utilization of natural photosynthetic proteins for the construction of
artificial devices is an attractive approach, because the high quantum efficiencies oflight-induced charge separation are near 100%. For example, a light-induced hydrogenproduction system has been reported using a conjugate of PSI and Pt nanoparticle (PSI-PtNP)[1]. More recently, the hydrogen production system working even under aerobic condition has
been constructed by immobilizing the PSI-PtNP and cytochrome c6 (cyt c6) in a nanoporous
glass plate (PSI-PtNP/PGP) with a pore diameter of 50 nm [2]. The hydrogen evolution
activity of the PSI-PtNP/PGP was low because the reduction rate of PSI by cyt c6 was limitedby the cyt c6 diffusions in the PGP. In this study, we optimized the method immobilizing the
PSI-PtNP and cyt c6 into PGP. As a result, the maximum rate of the hydrogen production was
faster 6.5-times than that of the previous report. Furthermore, we investigated photochemical
activity of PSI inside PGP by measuring fluorescence spectra and flash-induced transientabsorption changes. We will discuss the relation between the electron transfer rates in PGP
and the hydrogen evolving efficiencies.
References:[1] L. M. Utschig, N. M. Dimitrijevic, O. G. Poluektov, S. D. Chemerisov, K. L. Mulfort andD. M. Tiede, J. Phys. Chem. Lett., 2011, 2, 236-241.
[2] T. Noji, T. Suzuki, M. Kondo, T. Jin, K. Kawakami, T. Mizuno, H. Oh-oka, M. Ikeuchi, M.
Nango, Y. Amao, N. kamiya, T. Dewa, Res. chem. Intermed. 2016, 42, 7731-7742

44

P-9

Dehydrogenative Coupling of Aromatic Carboxylic Acids andUnsaturated Compounds under Rhodium Catalysis○Asumi Sakai,1 Takeshi Okada,2 Tetsuya Satoh,1 Yoshihiro Hayashi,3
Susumu Kawauchi,3 Masahiro Miura2
1
Department of Chemistry, Graduate School of Science, Osaka City University, 3-3-138
Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan
2
Graduate School of Engineering, Osaka University, 2-1
Yamagaoka, Suita 565-0871, Japan3
Department of Chemical Science and Engineering, Tokyo Institute of Technology, 1-12-1
Ookayama, Meguro-ku, Tokyo 152-8552, JapanThe transition-metal catalyzed dehydrogenative coupling reactions of aromatic substrates
possessing a hetero atom-containing directing group with unsaturated compounds via
regioselective C-H bond cleavage are powerful synthetic tools for constructing π-conjugatedmolecules. Among directing groups, a carboxylic group coordinates to a metal center weekly
to allow smooth dissociation and coordination. Previously, our group has reported that thedehydrogenative coupling of benzoic acids with unsaturated compounds such as alkenes andalkynes can be conducted effectivity in the presence of a rhodium catalyst and a suitable
oxidant through ortho C-H bond cleavage [1].
In this work, we have found that the dehydrogenative coupling of indole-3-carboxylic acids
with alkenes proceeds involving C-H activation at their 4-position . Futhermore, thecarboxylic function of the 4-alkenylated products can be removed upon treatment with a CuO
catalyst [2] to produce 2,3-unsubstituted 4-alkenylindole derivatives.

[1] K. Ueura, T. Satoh, M. Miura, Org. Lett. 2007, 9, 1407-1409.
[2] L. J. Goossen, G. Deng, L. M. Levy, Science. 2006, 313, 662-664.

45

P-10

Synthesis, Properties, and Reactivities of(Nitronyl Nitroxide-2-ide)Copper Complex
Bearing Phenanthroline ligand
Kiyomi YAMADA1, Shuichi SUZUKI2, Masatoshi KOZAKI1,3, Keiji
OKADA1,3
1
Graduate School of Science, Osaka City University, 2 Graduate School of Engineering
Science, Osaka University, 3 Osaka City University Advanced Research Institute for Natural
Science and Technology (OCARINA), Sugimoto, Osaka 558-8585.
Radical metalloids have a metal-carbon bond at the C2-position in
NN (nitronyl nitroxide), and only a few studies have been reported
mainly from our laboratory. Recently, we reported that a Pt complex
exhibited a large shift in the oxidation potential of the NN moiety to thenegative direction by ~0.6 V compared to that of the parent NN(NN-H) [1]. Furthermore, an Au complex was successfully used as astable reagent in the Pd(0)-mediated cross-coupling reaction with aryl
iodides (ArI) to afford aryl-substituted NNs (Ar-NNs) in excellent
yields [2]. In order to develop further variation of these properties, wehave developed various radical metalloid reagents. In this study, we
designed and synthesized the Cu(I)-NN complex bearing 1,10-phenanthroline (1).
The copper complex 1 could be readily prepared in high yield and turned out to be stable insolid under aerated condition. The structure of 1 was confirmed by MS, ESR, and X-ray
crystal structure analysis. The copper complex 1 has a extremely low oxidation potential of
the NN moiety (0.56 V vs Fc/Fc+), which is ~ 1 V negative compared to that of NN-H. The
cross-coupling reaction of 1 with aryl halides proceeded in good yields in the presence ofPd2(dba)3•CHCl3
and
DPPF
Scheme 1.
(1,1'-bis(diphenylphosphino)ferrocene) (Scheme1). Aryl iodides were more reactive than aryl
bromides for both of electron-rich and -poor aryl
halides. Moreover, the reactions proceeded withheteroaromatic systems in good yields.
References:[1] Zhang, X.; Suzuki, S.; Kozaki, M.; Okada, K. J. Am. Chem. Soc. 2012, 134,
1786617868.
[2] Tanimoto, R.; Suzuki, S.; Kozaki, M.; Okada, K. Chem. Lett. 2014, 43, 678680.

46

P-11

Stereoselective Syntheses of β-Hydroxy Amino Acids
by Isonitrile Aldol Reaction
Kazuma Fuku, Yoko Yasuno, Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
β-Hydroxy amino acids, such as β-hydroxyvaline and β-hydroxyisoleucine, are often
found as components of bioactive natural products. In this presentation, we would like toreport stereoselective syntheses of β-hydroxy-α-amino acids by the aldol reaction of isonitrile
1 possessing a camphorsulfonamide chiral auxiliary.
Isonitrile aldol reaction of 1 with ketone 2 in the presence of 5 mol% of copper catalyst 3
and Et3N in dichloroethane (DCE) at 0 oC provided 4 which were hydrolyzed to giveN-formyl-β-hydroxy amino acid esters 5 [5872% yield (2 steps), dr = 2:1>10:1].[1] The
scope and limitation will be discussed.

[1] Shinada, T.; Oe, K.; Ohfune, Y. Tetrahedron Lett. 2012, 53, 3250.

47

P-12

One-pot Synthesis of Anthraquinones by Sc(OTf)3-catalyzed
Tandem Reaction
Yohei Toriyama, Yoko Yasuno, Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
A tandem reaction using a single catalyst has attracted much attention as an efficientsynthetic method.[13] In the last annual meeting, we reported one-pot conversion of 1 tocyclic compounds 4 in the presence of catalytic amount of Sc(OTf)3.[4] In this reaction
process, two different reactions, (i) dehydration and (ii) DielsAlder (DA) reaction were
promoted by Sc(OTf)3 in an efficient manner. During the course of our continuous studies, wefound that the DA product 4 underwent (iii) aromatization reaction in the presence ofSc(OTf)3. These results led us to investigate a new Sc(OTf)3-catalyzed tandem reaction
including three different reactions (i)(iii) in one-pot. Indeed, various anthraquinones listed
below were prepared from 1 and 2. The scope of the new tandem reaction will be discussed.

[1] Wasilke, J. C.; Obrey, S. J.; Baker, R. T.; Bazan, G. C. Chem. Rev. 2005, 105, 1001.
[2] Shindoh, N.; Takemoto, Y.; Takasu, K. Chem. Eur. J. 2009, 15, 12168.
[3] Camp, J. E. Eur. J. Org. Chem. 2017, 425.
[4] The Chemical Society of Japan 97 th Spring Annual Meeting, 2017, 3C6-35.

48

P-13

Total Synthesis of SF-2132
Ai Sekihara, Akira Sawai, Yoko Yasuno, Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
SF-2132 (1), isolated from the culture broth of Nocardiopsis sp. SF-2132, is a uniqueclass of unusual amino acid-rich peptides.[1] In the precedented paper, we reported thestereoselective synthesis of the tripeptide chain 5 including the dehydroamino acid moiety of
1.[2] These synthetic and spectroscopic analysis suggested that the dehydroamino acid moietyof the natural product has Z-configuration. In this presentation, we would like to report thefirst total synthesis of 1 or 2 by coupling of fragments (R)/(S)-3, 4 and 5. Stereochemistry ofthe β-arginine moiety of 1 and 2 will be discussed.

[1] Kondo, Y.; Tohyama, H.; Shomura, T.; Sezaki, M.; Niwa, T.; Kojima, M. Meiji Seika
Kenkyu Nenpo 1985, 24, 27.
[2] The Chemical Society of Japan 98 th Spring Annual Meeting, 2018, 1D2-17.

49

P-14

Long-lived Molecular Tetracations of
Fluorine-containing Aromatic Compounds
Akihiro Kitashoji, Tomoyuki Yatsuhashi
Department of Chemistry, Graduate School of Science, Osaka City University
Abstract: Multiply charged molecular cations (MMCs) have the potential to be a novel
chemical species due to their high electron affinity and charge-dependent reactions. However,
little is known about MMCs because they are quite unstable due to Coulombic repulsion. In
this study, we showed that intact tetracations of hexafluorobenzene (C6F6) andoctafluoronaphthalene (C10F8) were produced by femtosecond laser pulses. C6F64+ is thesmallest tetracation ever observed for aromatic compounds. We investigated the reactivity of
C6F6z+ and C10F8z+ (z = 1–4) by using the time-of-flight mass spectrometer equipped with an
ion gate and a curved-field reflectron.
Figure 1 shows the time-of-flight mass spectra of a) C6F6 and b) C10F8 ionized by afemtosecond laser pulse (0.8 m, 35 fs, >1014 W cm−2). Intact tetracations of C6F6 and C10F8
were clearly identified (see insets) in abundance. Each charge states of C6F6z+ weresuccessfully selected by a homebuilt Bradbury-Nielsen ion gate (gate width: 36.5±0.5 ns, riseand fall time: >3 ns) [1] as shown in Figure 2. Supposing that the fragmentation of MMCs
occurs in the first field-free drift region, the product ions are recorded at times that are
different from their precursor ions. However, such product ions were not observed for C6F6z+(z = 1–4) and C10F8z+ (z = 2–4). Therefore, the lifetimes were estimated to be more than 9 sfor C6F64+ and 11 s for C10F84+. We conclude that tetracations are potential candidates asnovel reactive species based on the relatively long lifetime and the expected high potential
energy (>50 eV).

Figure 1. Time-of-flight mass spectra ofa) C6F6 and b) C10F8.

Figure 2. Time-of-flight spectra of a single
charge state of C6F6z+ (z = 1–4) selected by
Bradbury-Nielsen ion gate. * indicates H2O+.

References:[1] Kitashoji, A.; Yoshikawa, T.; Fujihara, A.; Kamamori, T.; Nashima, S.; Yatsuhashi, T.
ChemPhysChem 2017, 18, 2007.

50

P-15

Synthetic Study of Phomopsin A
Yuma Karita, Yoko Yasuno, and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Phomopsin A (1) isolated from
Diaporthe toxica is known to be a
mycotoxin.[1] According to the complex
structural feature as well as its potent
microtubule depolymerization activity, 1 has
attracted much attention as a synthetic target.
Structure of 1 is characterized by theconstrained 13-membered cyclophane ring consisted of β-OH-DOPA, β-OH-Ile, and β,γ-ΔVal.
In this presentation, we would like to report our efforts toward the first total synthesis of 1 via
the cyclophane intermediate 2.
Three fragments 3[2], 4[3] and 5[4] were sequentially condensed to give cyclizationprecursor 6 which was subjected to macrocyclization reaction under the high dilution
conditions. After the optimization, cyclophane 2 was obtained in 83% from 6. Recent progresson the conversion of 2 to the target natural product will be reported.

[1] Culvenor, C. C. J.; Beck, A. B.; Clarke, M.; Cockrum, P. A.; Edgar, J. A.; Frahn, J. L.;
Jago, M. V.; Lanigan, G. W.; Payne, A. L.; Peterson, J. E.; Petterson, D. S.; Smith, L. W.;
White, R. R. Aust. J. Biol. Sci. 1977, 30, 269.
[2] The Chemical Society of Japan 97 th Spring Annual Meeting, 2017, 2C7-47.
[3] Forbec, E. M.; Evans, C. D.; Gilleran, J. A.; Li, P.; Joullié M. M. J. Am. Chem. Soc. 2007,
129, 14463.
[4] The Chemical Society of Japan 97 th Spring Annual Meeting, 2017, 2C7-36.

51

P-16
Template for the abstract

Hydrolysis of Organophosphates
Catalyzed by Plussian Blue Analogs    
Chihiro Terashima, Hiroyasu Tabe, Yusuke Yamada
Graduate School of Engineering, Osaka City University
3-3-138 Sugimoto Sumiyoshi-ku, Osaka-shi, 558-8585, JAPAN
Abstract: Prussian blue analogs (MN[MC(CN)6], MN = FeII/III, MnII, CoII, GaIII, MC = FeII,CoIII, RuII, IrIII, PtIV) were examined as heterogeneous catalysts for hydrolysis oforganophosphates. The catalytic activity of the Prussian blue analogs depends on not only MN
ions regarded as potential active sites but also MC ions with coordinatively saturated structure.
【Introduction】
Removal of organophosphates used as agricultural chemicals has been strongly demanded,because organophosphates are listed as the 2nd cause of poisoning accidents in Japan.
Organophosphates are currently removed by adsorbents such asactivatedcarbon,however,completeremovaloforganophosphates at high concentration can be hardly achieved.
Thus, catalytic decomposition of organophosphates is anotherpromising method for removal of organophosphate. We reportherein that Prussian blue analogs (PBAs, Fig. 1) exhibits catalytic
activity for the hydrolysis of organophosphorus compounds.
Fig. 1 Partial structure of【Experiments】
PBA
Heterogeneous catalysis of each PBA was evaluated in a buffer solution containing
4-nitrophenylphosphate disodium salt and stirred at 60 ° C.
【Results】
Catalytic activity of various PBAs (MN[MC(CN)6], MN =FeII/III, MC = FeII, PtIV, IrIII, CoIII, RuII) for hydrolysis of
4-nitrophenylphosphate (NPP) was compared based on theformation rates of 4-nitrophenol (NP) (Fig. 2). The initial
reaction rates of 86, 6.9 , 5.6, 2.6 and 1.7 × 107 M s-1 for
PBAs with MC = FeII, PtIV, IrIII, CoIII and RuII, respectively,
depend on the MC ions. The highest activity of Prussian
blue (MC = FeIII) resulted from thehigher oxidation sate of Fe ions at MN
sites in the mixed valence state.

