The 2014 OCARINA Annual International Meeting.

The 2014
OCARINA Annual International Meeting.

Date：3 th-4 th March, 2015

Venue：10thFloor, Osaka City Univ. Media Center

Date： 3th-4th March, 2015
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Date
4th March, 2015 (Wed.)

10:00 - 18:00

5th March, 2015 (Thu.) 10:00 - 14:45

Venue
10th Floor, Osaka City Univ. Media Center

Banquet
4th March, 2015 (Wed.) 18:30-20:30
10th Floor, Osaka City Univ. Media Center

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

Supported by
Key Project of Osaka City University
COI-T hub for the “Realization of next-generation hydrogen-energy
society”

Organizing committee•

Isamu Kinoshita (Director of OCARINA / Graduate School of Science, Professor)•

Nobuo Kamiya (Professor of OCARINA / Director of ReCAP / Graduate Schoolof Science)•

Yutaka Amao (Professor of OCARINA / Professor of ReCAP)•

Keisuke Kawakami (Special-Associate Professor of OCARINA)•

Hajime Masukawa (Special-Associate Professor of OCARINA)

Advisory Board•

Richard J. Cogdell (University of Glasgow, UK, FRS, Professor)•

Masakazu Anpo (Special Adviser of Osaka Prefecture University)

Plenary lecture (Presentation: 55 mins, Question & Answer: 5 mins)•

Licheng Sun (Department of Chemistry, KTH Royal Institute of Technology,
Stockholm, SE, Professor)

Invited speakers (Presentation: 25 mins, Question & Answer: 5 mins)•

Ryu Abe (Kyoto University, JP, Professor)•

Masaya Matsuoka (Osaka Prefecture University, JP, Professor)•

Tomoko Yoshida (Nagoya University, JP, Associate Professor)•

Shigeru Ikeda (Research Center for Solar Energy Chemistry, Osaka University, JP,
Associate Professor)•

Rei Narikawa (Shizuoka University, JP, Associate Professor)•

Masami Nakazawa (Osaka Prefecture University, JP, Assistant Professor)•

Yoshinori Tsuji (University of Tsukuba, JP, Assistant Professor)•

Eiji Yamashita (Fuji Chemical Industry Co., Ltd., General Manager/COI-T
Position, Project Leader, JP)Invited speakers in Osaka City University•

Hideki Hashimoto (OCARNA, Project Leader / Graduate of Science, Professor)•

Shigeyuki Minami (OCARINA, Special Professor)•

Eriko Nakadai (OCARINA, Special-Associate Professor)•

Yoshihiro Yamaguchi (OCARINA, Special-Associate Professor)•

Masakazu Hirotsu (Graduate School of Science, Associate Professor)•

Hajime Masukawa (OCARINA, Special-Associate Professor)•

Seiya Kobatake (Department of Applied Chemistry, Graduate School ofEngineering, Professor)•

Naoteru Shigekawa (Department of Applied Physics and Electronics, Graduate
School of Engineering, Professor)•

Masaki Nakao (Department of Urban Engineering, Graduate School ofEngineering, Special-Appointed Professor)•

Makoto Miyata (Department of Biology, Graduate School of Science, Professor /OCARINA)•

Akihisa Terakita (Department of Biology, Graduate School of Science, Professor)•

Kazunobu Sato (Department of Chemistry, Graduate School of Science, Professor)•

Shuichi Suzuki (Department of Chemistry, Graduate School of Science, Associate
Professor)

Programme
4th March, 2015 (Wed.) 10:00-18:00
09:30-10:00

Start accepting

10:00-10:05

Opening Remarks

10:05-10:15

Message from Vice-President of Osaka City University

10:15-11:15
11:15-11:45
11:45-12:45
12:45-13:15[Plenary lecture] Licheng Sun (Department of Chemistry, KTH
Royal Institute of Technology, Stockholm, SE, Professor)[COI-T] Masaya Matsuoka (Osaka Prefecture University, JP,
Professor)
Poster sessions & Lunch Break[COI-T] Masakazu Hirotsu (Graduate School of Science, Associate
Professor)

13:15-13:45

Yoshinori Tsuji (University of Tsukuba, JP, Assistant Professor)

13:45-14:15

Rei Narikawa (Sizuoka University, JP, Associate Professor)

14:15-14:30

Break

14:30-15:00

Shigeru Ikeda (Research Center for Solar Energy Chemistry, Osaka
University, JP, Associate Professor)

15:00-15:30

Eriko Nakadai (OCARINA, Special-Associate Professor)

15:30-16:00

Yoshihiro Yamaguchi (OCARINA, Special-Associate Professor)

16:00-16:15

Break

16:15-16:45

Hajime Masukawa (OCARINA, Special-Associate Professor)

16:45-17:15

Ryu Abe (Kyoto University, JP, Professor)

17:15-17:45
17:45-18:00
18:30-20:30

Tomoko Yoshida (Nagoya University, JP, Associate Professor)
Hideki Hashimoto (OCARNA, Project Leader / Graduate of
Science, Professor)
Banquet5th March, 2015 (Thu.)
10:00-10:20
10:20-10:40
10:40-11:00
11:00-11:15
11:15-11:35
11:35-11:55
11:55-12:15
12:15-12:35
12:35-13:35
13:35-14:05
14:05-14:35

10:00 - 