52

Fig. 2 Time profiles of NP production by hydrolysis of
NPP (25 mM) catalyzed by K0.1[FeIII(H2O)1.4]1.3[FeII(CN)6] (●), FeIIx[MC(CN)6] (MC = CoIII (●), RuII (■),
IrIII (◆), PtIV (▲)) ([NPP]/[FeII/III] = 100) or no catalyst(■)
in
2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonate buffer (0.75 mL, 100 mM, pH 8.2) at 60 °C.

P-17

Synthetic Study of Deoxyustiloxin D
Kenta Sakai, Yoko Yasuno, and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Ustiloxin D (1) is a unique class of unusual peptides which is characterized by thepresence of a 13-membered cyclic core with an ether linkage.[1] This natural product reveals
potent inhibitory activities against tubulin polymerization. Based on the X-ray analysis of
ustiloxin D and tubulin complex,[2] we proposed that potent biological activity would bemaintained in the designed analog 2. In this presentation, we would like to report our
synthetic efforts.
Fragments of 3, 4, and 5 were sequentially combined. The resulting acyclic molecule
was subjected to macrocyclization reaction to provide 6. Conversion of 6 via coupling
reaction with 7 to provide the target analog 2 will be reported.

[1] Koiso, Y.; Natori, M.; Iwasaki, S.; Sato, S.; Sonoda, R.; Fujita, Y.; Yaegashi, H.; Sato, Z.
Tetrahedron Lett. 1992, 33, 4157.
[2] Ranaivoson, F. M.; Gigant, B.; Berritt, S.; Joullié, M.; Knossow, M. Acta Crystallogr. D
2012, 68, 927.

53

P-18

Improvement of water permeability ofa bone-regeneration scaffold by irradiating
atmospheric pressure plasma jet
1

Yuki Hamamoto, 1Masato Oshiro, 2Kumi Orita,
2
Yoshihiro Hirakawa, 2Hiromitsu Toyoda, and 1Tatsuru Shirafuji
1
Department of Physical Electronics and Informatics, Graduate School of Engineering,Osaka City University, Osaka, 558-8585 Japan2
Department of Orthopedic Surgery, Graduate School of Medicine,
Osaka City University, Osaka, 545-8585 Japan
1. Introduction
(a)
(b-1)
(b-2)
(b-3)
+ 0 ns+ 20 ns+ 40 ns
An atmospheric pressure plasma jet (APPJ)
employing dielectric barrier discharge ofhelium gas is known to be a source of“plasma bullets”, which propagate in(b-4)(b-5) + 100 ns
(b-6)
(b-7)
+ 60 ns+ 80 ns+ 120 ns
high-purity helium gas channels [1]. A
plasma bullet propagates to the direction
independent of gas-flow direction, andnicely separated when they encounterbranches of gas channels. These unique Fig. 1 Propagation of plasma bullets out of a glassfeatures of “plasma bullets” may be used filter.
for the treatment of internal surfaces of anjust after dropped
after 20 min
interconnected porous scaffold used in bone regeneration.
glass
filterbullets
exit

bac k
side

He APPJ

bulletsenter

fr ontside

bac k sideof glass
filter

2. Experimental procedure
We have irradiated a helium APPJ to a glass filter. The glass
filter is used as a substitute of a scaffold made of expensive
HA or β-TPC. The thickness and pore channel diameter of theglass filter are 3.15 mm and 160-250 μm. The scaffold washydrophobic treated using fluorinated compounds before
APPJ irradiation. Plasma bullets were injected from one sideof the scaffold. We have expected that the bullets penetrate thescaffold and ejected from the other side of it.

5 mm

back
side

front side(a) Hydrophobic glass filter
just after dropped

after 0.4 sec

back
side

front side(b) APPJ-treated glass filter

Fig. 2 Water permeability of theglass filter before and after theplasma-bullet treatment.

3. Results and discussion
Figures 1(b-1) to 1(b-7) show time-evolution of plasmabullets on the back side of the APPJ-irradiated glass filter, which indicates that the bullets
penetrate the glass filter and exit from its back side. Figures 2(a) and 2(b) show that water
permeability of the back side of the glass filter before and after the APPJ treatment. These
results indicate that the internal surfaces of channels in the glass filter have becomehydrophilic by propagation of plasma bullets through the scaffold.
References:[1] M. Teschke, J. Kedzierski, E. G. Finantu-Dinu, D. Korzec, and J. Engemann, IEEE Trans.
Plasma Sci. 2005, 33, 310.

54

P-19

Characterization of damages in n-Si
due to irradiation of Ar fast atom beams
Y. Matsumoto, K. Hisamoto, J. Liang and N. Shigekawa
Graduate School of Engineering
Osaka city University,Japan
Sugimoto 3-3-138,Sumiyoshi,Osaka 558-8585, Japan
a14tmX0E36@st.osaka-cu.ac.jp
Abstract:
Surface-activated bonding (SAB) technologies have widely been applied for bonding
semiconductors[1]. The SAB process includes irradiation of Ar fast atom beams (FAB), whichplays a role of activating surface of semiconductor. It is noted that damages are formed at thebonding interfaces by the FAB irradiation. The damages can be healed by annealing interfaces
at low temperatures[2]. However, the temperature and duration of annealing are not yet
optimized since they are likely to depend on the depth and density of damages[3].
We investigated the depth of damages due to irradiation of Ar FAB. Irradiated n-Si was
etched by TMAH and Au/n-Si Schottky diodes were fabricated on unveiled Si surfaces. The
acceleration voltage and irradiation time were 1.4 kV and 600 s, respectively. We measured
current-voltage (I-V) and capacitance-voltage (C-V) characteristics of Schottky diodes.
Figure 1 shows the dependence of Schottky
barrier height (SBH) that were derivatedfrom the I-V characteristics on the etching
depth. We found that SBH was decreased
from 0.75 to 0.63 eV by the FAB irradiation.
SBH was recovered by etching the irradiatedn-Si surfaces to a depth of ≈ 60 nm. We
observed a similar relation in SBH obtained
from the C-V measurement. Their results
suggested that damages were formed in a ≈60–nm thick region of the n-Si surface andthey lowered SBH of Au/n-Si junctions.

References:[1] J. Liang, et al. Jpn. J. Appl. Phys. 54, 030211 (2015).
[2] S. Hisamoto, et al. IMFEDK, pp. 40-41 (2016).
[3] T. Seki, et al. Nucl. Instrum. Methods Phys. Res. B 164-165, 650 (2000).

55

P-20

Synthesis and Properties of SolubleBis(trioxytriphenylamine) Derivatives
Kento Shimode,1 Shuichi Suzuki,2 Masatoshi Kozaki,1,3 Keiji Okada1,3
1
Graduate School of Science, Osaka City University, 2Graduate School of Engineering
Science, Osaka University, 3Osaka City University Advanced Research Institute for Natural
Science and Technology (OCARINA), Sugimoto, Osaka 558-8585.
Abstract:
Generally, radical cation species of triphenylamine derivatives without p-substitutents are
unstable and undergo dimerization because of the large spin density at the para-positions. We
have designed and synthesized oxygen-bridged triphenylamine (trioxytriphenylamine: TOT).
We sucessfully prepared its radical cation and radical-substituted TOT radical cations andclarified their unique magnetic properties. [1] More recently, we synthesized a TOT-dimer(bisTOT), and investigated magnetic properties of their dicationic species [2]. However, theradical cation species of bisTOT was difficult to obtain because of 1) the proximate first andsecond oxidation potentials (EOX1 = +0.10 V, EOX2 = +0.20 V, vs Fc/Fc+, EOX = 0.10 V) and2) low solubility of the neutral and dicationic species. In order to obtain insight into
TOT-dimer radical cation species, we designed and synthesized TOT-dimers 1 and 2.
Compound 1 and 2 were prepared via long steps (6 and 7 steps with an overall yield of 12%
and 9 %, respectively) from known 1-bromo-3-methoxy-5-pentylbenzene as a starting
compound. Compound 1 and 2 exhibited good solubility in usual organic solvents such as
CH2Cl2 and toluene. Compound 1 showed a larger separation of oxidation potentials (EOX1 =–0.03 V, EOX2 = +0.13 V vs Fc/Fc+,EOX = 0.16 V) in benzonitrile compared with bisTOT.
From the absorption spectra and ESR spectra in CH2Cl2, radical cation species of 1 and 2
were suggested to be in a mixed-valence state of Class II and Class II/III border, respectively.
We could succesfully isolate radical cation 1•+ in a pure form.

[1] Okada, K. et al. Angew. Chem., Int. Ed. 2005, 44, 4056–4058. Wang, X. et al. J. Am. Chem.
Soc. 2013, 135, 1491214915. Okada, K. et al. Chem. –Asian. J. 2012, 7, 1604–1609. Okada,K. et al. Angew. Chem., Int. Ed. 2012, 51, 3193–3197.
[2] Okada, K. et al. Eur. –J. Chem. 2017, 23, 16014–16025.

56

P-21

Identification of S. pombe genes involved in spore maturation

Bowen ZHANG1, Yuhei TAHARA1,2, Makoto MIYATA1,2
and Taro NAKAMURA1,2
1) Graduate School of Science, Osaka City University, Japan2) The OCU Advanced Research Institute for Natural Science and Technology (OCARINA),
Osaka City University, JapanThe fission yeast Schizosaccharomyces pombe form spores in response to nitrogenstarvation. The spores are dormant cells with high resistance to various stresses, such asheat, digestive enzymes and organic solvents. Spore maturation is a result of significant
cellular structure morphological changes, mainly including spore wall formation andsimultaneous outer forespore membrane (FSM) autolysis. The spore wall is critical forresistance of spores to the external stresses and it is formed in the lumen of the FSM, adouble layer compartment surrounding a spore. During spore maturation, the inner layer of
the FSM becomes the future plasma membrane and the outer one disappears ultimately. By
random mutagenesis, the spoE22 strain was originally isolated as a mutant showing defectin the outer layer FSM autolysis. Under an electron microscope, the surface of the spoE22
spores was also found to be abnormally smooth, without spike structure seen in wild typespores. This study demonstrated that this defect of outer FSM autolysis was caused by theabsence of Meu5, an RNA binding protein whose expression is upregulated during meiosis[1, 2]. Meu5 binds to poly(A) domain, stabilizing the mRNAs of more than 80 target genes.
To screen the genes that are directly involved in the outer FSM lysis, we observed the FSMof the strains in which the Meu5 target gene is disrupted. According to the observation result,several genes were supposed to be related to outer FSM autolysis.
1. Amorim M.J.; Cotobal C; Duncan C; Mata J. Global coordination of transcriptional
control and mRNA decay during cellular differentiation. Molecular Systems Biology
2010, 6:380
2. Watanabe T.; Miyashita K.; Saito T.T.; Yoneki T.; Kakihara Y.; Nabeshima K.; Kishi
Y.A.; Shimoda C.; Nojima H. Comprehensive isolation of meiosis-specific genes
identifies novel proteins and unusual non-coding transcripts in Schizosaccharomyces
pombe. Nucleic Acids Research 2001, 29(11): 2327-2337.

57

P-22

The Short Step Synthesis and Properties of Triazapyrene
Derivatives
Yuta Omura,1 Yoshimitsu Tachi,1 Keiji Okada,1,2 Masatoshi, Kozaki1,2
1Graduate School of Science, Osaka City University, 2Osaka City University,
Advanced Research Institute for Natural Science and Technology (OCARINA)
Abstract: Introducing multi-nitrogen atoms into polycyclic aromatic hydrocarbons is effective
methods for developing novel n type organic semiconductors. However, there are limited
number of reports of nitrogen-rich polycyclic aromatic compounds due to the lack of aneffective synthetic method. In our research, using pyrene as a framework, we developed a shortstep synthetic method for triazapyrene derivatives that involves Pd-catalyzed cross-coupling
reactions via C–H activation and Cu-catalyzed annulation reactions.[1]Scheme 1. Synthesis of triazapyrene 1

The direct palladium-catalyzed C–H substitution was carried out using 1,3-dicyanobenzene
and 4-bromopyridine hydrochloric acid salt to afford 4-(2,6-dicyanophenyl)pyridine (2). The
reaction of 2 with Grignard reagents and the Cu-catalyzed C-N bond formation produced 1 in
44% yield (Scheme 1). Both absorption and emission bands of triazapyrene 1 with threepyridine rings observed in longer wavelength region compared to the corresponding bands ofpyrene. While reduction wave of pyrene
was not observed, triazapyrene 1 shows
reversible reduction waves at 1.94 V vsFc/Fc+. Crystal structural analysis
establishes that the large dihedral angles
between the pyrene core and the p-tolyl
group (51o and 59o) and the planar
pyrene cores have π-stack packing. The
detailed

synthesis

andproperties

triazapyrene derivatives will be reported.

ofFigure 1. Packing structure of triazapyrene 1[1] Ihanainen, N. E.; Kumpulainen, E. T. T.; Koskinen, A. M. P. Eur. J. Org. Chem. 2015,
32263229. Zhang, L.; Ang. G. Y.; Chiba, S. Org. Lett. 2010, 12, 36823685.

58

P-23

photo
    

Visible light-driven carbon-carbon bond formationfrom CO2 as a feedstock with the system
containing malic enzyme and dye
Takayuki Katagiri1, Shusaku Ikeyama2, Yutaka Amao1,2,3
1
Graduate School of Science, Osaka City University,

2

The Advanced Research Institute for Natural Science and Technology, Osaka City University
3

Research Center for Artificial Photosynthesis (ReCAP), Osaka City University

Recently, solar energy based CO2 utilization technologies including artificial photosynthesis
have been received much attention. As one of these technologies, CO2 conversion system
consisting of photosensitizer and catalyst were constructed. To use CO2 as a feedstock, effective
catalyst for carbon-carbon bond formation from CO2 is required. For example, malic enzyme(ME) catalyzes a reaction of introducing CO2 as a carboxy group to pyruvate (C3 compound)to form malate (C4 compound) via oxaloacetate in the presence of natural co-enzyme NADPH.
More recently, we reported that multi-electron reduced form of diphenylviologen (PV) act as
an artificial co-enzyme for ME [1,2].
In this study, novel water soluble viologen, 1,1'-bis(p-sulfonatophenyl)-4,4'-bipyridiniu
dichloride (PSV) based on PV, was synthesized as an artificial co-enzyme for ME. The
electrochemical and photoreduction properties of PSV were investigated. Quenching ofphotoexcited state of water soluble zinc porphyrin (ZnP) was also studied using fluorescence
spectroscopy. In addition, PSV was applied for visible light-driven oxaloacetate and malate
production system.
The first and second reduction potential of PSV were -0.38 and -0.72 (vs Ag/AgCl),
respectively. The photoreduction properties of PSV in the visible light-irradiation withphotosensitization of ZnP. As a result of photo-irradiation, the absorption maximum at 650 and
710 based on two-electron reduced form of PSV (PSV0) was increased with irradiation time.
When a sample solution containing triethanolamine (TEOA), water soluble porphyrin (P), PSV,
pyruvate, Mg2+ and ME in CO2 saturated pH 7.4 bis-tris buffer was irradiated with visible-light,
oxaloacetate and malate were produced. Thus, visible light-driven carbon-carbon bondformation system was constructed with ME and PSV.

2Cl-

Fig.1 Visible light-driven malate production frompyruvate and CO2 via oxaloacetate with ME.

2Na+

Fig.2 Chemical structure of 1,1’-bis(psulfonatophenyl)-4,4’-bipyridinium salt

[1] Y. Amao, S. Ikeyama, T. Katagiri, K. Fujita, Faraday Discuss., 2017, 198, 73[2] T. Katagiri, S. Ikeyama, Y. Amao, J. Photochem. Photobiol. A, 2018 in press

59

P-24

Allosteric effect of a zinc porphyrin receptor
with sterically bulky shielding units
Tomoaki, Nishimura;1 Yoshito, Sasaki; 1 Yoshimitsu, Tachi; 1
Keiji, Okada; 1,2 Masatoshi, Kozaki1,2
1Graduate School of Science, Osaka City University, 2Osaka City University, Advanced
Research Institute for Natural Science and Technology (OCARINA)
Abstract: Artificial allosteric systems are attractive as intelligent receptors and biomimetic
catalysts. Only a few studies have been reported allosteric systems using organic molecules asexternal sitmuli1. We prepared zinc porphyrin receptor 1 with sterically bulky shielding units(HPhB) to allosterically regulate ligand-binding ability using chemical stimuli. Receptor 1
recognizes a specific guest molecule through multiple supramolecular binding sites. The
formation of 1•2 would reduce the binding constant between diethynylporphyrin (DEP) and 4phenylpyridine (PhPy) because of the shielding of DEP by HPhB.
UV-vis spectral titration in toluene suggested that 1 and 2 formed stable 1:1 complex 1•2
through hydrogen bonds between a diamidopyridine (DAP) and uracil (U) unit andcoordination bonds between tetraphenylporphyrin (TPP) and pyridine units (Py). The binding
constant between DEP in 1•2 and PhPy was 0.089 times smaller than that between 1 and PhPy.
These results suggest that ligand-binding ability of DEP in 1 was allosterically inhibited by theformation of 1•2. We will present molecular design and the allosteric effect in detail.

References[1] Sasaki, Y.; Suzuki, S.; Okada, K.; Kozaki, M. Tetrahedron Lett. 2016, 57, 40824085.

60

P-25

Fabrication ofsemiconductor/patterned metal layer junctions
T. Hishida, S. Kohno, J. Liang, and N. Shigekawa
Graduate School of Engineering
Osaka City University
Sugimoto 3-3-138, Sumiyoshi, Osaka 558-8585, Japan
E-mail: a14tme0g29@ex.media.osaka-cu.ac.jp
Abstract:
III-V-on-Si multijunction solar cells are promising as high-efficiency and low-cost
photovoltaics. We previously fabricated InGaP/GaAs//Si triple-junction cells by using
surface-activated bonding (SAB) and achieve a high efficiency (~26%) [1]. However, it was
found that damages were introduced to the bonding interface by Ar beam irradiation in thesurface-activating process. This led to the increase in the interface resistance [2]. A practicalsolution is likely to be provided by using patterned ohmic metal with passivation layer on the
Si bottom cells.
In this work, we successfully fabricated a p+-Si // patterned Al with SiO2 / p+-Si junction by
SAB of a p+-Si substrate and a patterned Al layer and a subsequent annealing (500℃ 1hour).
The resistivity of p+-Si substrate was 0.002-0.005 Ω·cm. A schematic cross section and ascanning electron microscope (SEM) image of the junction are shown in Fig. 1 and 2,
respectively. Figure 3 shows the current-voltage (I-V) characteristics of the junction. Excellent
ohmic properties were confirmed.

Current density(A/cm2)

30
20

R = 0.004-0.01 Ω・cm2
10
0
-10
-20
-30
-0.10

-0.05

0.00

0.05

0.10

Voltage(V)Fig.1. A schematic crossFig. 2. An SEM image of

Fig. 3 I-V characteristics ofsection of a p+-Si/patterned

a p+-Si/patterned Al with

a p+-Si/patterned Al with

Al with SiO2/p+-Si junction.

SiO2/p+-Si junction.

SiO2/p+-Si junction.

References:[1] N. Shigekawa, et al. Jpn. J. Appl. Phys. 54, 08KE03 (2015).
[2] J. Liang, et al. Jpn. J. Appl. Phys. 54, 030211 (2015).

61

P-26

Analysis of neural connection between pineal organ and somebrain regions in zebrafish
Makomo Kakiya1, Seiji Wada1, Emi Kawano-Yamashita1, Mitsumasa
Koyanagi1, 2, Akihisa Terakita1, 2
1

Department of Biology and Geosciences, Graduate School of Science, Osaka City University,
Osaka, Japan; 2OCARINA, Osaka City University, Osaka, Japan
Many lower vertebrates capture light not only with eyes but also with pineal organs, which

are known as a melatonin-secretion organ in mammalian brains. Interestingly, pineal organsof teleosts can detect wavelength of lights in addition to light intensity. Several lines ofevidence suggested that pineal photoreceptive pigments, exorhodopsin and parapinopsin are
localized in different cells and are involved in the intensity detection and the wavelength
discrimination, respectively [1, 2]. Because previous studies suggest that neural responses ofthe photoreceptor cells transmit to the brain, analyses of neural projection of thephotoreceptor cells via ganglion cells is important to speculate physiological functions of thepineal photoreception.
Previous histological studies by anterograde-tracing suggested that light information
captured by the pineal photoreceptor cells might transmit into the several regions of brain
including mesencephalon via ganglion cells in zebrafish. However, it has not been uncovered
whether light information about the wavelength discrimination transmits to the regions. In
this study, we tried histological analysis to identify regions which the pineal photoreceptor
cells expressing parapinopsin project to, with some kinds of transgenic zebrafish.
[1] Kawano-Yamashita E.; Koyanagi M.; Terakita A. Springer 2014, 1-21
[2] Koyanagi M.; Kawano E.; Kinugawa Y.; Oishi T.; Shichida Y.; Tamotsu S.; Terakita A.
Proc. Natl. Acad. Sci. USA 2004 6687-91.

62

P-27

Photocatalytic activity of gallium oxynitride
for H2 evolution under visible light irradiation
Yuma Kato1, Muenaki Yamamoto2, Akiyo Ozawa1and Tomoko Yoshida3
13Graduate School of Engineering, Osaka City University
2
Graduate School of Engineering, Nagoya University

Advanced Research Institute for Natural Science and Technology, Osaka City University

Recently, we found that gallium oxide (Ga2O3) showed photocatalytic activity for water
splitting reaction under ultra violet light irradiation [1]. In order to generate visible light
response in Ga2O3 photocatalysts, we tried nitriding Ga2O3 in our previous study, however, H2evolution reaction did not proceed over the nitrided Ga2O3. In this study, gallium oxynitride(GaON) samples were prepared by nitriding gallium oxide hydroxide (GaOOH) and we
examined whether GaON samples promote H2 evolution from an aqueous methanol solution
under visible light irradiation.
GaOOH powder was obtained by autoclaving gallium nitrate aqueous solution at 120 ℃ for
6 h followed by filtration and drying. Nitriding of GaOOH was performed by calcination
under NH3 flow. Detailed nitriding conditions are summarized in Table 1.
Diffuse reflectance spectra (DRS) indicated that absorption bands appeared in the visible
light region after nitriding (Fig.1). N K-edge XANES and N 1s XPS spectra of the preparedsamples showed the peaks attributed to nitrogen species and the peaks became sharper withhigher nitriding temperatures. These results suggested that prepared samples actuallycontained nitrogen. In XRD measurement of the prepared samples, as nitriding temperature
exceeded 973 K, gallium nitride (GaN) phase started to appear, and the samples had higher
crystallinity with higher nitriding temperature. Similar results were obtained from the Ga
K-edge EXAFS measurement. H2 evolution reactions using prepared samples were performed
under visible light irradiation (λ>420 nm).This reaction did not proceed over GaN reference
sample while the sample (e) promoted H2 evolution.
249000

3.5

Table 1. Nitriding
conditionof GaOOH
Nitridingcondition

(a)
(b)
(c)
(d)
(e)
(f)
(g)2

time NH3 flow rate(h)(mL/min)
15
60
15
60
15
60
15
60
15
60
-

1GaOOH, 2GaN (reference samples)

199000

2.5

(c)

2

1.5

200

(g)

149000

(g)

(f)
(e)

99000

(d)
(c)

49000

(a)

0.5
0

(f)

(b)

1

(d)
(e)

Intensity (a.u.)

1

temp
(K)
773
873
973
1073
1173
-

Kubelka-Munk (a.u.)

Entry

3

(b)
300

400
Wavelength (nm)

500

600

Figure 1. DRS spectra of prepared samples(a-g) the samples described in Table 1.

-1000

(a)
25

30

35

40

45

2θ (degree)

50

55

60

Figure 2. XRD patterns of prepared samples(a-g) the samples described in Table 1.

[1] M. Akatsuka, T. Yoshida, N. Yamamoto, M. Yamamoto, S. Ogawa, S. Yagi, J. Phys. Conf.
Ser. 2016, 712, 012056.

63

P-28

Estimating spectral sensitivities of various opsins based on
biochemical responses of cultured cells expressing opsins
Tomohiro Sugihara1, Takashi Nagata1, Benjamin Mason2, Mitsumasa
Koyanagi1,3, Akihisa Terakita1,3
1Department of Biology and Geosciences, Graduate School of Science, Osaka City University,
Osaka, Japan; 2Department of Genetics, Stanford School of Medicine, Stanford University,
Stanford, CA, United States; 3OCARINA, Osaka City University, Osaka, Japan
Many animals capture light information by light sensitive proteins, opsins for various

photoreceptive functions.

Thousands of opsins have been identified from ocular andextraocular tissues of a wide variety of vertebrates and invertebrates.

Spectral sensitivity ofthe opsin provides basic and important information to speculate its relevant physiology but
spectral characteristics of not a few opsins still unknown. Basically, we express opsins in
cultured cells and obtain functional recombinant opsin-based pigments. However, lower
expression level of opsins often limits to analyze their spectral sensitivities by spectroscopic
method.
We recently focused on a light-induced second messenger change in cultured cells andhave succeeded in quantitatively analyzing wavelength-dependent cAMP changes in cultured
cells expressing opsins even in their low expression level. In this study, we showed that themethod based on cAMP changes is useful to obtain spectral sensitivities of opsins of which
absorption spectra have been unknown [1].
[1] Sugihara, T.; Nagata, T.; Mason, B.; Koyanagi, M.; Terakita, A. PLoS One 2016, 11(8)

64

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Contribution of parapinopsin “bistability” to light-responses
in the zebrafish pineal organs
Baoguo Shen1, Seiji Wada1, Emi Kawano-Yamashita 1, Mitsumasa
Koyanagi1, 2, Akihisa Terakita1, 2
1
Department of Biology and Geosciences, Graduate School of Science, Osaka City
University, Osaka, Japan; 2OCARINA, Osaka City University, Osaka, Japan
Pineal-related organs in lower vertebrates can discriminate UV and visible lights.
We previously found that a pineal-specific opsin, parapinopsin, which serves as a UV-sensitive pigment in the wavelength discrimination [1-3]. Parapinopsin has a uniquemolecular property called “bistable nature” different from that of visual opsin: upon
UV-light absorption, parapinopsin converts to a stable photoproduct, which reverts tothe dark state upon subsequent visible light absorption. This bistable nature is neverfound in vertebrate visual opsins of which photoproduct is unstable [1] and therefore
has been considered to contribute to the photoregeneration of opsins in photoreceptor
cells. In order to obtain a clue to understanding a contribution of the bistable nature to
photoreceptor cell responses in the wavelength discrimination, we established a mutant
zebrafish, which expresses UV-sensitive visual opsin instead of parapinopsin in thepineal photoreceptor cells.We performed calcium imaging for the pineal organ of themutant and wild-type zebrafish with a multiphoton microscope.

The result suggested

that the bistability of parapinopsin contributes to the pineal color opponency. In
addition, we discussed effect of parapinopsin bistability on photoreceptor sensitivity
under strong light conditions.
[1] Koyanagi, M.; Kawano, E.; Kinugawa, Y.; Oishi, T.; Shichida, Y.; Tamotsu, S.;
Terakita, A. PNAS 2004, 101, 6687-6691.
[2] Wada, S.; Kawano-Yamashita, E.; Koyanagi, M.; Terakita, A. PLoS One 2012,

e39003.
[3] Koyanagi, M.; Wada, S.; Kawano-Yamashita, E.; Hara, Y.; Kuraku, S.; Kosaka, S.;
Terakita, A. BMC biology 2015, 13, 73.

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Effects of micro-bubble assistance on the performanceof 3D integrated micro solution plasma
Reiya Nakagawa, Hiroto Masunaga, Yodai Ishida,
and Tatsuru Shirafuji
Department of Physical Electronics and Informatics,
Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka, 558-8585 Japan
1. Introduction
Plasma in liquid has attracted much attention because of their possible applications forsolving water-related environmental issues. We have previously proposed a novel
three-dimensionally integrated micro solution plasma (3D IMSP) reactor, which can generatea large number of microplasmas in a porous dielectric material filled with a gas/liquid mixed
medium [1]. However, 3D IMSP is not effective for the treatment of an aqueous solution witha high electrical conductivity [2], which was one of disadvantages of 3D IMSP. In this work,
we have introduced micro bubbles into the 3D IMSP reactor in order to treat water withhigher electrical conductivity.
2. Experimental procedure
We employed micro-bubble generator (Hack
UFB, FB11) as a substitute of the liquid
circulation pump used in our previous 3D IMSP
reactor [1], in which the circulation speed isfaster than in conventional 3D IMSP. The
aqueous solutions used for this experiment hadelectrical conductivities of 1, 10, 100, 200, 500,
and 1000 µS/cm, which were prepared bymixing KCl with deionized water andmethylene blue.

with
micro
bubbles
100 uS/cm

200 uS/cm

500 uS/cm

1 mS/cm

100 uS/cm

200 uS/cm

500 uS/cm

1 mS/cmwithout
micro
bubblesFig. 1 Effects of micro-bubble assistance on
the operation of 3D IMSP.

3. Results and discussion
As shown in Fig. 1, we can confirm that the
micro-bubble assisted 3D IMSP reactor can
generate plasma in the aqueous solution withelectrical conductivity up to 500 µS/cm, while
Fig. 2 Effects of micro-bubble assistance on theconventional 3D IMSP cannot. Furthermore,
ignition and sustain voltages of 3D IMSP.
micro-bubble assistance has an effect of
reducing the ignition and sustain voltages of 3D IMSP as shown in Fig. 2. Although theresults are not shown here, we have confirmed that micro-bubble assisted 3D IMSP candecompose methylene blue without reducing decomposition efficiency.
References:[1] T. Shirafuji and Y. Himeno, Jpn. J. Appl. Phys. 2013, 52, 11NE03.
[2] T. Shirafuji, J. Ueda, A. Nakamura, S.-P. Cho, N. Saito, and O. Takai, Jpn. J. App. Phys.
2013, 52, 126202.

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Template for the abstract

Effect of nitrogen codoping on strontium doped NaTaO3
Akiyo Ozawa1,2, Keita Kobayashi2, Muneaki Yamamoto3, Tomoko Yoshida4

1 Applied Chemistry and Bioengineering, Graduate School of Engineering, Osaka City
University, 3-313-8, Sugimoto, Sumiyoshi-ku, Osaka, Japan
2 Corporate Research Laboratories, Research & Development Division, Sakai Chemical
Industry, Co., Ltd., 5-1, Ebisujima-cho, Sakai-ku, Sakai, Japan
3 Materials, Physics and Energy Engineering, Graduate School of Engineering, Nagoya
University, Furo-cho, Chikusa-ku, Nagoya, Japan
4 Osaka City University Advanced Research Institute for Natural Science and Technology,
3-313-8, Sugimoto, Sumiyoshi-ku, Osaka, JapanFor potential applications in environmental protection and solar energy conversion,
heterogeneous photocatalytic reactions over semiconductor surfaces have been researched.
Among them, NaTaO3 having perovskite crystal structure was reported to provide highphotocatalytic activity under UV light irradiation. For example, Sr doped NaTaO3 modified by
Ag nanoparticles can reduce CO2 to CO with high efficiency [1]. However, the band gap of
NaTaO3 is too large to proceed photocatalytic reactions under visible-light irradiation. Most

solar energy is concentrated in the visible light region, therefore, from the perspective ofeffective utilization of sun light, many oxides/ complex oxides have been improved to use
visible light. Nitrogen doping of oxides or complex oxides has been reported to be aneffective method for allowing the use of visible light. However, high concentration nitrogen
doping into NaTaO3 by thermal treatment in ammonia atmosphere was rarely achieved [2].
Zhao et al reported that NaTaO3 doped with La can proceed to dope nitrogen, La-N co-doping
into NaTaO3 exhibited high activity for reduction of water in the presence of methanol

solution as a scavenger [3]. However, the role of cation doping in nitrogen doping was notwell understoodSr content in the sample. In this study, we synthesized nitrogen strontium
codoped NaTaO3 by solid state process and investigated effects of synthesis condition on thechemical state of doped nitrogen. In calcination at 1073 K in NH3, the Sr- N codoped sample

in which the Na site was substituted by 50 mol% of Sr contained 2 crystalline phase, NaTaO3and Sr2Ta2O7. On the other hand, in calcination at 1223 K in NH3, the sample maintained the
crystal structure of NaTaO3 even if containing 50 mol% Sr ion . As the amount of Sr doping
increased, Sr-N codoping into NaTaO3 inhibited generation of Ta3N5 in calcination under NH3
and enhanced nitrogen doping into NaTaO3.

References:[1] H. Nakanishi, K. Iizuka, T. Takayama, A. Iwase, A. Kudo, ChemSusChem 2017, 10, 112.
[2] X. Wang, G. Liu, Z. Chen, F. Li, G. Q. Lu, H. Cheng, Chem. Lett. 2009, 38 (3), 214.
[3] Z. Zhao, R. Li, Z. Li, Z. Zou, J. Phys. D: Appl. Phys. 2011, 44, 165401.

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Template for the abstract

Effects of Ga2O3 structure on the CO2 reduction with waterover Ga2O3 photocatalyst
Yu Kawaguchi1, Muneaki Yamamoto2, AkiyoOzawa1, Yuma Kato1,
Kokoro Yoshioka1, Tetsuo Tanabe3 and Tomoko Yoshida3
1
Graduate School of Engineering, Osaka City University, Osaka, Japan
2
Graduate School of Engineering, Nagoya University, Nagoya, Japan
3
Advanced Research Institute for Natural Science, Osaka City University, Osaka, Japan
Abstract: Gallium oxide photocatalyst (Ga2O3) has been the focus of attention, because it canconvert CO2 and water into CO, H2 and O2 under UV light irradiation. Recently, we revealed
that a Ga2O3 photocatalyst prepared by calcining a gallium nitrate powder at around 800 K
showed much higher activity for CO production than an Ag loaded β-phase Ga2O3, and theGa2O3 had the crystalline structure of mixed phases of γ- and β- Ga2O3 [1]. However, thereason for high activity of this Ga2O3 has not been revealed yet. In this study, we investigatedthat the reason for high activity for the Ga2O3 with mixed phases of γ- and β-phase.
Ga2O3 prepared by calcination of gallium nitrate powder for 4 h in air at 773 K. Fig.1 showstime course of CO production rate in the photocatalytic reduction of CO2 with water over the
Ga2O3, and Fig. 2 shows XRD patterns for this Ga2O3 before and after 1 h, 3 h and 5 hreaction test. As shown these figures, the photocatalytic activity of this Ga2O3 increased withtime, and GaOOH phase formed and grew in this Ga2O3. The Ga2O3 introduced GaOOH in
advance (Ga2O3-3wL) showed higher CO production activity than that of only Ga2O3 (Fig. 3),
indicating that the presence of GaOOH enhanced CO production. On the other hands, theactivity of single phase of GaOOH was low (shown in Fig. 3), suggesting the mixed phases ofγ, β-phase Ga2O3 and GaOOH is important to have high activity. FT-IR measurement revealed
that CO production would be enhanced with the increased CO2 adsorption on GaOOH phase.

Fig. 3 CO production rate for
Ga2O3, Ga2O3-3wL andcommercial
available
GaOOH (GaOOH-ca) after 5h reaction.
References: [1] Kawaguchi. Y.; Akatsuka. M.; Yamamoto. M.; Yoshioka. K.; Ozawa. A.; Kato.
Y.; Yoshida. T. J Photochem. Photobiol. A Chem. 2017.
Fig. 1 Time course of CO
production rate for Ga2O3.

68

Fig. 2 XRD patterns of
Ga2O3 (a) before andafter (b) 1 h, (c) 3 h and(d) 5 h reaction.

P-33

The Allosteric Receptor Containing a Zn-porphyrin anda Recognition Site for a Photo-responsive Stimulus Molecule
Chisato Okamoto,1 Yoshimitsu Tachi,1 Shuichi Suzuki,2 Keiji Okada,1,3
Masatoshi, Kozaki1,3
1Graduate School of Science, Osaka City University, 2Graduate School ofEngineering Science, Osaka University 3Osaka City University, Advanced Research Institutefor Natural Science and Technology (OCARINA)
Abstract: Artificial allosteric systems
with molecular recognition sites are
attracting much attention because of thepossible applicability to intelligent
catalysts and sensors.[1] Here, we
designed 1 and 2 as an allosteric receptor
and a photo-responsive stimulus
molecule, respectively. Receptor 1 has azinc diethynyldiphenylporphyrin unit
(DEDPP) as an active site and adiamidopyridine (DAP) and zinctetraphenylporphyrin terminals (TPP) as
molecular recognition sites. Stimulus
molecule 2 has both a pyridinyl (Py) anduracil terminals (U). Receptor 1 is able
to form a 1:1 complex with trans-2 by DAP-U multiple hydrogen bonds or a TPP-Py
coordination bond. A ligand is still accessible to DEDPP in this 1:1 complex (Active form).
On the other hand, cis-2 can form a 1:1 tight complex with receptor 1 by the concomitantformation of the coordination and the multiple hydrogen bonds. Consequently, a bulky
hexaphenylbenzene unit is fixed close to DEDPP and the ligand-binding ability of the zinc
porphyrin center is allosterically suppressed (Inactive form).
Allosteric receptor 1 was prepared in 32% yield by linking an alkyl chain 3 to a linear
conjugated chain 4 under the conditions of the Williamson ether synthesis. UV-vis spectra intoluene suggested that 2 reversibly isomerized using UV light (365 nm) and visible light (470nm). Both the formation of 1•cis-2 and the inhibition of ligand-binding ability of DEDPP
were monitored by UV-vis absorption measurement. The detailed synthesis and properties ofthese compounds will be presented.
Scheme 1. Synthesis of allosteric receptor 1.

[1] Sasaki, Y.; Suzuki, S.; Okada, K.; Kozaki, M. Tetrahedron Lett. 2016, 57, 4082–4085.

69

P-34
Template for the abstract

YgiT, antitoxin of mqsR-ygiT TA system, enhances
motility and biofilm formation in Escherichia coli
Ryoma Yokoi1 and Yoshihiro Yamaguchi1,21Department of Biology, Graduate school of Science, Osaka City University
2
The OCU Advanced Research Institute for Natural Science and Technology (OCARINA)
Abstract:
All free-living bacteria contain toxin or suicide genes, which regulate cell growthand cell death, reminiscent of the programmed cell death or apoptosis in mammaliancells. These toxin genes are co-transcribed with their cognate antitoxin genes andthus the systems are referred to as the toxin-antitoxin (TA) systems. Althoughprolonged induction of a toxin causes eventual cell death, it causes pseudo-dormancy
to the cells, in which all cellular metabolic function are maintained intact. Dormancy ofthe cells caused by TA toxins is considered to play important roles in biofilm formation.
Bacterial biofilms have a definitive role in medical device-associated diseases and also
mediate resistance to antibiotics and phagocytosis. MqsR is potent toxin encoded by
the mqsR-ygiT TA system in Escherichia coli. Although it has been reported that MqsR
causes an eight-fold induction of biofilm formation and that it also causes theactivation of a two-component system, the qseB-qseC operon, which is known to play
an important role in biofilm formation, MqsR has been shown to function as an mRNAinterferase specific for GCU sequences.
Here, we show that overproduction of YgiT antitoxin in the cell increases cell
motility and biofilm formation, indicating that YgiT antitoxin by itself is able to directly
act as a regulator for motility and biofilm formation (Fig. 1). We analyzed the amountof flagellin and curli fimbriae involved in cell motility. It was suggested that the overproduction of YgiT significantly repressed the expression of curli fimbriae but not
flagellin. Since YgiT itself can bind to the palindromic sequences in its own promoter
region similar to the MqsR-YgiT complex, it is possible that YgiT itself functions as aregulator binding to the promoter regions to repress or activate some genes leads toenhance cell motility and biofilm formation. We are screening the gene(s) enhancing
cell motility after induction of YgiT.

Fig. 1 : MqsR-YgiT
toxin-antitoxin system

70

P-35

Selective formate reduction catalyzed withaldehyde dehydrogenase using artificial co-enzyme
based on reduced form of methylviologen
Tomoya Ishibashi1, Shusaku Ikeyama2, Yutaka Amao1,2
1

Graduate School of Science, Osaka City University,

2

The Advanced Research Institute for Natural Science and Technology, Osaka City University

CO2 is reduced to methanol via formic acidand formaldehyde with the system of reduced
methylviologen (MV ･ +) as an artificial coenzyme, formate (FDH), aldehyde (AldDH)
and alcohol dehydrogenases (ADH) [1]. The
enzymatic kinetic parameters for FDH
catalyzing CO2 conversion to formic acidwith MV･+ or ADH catalyzing formaldehyde
conversion to methanol with reduced MV･+
have been clarified. However, the kinetic
parameters for AldDH catalyzing formic acid conversion to formaldehyde with MV･+ have notbeen clarified. In this study, the parameters for formic acid reduction with MV･+ and AldDH
were determined by enzymatic kinetic analysis.
MV ･+ (25~500 μM), sodium formate (2.0 mM) and AldDH (1.3 μM) in 1.0 mM sodium
pyrophosphate buffer (pH = 7.4) was reacted for 1 min at 30.5 ℃ in argon atmosphere. Sodium
dithionite was used as a reducing agent for MV. The decrease amount of formic acid as aformaldehyde production after 1 min was measured by ion chromatography. The decreaseamount of formic acid was plotted against the concentration of MV･+ as initial production rateof formaldehyde. Table 1 shows the kinetic parameters for FDH, AldDH and ADH with MV･+.
Comparing the kinetic parameters with FDH and ADH, it was found that the affinity between
MV･+ and AldDH is stronger than FDH and ADH.

References[1] Y. Amao, T. Watanabe, Chem. Lett., 2004, 33, 1544.
[2] S. Ikeyama, Osaka City University, 2017 Doctor’s thesis.
[3] R. Kataoka, Oita University, 2014 Master’s thesis.

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Light-Driven Water Oxidation Reaction Catalyzed by Cyano-Bridged
Metal Complexes with Core-Shell Structure
Akira Kitase, Hiroyasu Tabe, Yusuke Yamada
Graduate School of Engineering, Osaka City University
3-3-138 Sugimoto Sumiyoshi-ku, Osaka-shi, 558-8585, JAPAN
Abstract: Artificial photosynthesis is a promising method to convert chemical energy fromsolar energy. Artificial photosynthesis for water splitting includes several steps, 1) lightharvesting and chargeseparation, 2) water reduction and 3) water oxidation. In these steps,
water oxidation is the bottle neck to realize artificial photosynthesis, thus, a highly active
water oxidation catalyst (WOC) is strongly demanded. Recently, cyano-bridged metal
complexes, known as Prussian blue analogues, have been reported as efficient WOCs [1, 2].
We report herein the preparation of core-shell structured cyano-bridged metal complexes
having a catalytically active shell, CoII1.5[CoIII(CN)6], with various thickness and an inert core,
MNx[MC(CN)6] (MN = CuII, CoII; MC = FeIII, CoIII) to reduce the using amount of Co ion [3].
Light-driven water oxidation was examined under visible-light irradiation in a phosphatebuffer solution (pH 8.0) containing cyano-bridged metal complexes with core-shell structure
as a catalyst, [Ru(bpy)3]2+ as a photosensitizer and Na2S2O8 as a sacrificial oxidant.
CuII1.5[FeIII(CN)6]@CoII1.5[CoIII(CN)6] (190 nm core and 8 nm shell) exhibited catalytic
activity as high as CoII1.5[CoIII(CN)6] even though it contains small amount of Co ion.

Figure 1: Overall catalytic cycle of light-driven water oxidation (left) and TEM image of
CuII1.5[FeIII(CN)6]@CoII1.5[CoIII(CN)6] (right).
References:[1] Yamada, Y.; Oyama, K.; Gates, R.; Fukuzumi, S. Angew. Chem. Int. Ed. 2015, 54, 5613.
[2] Yamada, Y.; Oyama, K.; Suenobu, T.; Fukuzumi, S. Chem. Commun. 2017, 53, 3418.
[3] Li, C. H.; Peprah, M. K.; Asakura, D.; Meisel, M. W.; Okubo, M.; Talham, D. R. Chem.
Mater. 2015, 27, 1524.

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Template for the abstract

Title: Persister formation mechanism mediatedwith DNA binding protein, YjjJ
Name of author(s): Yuki Maeda(1), Yoshihiro Yamaguchi(1,2)
Affilication and address of author(s):1:Department of Biology, Graduate School of Science, Osaka City University;
2:The OCU Advanced Research Institute for Natural Science and Technology(OCARINA), Osaka City University
Abstract:
Reminiscent of eukaryotic apoptotic programmed cell death, bacteria also contain alarge number of suicide genes, which are in general co-expressed with their cognate antitoxic
genes. These systems called the toxin-antitoxin (TA) systems are associated with cellular
dormancy, and play major roles in persistent multi-drug tolerance of many human pathogens.
Especially the overexpression of HipA, the toxin in HipA-HipB TA system, induces persister
state in which the cells show high stress tolerance containing antibiotics treatment. We
showed that despite being a homologue of HipA, YjjJ has different cellular target(s) than thatof HipA. Helix-turn-helix DNA binding motif present in the N-terminal of YjjJ is consistent
with the data that YjjJ has DNA binding activity. It was also showed that HipB, the cognate
antitoxin of HipA, also acts as an antitoxin for YjjJ.
Here, we report functional characterization of YjjJ. We tested if yjjJ is involved inpersister formation. Although the deletion of hipA, hipBA or yjjJ gene has little effect on
persister formation, the deletion of yjjJ and hipA genes significantly increased persister cell
formation indicating that both hipA and yjjJ are important for multi-drug tolerance in E. coli.
We constructed various YjjJ mutants and found that YjjJ S200E and N-terminal truncatedmutant (N10) have no effect on cell growth. Since DNA binding motif presents in thetruncated region, we examined the DNA binding activity of these YjjJ mutants. Neither
S200E nor N10 truncated mutant binds to genomic DNA suggesting that YjjJ binds to DNA
and repress specific gene(s) expression resulting in cell growth arrest. As a step towards
identification of the target genes of YjjJ, we investigated the DNA binding specificities by anin vitro selection approach. We show that YjjJ is able to bind preferentially to specific DNA
sequences. The consensus sequences are CCCTATAGTGAGTCGTATTA (Seq1) and
GGATCCCCGCTGAGCA- ATAACTAGA (Seq2). Interestingly, Seq2 contains the CRISPR
repeat sequence which is involved in the bacterial defense system against phage. Then weanalyzed the effect of hipB-hipA and hipB-yjjJ TA systems on phage resistance in E. coli. The
deletion of yjjJ gene increased the phage sensitivity indicating that yjjJ may play an importantrole in phage defense mediated with CRISPR system.

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P-38

Photocatalytic Hydrogen Generation Systems
Inside Cross-Linked Crystals of Hen Egg White Lysozyme
Hikaru Takahashi,a Hiroyasu Tabe,a Satoshi Abe,b Takafumi Ueno,b
Yusuke Yamadaaa) Graduate school of Engineering, Osaka city University
3-3-138 Sugimoto Sumiyoshi-ku, Osaka-shi, 558-8585, JAPANb) School of Life Science and Technology, Tokyo Institute of Technology, 4259-B55,
Nagatsuta-cho, Midori-ku, Yokohama 226-8501, JAPAN
Abstract: Photocatalytic hydrogen evolution systems can be constructed by combining aphotosensitizer and a hydrogen evolution catalyst in the presence of an appropriate electron
donor. Cooperative immobilization of a photosensitizer and metal nanoparticles acting ashydrogen evolution catalysts on a mesoporous metal oxide can provide a heterogeneous
system, which is more robust than homogeneous systems. However, amorphous nature ofconventional mesoporous materials disturbs precise and periodical immobilization of catalytic
components [1]. In this study, cross-linked crystals of hen egg white lysozyme (CL-HEWL)
were employed as mesoporous supports to construct an efficient photocatalytic H2 generation
system. Rose Bengal (RB) and platinum nanoparticles (PtNPs), which act as a photosensitizer
and H2 generation catalysts, respectively, were immobilized inside CL-HEWL. Single-crystal
X-ray structure analyses of the CL-HEWL suggested that a coordination site of a Pt ion
locates in immediate proximity to potential adsorption sites for RB (Figure 1). Effective H2generation was achieved under visble light irradiation in the presence of
β-dihydronicotinamide adenine dinucleotide (NADH) acting as a sacrificial electron donor(Figure 2) [2].

Figure 1. Crystal structure of CL-HEWL
immobilizing RB and precursors of PtNPs.
The Pt atoms are represented by spheres. Blue
residues indicate potential adsorption sites forRB.

Figure 2. The overall photocatalytic cycle of H2generation.

References: [1] Yamada, Y.; Tadokoro, H.; Fukuzumi, S., RSC Adv. 2013, 3, 25677.
[2] Tabe, H.; Takahashi, H.; Shimoi, T.; Abe, S.; Ueno, T.; Yamada, Y., Appl. Catal. B., inpress (DOI: 10.1016/j.apcatb.2018.01.046).

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In-vitro reconstitution of light-harvesting complexesof a siphonous green alga, Codium fragile    
Yuki Isaji1, Rei Tohda4, Nami Yamano2, Tetsuko Nakaniwa4, Genji Kurisu4,5,
Ritsuko Fujii2,3
1
Faculty of Science, 2Graduate School of Science, 3OCARINA, Osaka City University, Osaka
558-8585, Japan; 4Institute for Protein Research, Osaka University, Osaka 565-0871, Japan;
5
CREST, JST, Saitama 332-0012, Japan
Abstract: Photosynthetic light-harvesting complexes plays the crucial role of absorbing
sunlight and transferring the excitation energy to photoreaction center [1]. A siphonous green
alga, Codium fragile has a unique light-harvesting complexes, siphonaxanthin-chlorophylla/b-binding protein (SCP) [2]. Siphonaxanthin bound to the SCP absorbs green region of lightwhich is abundant under the sea whereas it dissolved in organic solvent absorbs blue [2]. The
reason of the large bathochromic shift has been considered to the specific interaction between
siphonaxanthin and aminoacid residue, but it is not clarified yet. In-vitro reconstitution is oneof the powerful method to investigate the specific interaction for photosynthetic
light-harvesting complex which requires pigments for their folding process [3]. Therefore we
aim to perform in-vitro reconstitution of SCP in order to figure out the specific interaction.
In this study, we prepared the pigments from cultivated siphonous green alga, andperformed in-vitro reconstitution by using the inclusion body containing artificially expressed
SCP-protein [4] and the natural pigments following the protocol established for plantlight-harvesting complex [5] (See Fig.1). We will present our recent progress on the poster.

Figure 1. Concept of the Reconstitution of light-harvesting complex [5]References:[1] R. Croce and H. van Amerongen, Nature Chem. Biol. 10 2014, 492-501.
[2] Z.-X. Chu and J. M. Anderson, Biochem. Biophys. Acta 806 1985, 154-160.
[3] R. Croce, S. Weiss, R. Bassi, J. Biol. Chem. 274 1999, 29613–29623.
[4] T. Nakaniwa et al., unpublished result[5] A. Natali, et al., J. Vis. Exp. 92 2014, e51852.

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Investigation of amino acid residues involved in spectral tuningof butterfly long-wavelength-sensitive opsins
Tomoka Saito1, Mitsumasa Koyanagi1,2, Tomohiro Sugihara1, Kentaro Arikawa3,
Akihisa Terakita1,2
1Department of Biology and Geosciences, Graduate School of Science, Osaka City University,
Osaka, Japan; 2OCARINA, Osaka City University, Osaka, Japan; 3 Departmrnt of
Evolutionary Studies of Biosystems, The Graduate University for Advanced Studies(SOKENDAI), Kanagawa, Japan;
Absorption spectra of opsin-based pigments are tuned from UV to red region based oninteractions between the chromophore and surrounding amino acid residues. Both vertebrates
and invertebrates possess long-wavelength-sensitive (LWS) opsins, which underlie color
vision. It has been also known that LWS opsins have independently evolved in each lineage,
which implies the diverse mechanisms of spectral tuning. In vertebrate LWS opsins, thespectral tuning mechanisms have been well understood by spectroscopic analyses withrecombinant pigments and their mutants. However, the spectral tuning mechanisms of
invertebrate LWS opsins, such as insects, are largely unknown due to difficulty in obtaining
recombinant pigments. Here we successfully estimated the spectral sensitivity curves of two
kinds of butterfly LWS opsins by using our new method based on light-dependent change of
second messenger in opsin-expressing cultured cells [1], showing the wavelengths of theirmaximum sensitivities are at ~570nm and ~545nm, respectively. Then we analyzed a series ofchimeric and site-directed mutants to identify amino acid residues responsible for the spectral
tuning that accounts for the different absorption maxima. Based on the results, we discuss the
spectral tuning mechanisms of the butterfly LWS opsins, compared with those of vertebrateones.
[1] Sugihara, T.; Nagata, T.; Mason, B.; Koyanagi, M.; Terakita, A. PLoS One 2016, 11(8)

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Catalytic Activity of Thiocyanato-Bridged Polynuclear Metal
Complexes for Hydrolysis of Organophosphates    
Masaaki Matsushima, Chihiro Terashima, Hiroyasu Tabe, Yusuke Yamada
Graduate School of Engineering, Osaka City University
3-3-138 Sugimoto, Sumiyoshi-ku, Osaka-shi, 558-8585 JAPAN
Abstract: Organophosphates used for agricultural
pesticides are ranked as the 2nd cause of poisoning
accidents in Japan. Thus, the removal of organophosphates
has been strongly demanded. Recently, cyano-bridged
polynuclear metal complexes have been reported to exhibit
catalytic activity for organophosphate hydrolysis, in which
complexes having the low crystallinity exhibit higher
catalytic activity. We report herein catalysis of Figure 1. Partial structure ofthiocyanato-bridged polynuclear metal complexes thiocyanato-bridged(FeIIIn[MS(SCN)6]; MS = IrIII, RhIII, PtIV), which have polynuclear metal complexes.
nonlinear MS–S–C bonds beneficial to decrease thecrystallinity, for organophosphate hydrolysis. (Figure 1).
The precursor compounds, {(n-C4H9)4N}3[MS(SCN)6] (MS = IrIII and RhIII) andK2[PtIV(SCN)6], were synthesized according to the previous reports [1,2]. Then,FeIIIn[MS(SCN)6] (MS = IrIII, RhIII, PtIV) complexes were obtained as precipitates by mixing asolution of the corresponding complex and that of FeIII(NO3)3 under ambient conditions.
X-ray diffraction patterns of FeIIIn[MS(SCN)6] showed that FeIII2/3[PtIV(SCN)6] has the highcrystallinity while FeIII[IrIII(SCN)6] and FeIII[RhIII(SCN)6] have the low crystallinity.
Heterogeneous catalysis of each complex was evaluated by the hydrolysis of
4-nitrophenylphosphate disodium salt (NPP) in a buffer solution (pH 8.3) (Figure 2).
Turnover frequencies (TOFs) of FeIII[IrIII(SCN)6] and FeIII[RhIII(SCN)6] for NPP hydrolysis
were 0.44 and 0.41 h–1, respectively, higher than that of FeIII2/3[PtIV(SCN)6] (TOF = 0.24 h-1).

Figure 2. Catalytic hydrolysis of 4-nitrophenylphosphate disodium salt.
References:[1] Fricke, H. H.; Preetz, W. Z. Anorg. Allg. Chem. 1983, 507, 23.
[2] Siebel, E.; Fischer, R. D. Chem.–Eur. J. 1997, 3, 12.

77

P-42

Synthesis and Characterization of Li2Co1.8Ni0.2O4 as
Zero-Strain Lithium Insertion Material
Kensuke Kajikawa, Yusuke Yamada, Kingo Ariyoshi
Graduate School of Engineering, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka, 558-8585, Japan
Lithium-ion batteries are the key components for portable electronic devices, electricvehicles, and renewable energy storage systems. The performance of lithium-ion batteries
depends on lithium insertion materials used as positive and negative electrodes. Zero-strain
lithium insertion materials are beneficial to obtain long-life batteries, because no change in thelattice dimension during charge and discharge results in low mechanical stress. In this study,
we report the synthesis and characterization of Li2Co1.8Ni0.2O4 having spinel framework as anovel zero-strain lithium insertion material. Lithium cells with Li2Co1.8Ni0.2O4, which was
prepared by solid-state reaction at 450°C, showed reversible capacities of about 110 mA h/g atconstant operating voltage of 3.5 V, indicating that the lithium-insertion reaction of Li2Co1.8Ni0.2O4 proceeds in a two-phase manner as shown in Fig. 1. To examine change in the

Schematic illustration of lithium inserion/extraction reaction for Li2Co1.8Ni0.2O4.

78

10

20

*

*

30

40

50

60

2 / degreestrain material.
In the poster session, we discuss
the mechanism of “zero-strain”

Fig. 2

reaction of Li2Co1.8Ni0.2O4.

insertion/extraction reaction.

70

531

**

・
333

*

331

**

222

*

440

311

400

crystal structure of Li2Co1.8Ni0.2O4
during lithium insertion/extraction
reactions,X-raydiffraction
measurements of the electrodecharged to 4.2 V were carried out.
As shown in Fig. 2, no significant
change in the diffraction peaks
before and after charge evidenced
that Li2Co1.8Ni0.2O4 is the zero-* polyethylene
・ Al

111

Fig. 1

80

90

FeK

100

Powder X-ray diffraction patterns of Li-

2Co1.8Ni0.2O4 before (black) and after charge (red). Cubic

latticeconstantdid

notchangeduringlithium

P-43

Color opponency with a single kinds of bistable opsin in thezebrafish pineal organ.
Seiji Wada1, Baoguo Shen1, Emi Kawano-Yamashita1, Takashi Nagata1,
Masahiko Hibi2, Satoshi Tamotsu3, Mitsumasa Koyanagi1,4, Akihisa Terakita1,41Department of Biology and Geosciences, Graduate school of Science, Osaka City University,
Japan; 2Division of Biological Science, Graduate School of Science, Nagoya University,
Japan;3Department of Biological Sciences, Faculty of Science, Nara Women’s University,
Japan;4OCARINA, Osaka City University, Japan;
Color perception is achieved through antagonistic responses to wavelengths of light, i.e.,“color opponency”. Ocular and extraocular color opponency has been considered to require
multiple color-opsins. Human trichromacy involves red-, green- and blue-sensitive cone
opsins, which are exclusively present in different photoreceptor cells. Intriguingly, it wasreported that pineal organs of lower vertebrates discriminate UV and visible light; pineal
ganglion cells receiving the light information from several pineal photoreceptor cells exhibit
suppression and promotion of neural firings to UV and visible light, respectively. We havefound that parapinopsin is a UV-sensitive pigment and is possibly the common molecularbasis for UV-sensitivity in the color opponency of pineal related organs [1-3]. Interestingly,
parapinopsin has a molecular property called bistable nature; upon UV light-absorption, theUV-sensitive parapinopsin converts to a stable photoproduct that is sensitive to visible light,
and this photoproduct reverts to the original dark state upon visible light absorption, showinginterconvertibility between the dark state and its photoproduct. Therefore, parapinopsin has
two stable “color states”, unlike other vertebrate color opsins of which photoproduct isunstable. However, a contribution of the bistable nature providing the visible light-sensitive
photoproduct in the pineal color opponency has not been understood. In this study, we carriedout calcium imaging using parapinopsin-deficient zebrafish to obtain a clue to how thebistable nature of parapinopsin contributes to pineal color opponency. As a result, we foundthe pineal color opponency is achieved through the photo-interconversion of parapinopsin.
Based on obtained results, we discuss the mechanism of pineal color opponency with a single
kind of opsin, parapinopsin.
[1] Koyanagi M.; Kawano E,; Kinugawa Y.; Oishi T.; Shichida Y.; Tamotsu S.; Terakita A.
PNAS 2004, 101, 6687-6691[2] Wada S.; Kawano-Yamashita E.; Koyanagi M.;Terakita A. PLoS One 2012, e39003[3] Koyanagi M.; Wada S.; Kawano-Yamashita E.; Hara Y.; Kuraku S.; Kosaka S.;
Kawakami K.; Tamotsu S.; Tsukamoto H.; Shichida Y.; Terakita A. BMC Biology 2015, 13,
73.

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Study on the structure and CO2 reduction activityof Al2O3 supported Ga2O3 photocatalyst
photo

Ryota Ito1, Muneaki Yamamoto2, Akiyo Ozawa1, Yuma Kato1,
Yu Kawaguchi1,Masato Akatsuka2, Tetsuo Tanabe3 and Tomoko Yoshida3
1Graduate School of Engineering, Osaka City University, Osaka, Japan
Graduate School of Enjineering, Nagoya University, Nagoya, Japan
Advanced Reserch Institute for Natural Science, Osaka City Univercity, Osaka, Japan
Nowadays photocatalysts are expected to provide a solution for energy crisis and global
warming. In particular, photocatalytic CO2 reduction to give industrial usable products such as
CO gas has attracted much attention. It has been reported that Ga2O3 works as a photocatalytst
for CO2 reduction with water [1]. In order to improve the photocatalytic activity of Ga2O3, in
this study, Ga2O3 was loaded on Al2O3 support having high surface area. Preparing Al2O3
supported Ga2O3 (Ga2O3/Al2O3) samples with various amounts of loaded Ga2O3, theiractivities on photocatalytic CO2 reduction with water were examined, and the correlation of
the photocatalytic activity and the amount of loaded Ga2O3 and their structure wasinvestigated in detail.
Ga2O3/Al2O3 samples were prepared by impregnation of γ-phase Al2O3 with aqueous
solution of gallium nitrate followed by dry and calcination in air at 823 K for 4 h. The loading
amouts of Ga2O3 were 5, 10, 20, 40 and 60 wt%. Pure Al2O3 and Ga2O3 samples (refreed as 0
and 100 wt%, repectively) were also prepared in the similar procedure.
Fig.1 compares prodution rates of CO, which is the product of the photocatalytic CO2
reduction, for all prepared Ga2O3/Al2O3 samples. Among them, 40 wt% Ga2O3/Al2O3 showed
the highest CO production rate with CO selectivity similar to that of 100wt% Ga2O3. This
indicates that the activity of the photocatalytic CO2 reduction of Ga2O3 was enhanced with its
loading on Al2O3 support.
Fig.2 shows the difference XRD patterns for all prepared samples which were obtained bysubtracting the XRD pattern of Al2O3 support from that of each sample. Samples with loading
amounts of less than 20 wt% consisted of single α-phase Ga2O3, while 40 and 60 wt%
Ga2O3/Al2O3 consisted of α and γ phases. This suggests that existance of either α or γ phase or
coexistence of the both phase enhanced the photocatalytic activty. Understaning the role ofeach phase on the phtocatalytic activity remains for future work.

Fig.1 CO production rates for preparedsamples after 5 h reaction.

Fig.2 Difference XRD patterns of Ga2O3/Al2O3 andGa2O3 samples.

References:[1] L.Zhang et al.,108th Symposium of Catal. Soc. A,1D15 (2011).

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P-45

Effect of photon energy on photocatalitic CO2
reduction with water by Ag loaded Ga2O3
Kokoro Yoshioka1, Yu Kawaguchi2, Muneaki Yamamoto3,
Akiyo Ozawa2 Tetsuo Tanabe4 and Tomoko Yoshida4
1
Faculty of Engineering, Osaka City University, 2 Graduate School of Engineering,Osaka City University, 3Graduate School of Engineering, Nagoya University,
4 Advanced Research Institute for Natural Science, Osaka City University
Gallium oxide (Ga2O3) photocatalysts can promote CO2 reduction with water to produce
CO, H2 and O2 under UV photon irradaition. Recentry, it has been reported that Ag
loading on Ga2O3 as co-catalyst significantly improved the photocatalytic activity for CO2
reduction to CO[1]. In this study, we investigated the influence of photon energy on CO
production as the result of photocatalytic redcution of CO2 with water over Ag loaded
Ga2O3 (Ag/Ga2O3).
0.5 wt% Ag/Ga2O3 samples were prepared by impregnation (IMP) and photodeposition(PD) methods. In the photocatalytic reduction tests, the prepared samples were photoirradiated using a 300 W Xe lamp controling photon energy with band path or cut filters.
Fig. 1 shows energy spectra of photons after energy selection by filters (254bp, 33U and37L) together with energy spectrum of a Xe lamp (no filter). Fig. 2 shows diffuse
reflectance (DR) spectra of Ag/Ga2O3 samples after 5 h reaction by using photons filteredby 37L. The DR spectra showed a large absorption band at wavelength shorter than 290
nm, a small shoulder around 300 nm and a broad band around 450 nm. They wererespectively assigned to the band gap transition of the Ga2O3, the absorption of Ag small
clusters and a localized surface plasmon resonance (LSPR) of Ag nanoparticles. The
LSPR band was broadened to the longer wavelength for Ag/Ga2O3(PD) compared to thatof Ag/Ga2O3(IMP). This suggests that sizes of precipitated Ag nanoparticles were larger
on the former than that on the later [1].
In the reduction test by using photons filterd by 37L, both Ag/Ga2O3(IMP) and
Ag/Ga2O3(PD) were inactive. On the ther hand, by using photons filtered by 254bp, 33U
and no filters, the photocatalytic CO2 reduction proceeded over both samples, and
Ag/Ga2O3(IMP) showed higher CO selectivity. These results suggested that the band gap
excitation of Ga2O3 is necessary for the CO2 reduction, while the CO selectivity depends
on the size of Ag particles. Fig.3 compares the CO selectivity for Ag/Ga2O3(IMP) for
photons flitered by 33U and 254bp and non-filtered. Note that CO selectivity depended
on the photon energy giving the highest CO selectivity for the photons filtered by 33 U.

Fig. 1 Energy spectra of photons after
energy selection by filters together withenergy spectrum of a Xe lamp (no filter).

Fig. 2 DR spectra of Ag/Ga2O3
samples after 5 h reaction by using
photons filtered by 37L.

Fig. 3 The change of CO selectivity
for Ag/Ga2O3(IMP) by the kind of
the filter .

[1] N. Yamamoto, T. Yoshida, S. Yagi, Z. Like, T. Mizutani, S. Ogawa, H. Nameki and
H. Yoshida, e-J. Surf Sci. Nanotech., 2014, 12, 263

81

P-46

DNA-binding activity of the multifunctional enzyme,
nucleoside diphosphate kinase
Chinami Sako1, Masaaki Miyamoto2, Satoshi Akanuma3,
and Ryoji Masui1
1
Grad. Sch. Sci., Osaka City Univ., 2RFC, Kobe Univ.,
3
Facl. Hum. Sci., Waseda Univ.
Nucleoside diphosphate kinase (NDPK) is a phosphotransferase that catalyzes of the-phosphate from nucleoside triphosphate (NTP) to nucleoside diphosphate (NDP) and criticalfor maintaining the cellular NTP pools. Some eukaryotic NDPKs have DNA-binding
activity and are involved in various cellular functions such as metastasis suppression andtranscriptional regulation.
The DNA-binding activity of those NDPKs has beencharacterized biochemically, but their DNA-binding site and binding mode are still unclear.
In this study, we first examined the universality of the DNA-binding activity using sevendifferent NDPKs. As a result, three NDPKs including human NM23-H2 and reconstructed
ancestral NDPKs bound to DNA, whereas the other four including NDPK from
Schizosaccharomyces pombe and Thermus thermophilus did not. These results suggest that
DNA-binding activity is not universally shared among NDPKs. Human NM23-H2 had anability to bind to double-stranded DNA as well as single-stranded DNA including
G-quadruplex DNA. However, NM23-H2 did not bind to 15-mer/bp or less oligo DNAseven if those contained G-rich motifs. In some cases DNA binding was inhibited in thepresence of ATP. Replacement of the autophosphorylation site His with Ala abolishedinhibition of DNA binding by ATP. These results raise the possibility that DNA binds near
the nucleotide-binding site of NDPK. Further, replacement of two basic residues common to
NDPKs with DNA-binding activity (Arg42 and Lys135 of NM23-H2) with Ala decreased the
DNA-binding activity increased the inhibitory effect of ATP. These results suggest that
positively charged Arg42 and Lys135 are involved in DNA-binding activity. In the tertiary
structure of NM23-H2, these two residues are located at outer surface of the hexameric ringand near the subunit interface, suggesting that DNA binds to this region. NDPK with
DNA-binding activity bound to single-stranded DNA stronger than double-stranded DNA andalso to circular plasmid DNA. We also investigated the length the dependence and sequencespecificity of DNA binding. Based on these results, we discuss the DNA-binding sites in
NDPK.

82

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Characterization of arrestin binding to parapinopsin, aphotopigment involved in pineal wavelength discrimination
Takashi Nagata1, Mitsumasa Koyanagi1,2, Emi Kawano-Yamashita1,
Seiji Wada1, Akihisa Terakita1,2
1Department of Biology and Geosciences, Graduate School of Science, Osaka City University,
Osaka, Japan; 2OCARINA, Osaka City University, Osaka, Japan
Many lower vertebrates including fish have extraocular photosensitive organs, pineal andrelated organs, which are suggested to transmit spectral (color) information on ambient light,
especially the ratio of ultraviolet (UV) and visible light, to the brain. We have previously
reported that parapinopsin, a photosensitive protein opsin, is responsible for the UV reception
in the pineal organ of lampreys and also revealed some properties of parapinopsin such as its
photoreaction and the interaction with other proteins involved in intracellular signalingcascades [1-3]. Upon light absorption, parapinopsin converts to a thermally stable active statethat drives a cellular signaling cascade. This is in contrast to the property of visual opsins (i.e.
visual pigments), of which active states are not stable and lose activation ability of thesignaling cascade during the process of bleaching. Additionally, we previously found thatarrestins, which bind to active state of opsin to shut off the signaling, are different forparapinopsin and visual pigments; that is, parapinopsin and visual pigments bind to β-arrestin(non-visual arrestin) and visual arrestin, respectively [4]. In the current study, wecharacterized light-dependent binding of β-arrestin by parapinopsin in comparison with avisual pigment by live-cell assays using luciferase-based luminescent reporters. Our results
suggest that the molecular mechanism of parapinopsin for arrestin binding could be different
from that of visual pigments.

[1] Koyanagi, M.; Kawano, E.; Kinugawa, Y.; Oishi, T.; Shichida, Y.; Tamotsu, S.; Terakita, A.
Proc Natl Acad Sci U S A 2004, 101 (17), 6687-91[2] Tsukamoto, H.; Farrens, D. L.; Koyanagi, M.; Terakita, A. J Biol Chem 2009, 284 (31),20676-83[3] Kawano-Yamashita, E.; Koyanagi, M.; Wada, S.; Tsukamoto, H.; Nagata, T.; Terakita, A.
PLoS One 2015, 10 (10), e0141280[4] Kawano-Yamashita, E.; Koyanagi, M.; Shichida, Y.; Oishi, T.; Tamotsu, S.; Terakita, A.
PLoS One 2011, 6 (1), e16402.

83

P-48

Construction of Composite Photocatalysts for Water Oxidation Using
Silica Nanoparticles Assembly as a Mesoporous Support    
Gentato Sakamoto, Hiroyasu Tabe, Yusuke Yamada
Graduate School of Engineering, Osaka City University
3-3-138 Sugimoto Sumiyosi-ku, Osaka-shi, 558-8585, JAPAN
Abstract: H2 is regarded as a clean fuel to realize a sustainable society, because only water isemitted after burning. Photocatalytic water splitting is an ideal strategy to convert solar
energy to chemical energy, H2. A photocalytic water splitting system can be constructed by
combination of a photosensitizer, a water reduciton catalyst for H2 evolution, and a wateroxidation catalyst for O2 evolution. However, concomitant use of an H2-evolution catalysts
and water oxidation catalysts in areaction solution fails to produce H2,
because of the back electron transfer
between the catalysts. Suchundesired back electron transfer can
be avoided by co-immobilization of
Figure 1. Catalytic cycle of the photocatalytic water oxidation
the catalysts on a support. As wereportedpreviously,

a

novel

reaction.

photocatalytic H2-evolution system
was constructed utilizing theinterparticle
spacesofsilica-alumina
nanoparticles
assembly by immobilization of Pt
nanoparticles
andaphotosensitizer.[1] We report herein

Figure 2. Simultaneous immobilization of iridium hydroxide andthat concomitant immobilization of
PtNPs in the pore of silica nanoparticles assembly.
a water oxidation catalyst, iridiumhydroxide, and a photosensitizer,[Ru(2,2’-bipyridine)3]2+, in the interparticle spaces of silica-nanoparticles assembly, which
successfully evolve O2 from water under visible-light irradiation in the presence of asacrificial oxidizing agent, S2O82– (Fig.1). Then, co-immobilization of iridium hydroxide and
PtNPs in the interparticle spaces of silica-nanoparticles assembly was examined forphotocatalytic water-splitting (Fig.2).
Reference: [1] Yamada, Y.; Tadokoro, H.; Naqshbandi, M.; Canning, J.; Crossley, M.;
Suenobu, T.; Fukuzumi, S. ChemPlusChem 2016, 81, 521.

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Subcellular localization of internal structure in gliding
machinery of Mycoplasma mobile

Isil Tulum1,2 , Kenta Kimura1 and Makoto Miyata1,21) Graduate School of Science, Osaka City University, Osaka, Japan
2) OCARINA , Osaka City University, Osaka, Japan
Mycoplasmas are parasitic and occasionally commensal bacteria which lack peptidoglycan layer.
They represent the smallest self-replicating organisms and are fastidious nutritional requirements.
Mycoplasma mobile, a fish pathogen, exhibits a unique gliding motility with an average speed of
2.0 to 4.5 μm/s based on ATP hydrolysis. This motility has been shown not to be related to othermechanisms of bacterial motility, nor does it involve motor proteins known to be involved ineukaryotic cell motility. The gliding machinery is a complex composed of surface and internal
components. The internal structure consists of at least ten proteins and seven of them aretandemly coded with very short gaps on the genome. Two proteins, coded by the ORFs, MMOBs
1660 and 1670 are paralogs of F1-ATPase/synthase α and β subunits, respectively [1]. F1Fo
ATPases have been identified in most bacteria, including Mycoplasmas. Interestingly, besides theconventional F1Fo ATPase (Type 1), phylogenetic studies identified two types of F1-like ATPaseclusters, Type 2 and Type 3 which had emerged during mycoplasma evolution [2].
In this study, subcellular localization of α-subunits of Type 1, Type 2, and Type 3 F1-likeATPases and individual subunits of Type 2 F1-like ATPase subunit paralogs were visualized by
the previously developed EYFP tagging using total internal reflection fluorescence microscopy(TIRFM) [3]. Five out of six proteins of internal structure colocalized with surface gliding
structure, suggesting that internal structure is associated with the gliding machinery. Two of theType 2 F1-like ATPase transformants exhibited enhanced binding activity and one of thetransformants exhibited significantly decreased binding activity, suggesting these proteins may
involve in the gliding mechanism. The phylogenetic tree construction and sequence analyses
showed that gliding related Type 2 F1-like ATPase and surface gliding proteins also exist in
newly identified gliding Mycoplasmas, Mycoplasma agassizi and Mycoplasma testudineum(pathogens of desert tortoise). Although the proteins of internal structure were highly conserved,
the surface gliding proteins and 16S rDNA sequences were distinct, suggesting that the surface
structure differs according to the host surface while the internal gliding structure remains
conserved to generate the force.
References[1] Miyata M and Hamaguchi T. Current Opinion in Microbiology.2016, 29,15-21.
[2] Béven L, Charenton C, Dautant A, Bouyssou G, Labroussaa F, Sköllermo A, Persson A,
Blanchard A, and Sirand-Pugnet P. PLoS ONE. 2012, 7(6): e38793.
[3] Tulum I, Yabe M, Uenoyama A and Miyata M. Journal of Bacteriology. 2014, 196(10), 815.

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Motor Evolved from F-ATPase for Mycoplasma mobile Gliding
Takuma Toyonaga1, Takayuki Kato2, Akihiro Kawamoto2,
Noriyuki Kodera3, Toshio Ando3, Keiichi Namba2,4and Makoto Miyata1,5
1. Graduate School of Science, Osaka City University, Japan
2. Graduate School of Frontier Biosciences, Osaka University, Japan
3. Bio-AFM Frontier Research Center, Kanazawa University, Japan
4. OCARINA, Osaka City University, Japan
5. Quantitative Biology Center, Riken (QBiC), Japan
Mycoplasma mobile, a fish pathogenic bacterium, glides on solid surfaces with a uniquemechanism. The gliding machinery is divided into two parts: surface and internal structures.
The internal structure of the gliding machinery has 28 filaments, and each filament is apolymer of 17 particles made of paralogs of ATP synthase catalytic subunits [1, 2]. Thus theseparticles are suggested to be the motors for gliding. Previously, we visualized the 3D structureof the motor by electron cryomicroscopy and single particle image analysis. Two hexamers
similar to ATP synthase were paired by a frame and attached with eight arm-like extensions.
In the present study, we analyzed the image of internal filaments obtained bynegative-staining electron microscopy. The filaments were aligned at regular intervals withbridging structures, suggesting that the filaments form a sheet by the arms of the motor. The
sheet formation may contribute to the directionality and cooperativity of leg movements. To
clarify the movement of the motor, we have observed the motor by high-speed atomic force
microscopy.
[1] Miyata, M.; Hamaguchi, T. Curr. Opin. In Microbiol. 2016, 29, 15-21.
[2] Nakane, D.; Miyata, M. Proc. Natl. Acad. Sci. USA 2007, 104, 19518-19523.

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Template for the abstract

Gliding Behavior Analyses of Mycoplasma gallisepticum
Masaki MIZUTANI1, Makoto MIYATA1,2
1) Graduate School of Science, Osaka City University, Osaka, Japan
2) OCARINA, Osaka City University, Osaka, Japan
Mycoplasma gallisepticum, an avian pathogen, is closely related to human-pathogenic
mycoplasmas, Mycoplasma pneumoniae and Mycoplasma genitalium. It attaches to animal
tissue surfaces and glides to spread infected area. The gliding system is totally different fromthat of Mycoplasma mobile, well-studied mycoplasma gliding[1,2]. The gliding machinery in
Mycoplasma gallisepticum is composed of an internal core and surface adhesion proteins. The
surface adhesion proteins catch sialylated oligosaccharides (SOs) on host surface and propel
the cell forward using force generated by the internal core. In the present study, we analyzed
the gliding behavior in detail by using phase-contrast or dark-field optical microscopy.
Mycoplasma gallisepticum glided at a speed of 0.27 ± 0.09 µm/s on the KOH washed glass
coated with SOs and bovine serum albumin (BSA). To examine the gliding direction, we
measured the changes of cell axis for 5-s invertals. The averaged gliding direction relative tothe cell axis was 0.6 ± 44.6 degrees, showing that the gliding direction is not biased to the leftor right from the cell axis. In general, some gliding bacteria show the rolling of cell body ingliding[3,4]. To check whether Mycoplasma gallisepticum rotate or not in gliding, we attacheda 40-nm colloidal gold to a gliding cell and observed by dark-field microscopy. The cell andthe colloidal gold parallelly moved keeping constant intervals, indicating that the cell body is
not rolling in gliding movement. To investigate the relationship between cytoadhesion andgliding, we added 0 to 0.5 mM of free sialyllactose (SL), an SO, to gliding cells to decrease thenumber of binding adhesin. The gliding speed linearly decreased to 0.10 µm/s, but the gliding
cell ratio sigmoidally reduced to 1.8%, indicating drag caused by adhesins or the cell body. To
determine the direct energy source of gliding, we treated gliding cells with detergent topermeabilize the cell membrane, resulting in the removal of cytoplasm[5]. The cells treatedwith only detergent stopped gliding simaltaniously with permeabilization, but somepermeabilized cells treated with detergent containing ATP continued gliding at 0.02–0.03 μm/s
for at least 1 min, indicating that the direct energy source of their gliding is possibly ATP.
[1] Nakane, D.; Miyata, M. Journal of Bacteriology 2009, 191, 3256－3264[2] Indikova, I.; Vroka, M.; Szostak, M. P. Veterinary Research 2014, 45, 99[3] Morio, H.; Kasai, T.; Miyata, M. Journal of Bacteriology 2016, 198, 283－290[4] Nakane, D.; Sato, K.; Wada, H.; McBride, M. J.; Nakayama, K. 2013 PNAS 110, 11145－
11150[5] Uenoyama, A.; Miyata, M. 2005 PNAS 102, 12754－12758

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Structural changes of Gli123 protein,essential for Mycoplasma mobile gliding    
Daiki MATSUIKE1, Yuhei O. TAHARA1, 2, Tasuku HAMAGUCHI1, 2,
Munehito ARAI3, Makoto MIYATA1, 2
1) Department of Biology, Graduate School of Science, Osaka City University, Japan2) The OCU Advanced Research Institute for Natural Science and Technology (OCARINA),
Osaka City University, Japan
3) Graduate School of Arts and Science, The University of Tokyo, Japan
Abstract:
M. mobile, a fish pathogenic bacterium glides on solid surfaces based on ATP energy by aunique energy-conversion mechanism. Three huge proteins clustering on the surface of thegliding machinery are essential for this mechanism. Gli349 shaped like an eighth music note(♪), acts as a leg protein by binding to sialylated oligosaccharides on solid surfaces. Gli521
shaped an interrogation mark (?), transmits the force to Gli349 as a crank protein [1].
Gli123 is a 123 kDa protein responsible for the assembly of surface gliding proteins in the"neck" part on cell surface [2].
Previously, we reported that three-dimensional reconstitution of Gli123 shaped like a"mushroom" with dimensions, 20.0, 14.5, and 16.0 nm, based on negative-staining electron
microscopy (EM), which was common with recombinant Gli123 (rGli123) isolated from
Escherichia coli cells.
In the present study, we focus on the conformational change of the rGli123 according to theionic strength, which was detected by light scattering. Rotary-shadowing EM showed thatrGli123 forms globular and filamentous structures with the lengths of 19.7 ±3.0 nm and 47.9
± 9.4 nm under 300 mM and 200 mM ammonium acetate, respectively. Limited proteolysis
showed flexibility of N-terminal 450 amino acid residues in the whole 1132 residues,suggesting that the flexible part can be assigned to the N terminus including a tansmembranesegment. Now, we are analyzing the conformation of molecule on a cell using the limited
proteolysis and immunoblotting.
References:[1] Miyata, M.; Hamaguchi, T. Current Opinion in Microbiology. 2016, 29, 15-21.
[2] Uenoyama, A.; Miyata M. Journal of Bacteriology. 2005, 187, 5578-5584.

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Structural Basis for Regulation of Contractile Function by
Phosphorylation of Cardiac Troponin I and T: Distance
Measurements Using Dipolar ESR Spectroscopy
Toshiaki Arata1,2,3
K. Sakai3, H. Yamashita3, C. Zhao3, T. Somiya3, S. Takai3, S. Ueki4, and M. Miki5
1
Dept. Biol., Grad. Sch. Sci., and 2OCARINA, Osaka City Univ.; 3Dept. Biol. Sci., Grad. Sch.
Sci., Osaka Univ., Osaka; 4Tokushima Bunri Univ., Kagawa; 5Univ. Fukui, Fukui. Japan
Continuous wave (CW) [1] electron spin/paramagnetic resonance (ESR/EPR), andpulsed EPR methods PELDOR/DEER [2] and DQC [3], can determine a distribution of the
distance 0.8-8.0 nm between two spin labels (nitroxide radical) attached chemically to protein
by site-directed genetic mutagenesis (SDSL-EPR). We applied these methods to huge protein
complexes like motor proteins myosin in muscle fibers [4] and kinesin-microtubule [5] andclock protein KaiABC complex [6] and detect their functional structural changes at atomic
level. Muscle contraction is triggered by Ca2+. Using EPR measurements we detected large
Ca2+-induced structural changes between troponin (Tn) subunits [7] but unexpectedly small
structural change of tropomyosin (Tm) on the actin filament for switching-on and allowing
myosin to produce force [8]. Heart muscle beating is fine-tuned by phosphorylation ofcardiac troponin (cTn) I and T. Here we focussed on the N-extension of cTnI (NxTnI) andN-domain of cTnT (N-cTnT) whose structures are unkown. The CWEPR spectra from two
spin labels attached on i and i+4 residues of NxTnI of full length cTnI were well fitted bynarrow 0.9 nm (alpha-helix) and broad 1-2 nm distance distributions (unstable conformation).
Upon cTnC-cTnI formation, the 43/47 residues on the protein kinase C (PKC)
phosphorylation sites Ser42/44 in NxTnI exhibited a 0.9 nm-peak and completely stabilizedalpha-helix, and the 23/27 on the PKA sites Ser23/24, narrowed the distribution at 1.4 nm andthen stablized an extended conformation. We propose that these regions are the main
determinants of the phosphorylation-dependent binding of NxTnI to cTnC and that thebinding is quite dynamic. cTnT combines cTnI with Tm. We have determined the interspin
distance between N-cTnT and Tm, and found that upon PKC phosphorylation of C-domain of
cTnT (C-cTnT) in the thin filament, the residual mobility of spin label bound to the N-cTnT
was markedly enhanced and the distance was markedly increased from less than 0.8 to 1.5-2.0
nm. This suggested that the loosening of N-cTnT-Tm binding upon PKC phosphorylation of
C-cTnT reduces muscle force and ATP hydrolysis rate in the heart.
[1]Ueki, S. et al. Biochemistry 2005, 44, 411. [2]Nakamura, M. et al. JMB. 2005, 348, 127.
[3]Abe, J. et al. Appl. Mag. Reson. 2012, 42, 273. [4]Arata, T. J. Mol.Biol. 1990, 214, 471.
[5]Sugata, K. et al. JMB. 2009;Yasuda et al. BBRC 2014. [6]Ishii, K. et al. GenesCells 2014.
[7]Aihara, T. et al. JBC. 2010, 285,10671 [8]Ueda, K. et al. Biophys.J. 2011, 100; 2013, 105.

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Gliding Machinery of Mycoplasma mobile
Visualized by High-speed Atomic Force Microscopy
Kohei KOBAYASHI1, Noriyuki KODERA2,
Yuhei O TAHARA1,3, Takuma TOYONAGA1,
Taishi KASAI1, Toshio ANDO2, Makoto MIYATA1,3

3.

1. Graduate School of Science, Osaka City University, Japan
2. Bio-AFM Frontier Research Center, Kanazawa University, JapanThe OCU Advanced Research Institute for Natural Science and Technology (OCARINA),
Osaka City University, Japan

Mycoplasma mobile, a fish pathogenic bacterium forms a protrusion at a cell pole and glides
in the direction of the protrusion with a unique mechanism. In the mechanism, "legs" on a cell
repeatedly catch, pull, and release sialylated oligosaccharides on substrate surfaces, driven byforce generated through ATP hydrolysis on an internal novel motor[1].
In this study, we tried to visualize the alignment of gliding machinery in vivo by high-speed
atomic force microscopy (high-speed AFM)[2].
Mycoplasma cells were fixed onto a glass slide by crosslinking with glutaraldehyde[3]. The
cells were alive on the glass, because they restarted gliding after 2 hours incubation in a
growth medium. The quick-freeze-replica electron microscopy showed that cells were fixed
onto glass surfaces with keeping their shape tapered.
The structures of internal motor consistent with those reconstructed by cryo-electron
microscopy were observed on the cells fixed onto a glass slide by the high-speed AFM,
suggesting that we succeeded in visualizing the internal motors beneath the cell membrane.
We found that the alignment of internal motors are tilted from cell axis, which may cause thebias direction of gliding[5].
[1] Miyata M.; Hamaguchi T. Current Opinion in Microbiology 2016, 29, 15-21.
[2] Ando T. Biophysical Review 2017, 9, 421–429[3] Yamashita H.; Taoka A.; Uchihashi T.; Asano T.; Ando T.; Fukumori Y. Journal of
Molecular Biology, 2012, 422, 300-309[4] Nishikawa MS.; Nakane D.; Toyonaga T.; Kawamoto A.; Kato T.; Namba K.; Miyata M.
submitted[5] Morio H.; Kasai T.; Miyata M. Journal of Bacteriology 2016, 198, 283-290.

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Structure and function of six cytoskeletal proteins in
Spiroplasma swimming
Daichi TAKAHASHI1, Aya KODAMA1, Katsumi IMADA2,
Makoto MIYATA1,3
1. Dept. Bio., Grad. Sch. Sci., Osaka City Univ.
2. Dept. Macromol. Sci., Grad. Sch. Sci., Osaka Univ.
3. OCARINA, Osaka City Univ.
Abstract: Spiroplasma eriocheiris is a pathogenic bacterium that causes a serious
disease to Chinese mitten crab [1]. Spiroplasma eriocheiris belongs to class Mollicutes,
and lacks the peptidoglycan layer like Mycoplasma species [1]. It swims using a uniquemechanism unrelated to flagella or pili, in which kinks propagate along its helical body[2]. The helical body is thought to be formed by six cytoskeletal proteins including
Fibril, a Spiroplasma specific protein, and five MreB proteins (MreB1 ~ MreB5),
bacterial actin homologs [3,4]. To elucidate the structure of these proteins, we
individually expressed them as recombinant proteins with N-terminal His-tag in
Echerichia coli. The cells expressing MreB1, MreB2 or MreB5 showed elongated cellmorphology with a few times longer than the original strain, suggesting their ability topolymerize. Filamentous structures were observed in the cells lysed by sonication,
under electron microscopy. To study the monomeric structures, the cells expressed forthe proteins were lyzed and subjected to centrifugation. Fibril and MreB4 formed
insoluble aggregates but others were soluble. MreB3 and MreB5 were successfully
purified and crystallized. X-ray structural analysis of these proteins is underway.

References[1] Wang W et al. Int J Syst Evol Microbiol 2011, 61, 703.
[2] Schaevitz JW et al. Cell 2005, 122, 941.
[3] Liu P et al. Front Microbiol 2017, 8, 58.
[4] Kürner J et al. Science 2005, 307, 436

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Template for the abstract

Effect of MreB Depolymerization
on Helicity-Switching Swimming Motility
in Crustacean pathogen, Spiroplasma eriocheiris
Yuya SASAJIMA1, Isil TULUM1,2, Makoto MIYATA1,2
1) Graduate School of Science, Osaka City University, Japan
2) OCARINA, Osaka City University, Japan
Spiroplasma eriocheiris, belonging to the class Mollicutes like Mycoplasma species, is ahelical-shaped swimming bacterium without peptidoglycan layer, causing, trembling diseaseof crustacean [1,2,3]. This bacterium has a unique motility system in which kinks propagate
along the cell body, based on its special internal structure involving five MreBs, a bacterial
actin, and Fibril specific to genus Spiroplasma [3]. In this study, to elucidate the role of
MreBs in the swimming, we observed the cells treated with A22, a MreB polymerization
inhibitor. When the swimming cells were exposed to A22, they lost the helicity and kinkpropagation. However, the cells nonspecifically fixed on glass surface showed thetranslocation of the cell density to several spots on the cell, whose positional intervals
increased with the A22 concentration used. The complete cytoskeletal structure were observed,
consisting of a dumbbel and a helical ribbon, at the front and along the body, respectively, inthe negative staining electron microscopy of the cell extracted by Triton X-100. The cell
treated with A22 showed some bulges along the cell on EM grid, consistent with theobservation by optical microscopy. The stacks of filaments were observed in the bulges bydetergent treatment. No helicity was observed in the filaments left on the ribbon trace after
A22 treatment. We isolated Fibril, known as another major component of the ribbon structure.
It showed characteristic repeated structure but no helicity. These results suggest that MreBs
interacting with Fibril form a helical ribbon as the cytoskelton, resulting in the helicity of cell.
[1] Miyata M and Hamaguchi T. Frontiers in Microbiology. 2016, 7, 960.
[2] Terahara N, Tulum I, and Miyata M. Biochemical and Biophysical ResearchCommunication. 2017, 487, 488-93.
[3] Liu P, Zheng H, Meng Q, Terahara N, Gu W, Wang S, Zhao G, Nakane D, Wang W, andMiyata M. Frontiers in Microbiology. 2017, 8, 58.

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Template for the abstract

Approach to biological specimens and material observation
using Quick-Freeze and Deep-Etch (QFDE) replica microscopy
Yuhei O Tahara1,2, Makoto Miyata1,21)Graduate School of Science, Osaka City University, Japan
2)OCARINA, Osaka City University, Japan

Quick-freeze and deep-etch replica is an electron microscopy method in which specimen isfrozen and fixed in a moment by slamming on a cooled metal and observed with resolution of
nanometer order and submillisecond time resolution. We adopt this technology in
Grant-in-Aid for Scientific Research on Innovative Areas, Ministry of Education, Culture,
Sports, Science and Technology in 2012 - 2016. Thirty two researchers and students from19 research groups inside and outside Osaka City Universuty attempted various subjects listed
below in four categories with related techniques, based on our helps for 226 days in the fiscal
2017.
(1) Motility machinery
Gliding machinery of Mycoplasma, Pili of Termus thermophilus Cytokinesis of slime moldand fission yeast, Flagella of thermophilic bacteria and Escherichia coli(2) Infectious microorganisms
Peptidoglycan desruption of Bacillus subtilis, Pili of pathogenic E. coli, Surface structureof Klebsiella pneumoniae and Ureaplasma.
(3) Useful microorganisms
Membrane structures of Saccharomyces cerevisiae and Schizosaccharomyces pombe andspores.
(4) Soft Matter
Surface structure of antimicrobial material, Carbon nanoparticles, Carbocyanine,
Lanthanide complex.

T. thermophilus

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B. subtilis on moth-eye film

S. cerevisiae

P-59

Elucidation of the environmental factors influenced on the
optical properties of fucoxanthin and chlorophyll c bound to
the fucoxanthin chlorophyll a/c protein, FCP
	
  	
  	
  	
  	
 
Nami Yamano1, Tadashi Mizoguchi2, Ritsuko Fujii1,3
1Grad. Sch. Sci., Osaka City Univ., 2Grad. Sch. Life Sci., Ritsumeikan Univ.,
3
The OCU Advanced Res. Inst. for Natural Sci. and Tech., Osaka City Univ..
FCP (Fucoxanthin chlorophyll a/c binding protein) is the major light harvesting complex
found in most-prospered aquatic-photosynthetic organisms such as diatoms and brown algae.
Light-harvesting complexes in photosynthesis play the important role in regulating the lightenergy transferred to the photoreaction center: The decrease in luminal pH upon highirradiance of sunlight triggers structural changes of light-harvesting complexes to enhancequenching ability of excess excitation energy [1]. In the case of FCP, however, the molecularmechanism of the regulative function has not been clarified yet. Two kinds ofcharacteristic-antenna pigments, chlorophyll c (Chl c) and fucoxanthin (Fx), absorb
blue-green region of solar energy and transfer the excitation energy to chlorophyll a (Chl a)when bound to FCP. Chl c was reported to show the changes in optical properties uponchanges in pH and/or ionic strengths of aqueous alcohols [2], as presumed from its chemical
structure (see Fig. 1). Fx bound to FCP proteins shows a large bathochromic shift ofabsorption spectrum, but this shift is known to be labile during purification steps, suggestingthat specific interactions around red-shifted Fx are easily modified by several environmental
factors. Energetic heterogeneities of both Chl c and Fx, which is induced by theenvironmental factors, might allow FCP to possess flexible photosynthetic capacity in vivo.
In this study, we isolated FCP from the diatom,
C. calcitrans, and measured its absorption,
circular dichroism, and fluorescence-excitation
spectra as the detergent micelle in a series ofbuffers with various pH values. We tried toevaluate the changes in the energy levels of Chl cand Fx bound to the FCP upon changing thesolvent factors, such as pH, ionic strength, andviscosity.
[1] C. Büchel, J. Plant Physiol. 172 (2015) 62-75.
[2] S. W. Jeffrey, and K. Shibata, Biol. Bull.136(1969) 54-62

Fig.1 Chemical structures ofpigments bound to FCP

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Template for the abstract

Study on in vivo 3D Micro-Tomographic
Visualization of Vascular Plexuses
and Capillary Blood Flow
Using Optical Coherence Doppler Velocigraphy    
Daisuke FURUKAWA1, Naoya KUSUMOTO1, Souichi SAEKI1, Masatsugu SHIBA2
Susumu AOKI3, Takafumi ITO3, Yoshiaki NISHINO3, Yusuke HARA1,4
1.
Graduate School of Engineering, Mechanical & Physical Engineering, Osaka City
University. 3-3-138, Sugimoto, Sumiyoshi-ku, Osaka, 558-8585
2

Graduate School of Medicine, Department of Gastroenterology, Osaka City University
3
TOKOTAKAOKA Co., Ltd, Hamamatsu, Japan
4
SHISEIDO Research Center, Yokohama, Japan

Abstract:
The skin aging process, e.g. wrinkles and saggings, caused by not only aging but also
ultraviolet irradiation, could be related to the depression of metabolic function. The
microcirculation system should be an important guideline of skin care for the anti/smart-aging.
Rheological behavior of interstitial in epidermal and dermal tissue, including blood
micro-circulation, can vary skin mechanics in micro scale, i.e. visco-elasticity. Therefore, anin vivo quantitative measurement of capillary blood flow velocity is crucial to clarify theirproperties. The purpose of this study is to visualize the tomographic flow velocity of red
blood cell in capillaries below human epidermal skin using Optical Coherence Doppler
Velocigraphy, i.e. OCDV [1]. This is constructed on a low coherence interferometer [2],which is based on Hilbert transform and adjacent auto-correlation. In order to validate OCDV
system, this was in vivo applied to human cheek skin under the condition of control. As aresult of skin tomography obtained by OCDV, en face cross-sectional imaging of doppler
velocity was found to display networks of capillary blood vessels in upper dermal tissue
around hair follicles, as well as morphological skin structure. It was confirmed that capillary
vasculature and blood velocity can be visualized tomographically even in the upper
subpapillary layer. In summary, OCDV system could be quite useful for a micro-tomographic
imaging of blood flow velocity of capillary vessels inside skin.
References:[1] Daisuke Furukawa, et al., “Accuracy Evaluation on Tomographic Micro-visualization of
Flow Velocity using High Frequency Modulated Low Coherence Interferometer, J. JSEM,
Vol.17, Issue 1, (2017), pp.52-56.
[2] Joseph M. Schmitt, “Optical Coherence Tomography (OCT): A Review”, IEEE Journal on
Selected Topics in Quantum Electronics, Vol.5, No.4 (1999), pp.1134-1142.

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Template for the abstract

Function of light-driven water-splitting device immobilizing
photosystem II inside porous glass plate    
Yusuke, Ikeda1 Tomoyasu Noji2, Keisuke, Kawakami 2 Tetsuro, Jin3 Nobuo,
Kamiya2,1
1Science, Osaka City University, 2The OCU Advanced Research Institute for Natural
Science & Technology (OCARINA), Osaka City University, 3National Institute of Advanced
Industrial Science and Technology, Osaka, Japan
Abstract: Researches to construct artificial photosynthesis utilizing photosystem II (PSII)
have been attractive approaches because PSII obtains electrons from water via light-driven
charge separation reactions of the quantum efficiency of 90%. In order to realize such kind of
artificial photosynthesis, it is important to clarify a condition that PSII works efficiently andstably. We previously constructed a light-driven water-splitting device by immobilizing PSII
and an electron acceptor 2,6-dichloroindophenol (DCIP) inside inner pores in porous glass
plates (PGPs) with 50-nm pore-diameter.[1] Although the activity of PSII was maintained
after immobilization, there was a problem that the excitation of PSII was inhibited byabsorption of DCIP in the visible region. In this study, a quinone was used as an acceptor thathas no absorption in the visible region. The oxygen evolved from PSII inside PGP was
detected by an oxygen electrode. The activity of PSII was also measured by breaching rate ofquinone after irradiation. The reducing rate of quinone was higher than that of DCIP. The
turnover number (TON) of PSII-quinone/PGP50 system was 105 e-/PSII that is 10 times
higher than that of PSII-DCIP/PGP50 system. The external qutum eficnecty (EQE) of the
PSII/PGP50 system was reahed to 11%, corresponding to a half of natural leaf (Figure 1).
We will discuss a mechanism of the increasing efficiency.

Figure 1. Water-oxidiation efficency of leaf vs PSII-quinone/porous glass plate.
References:[1] Noji, T.; Kawakami, K.; Shen, J-R.; Dewa, T.; Nango, M.; Kamiya, N.; Itoh, S.; Jin, T.
Langmuir, 2016, 32, 7796

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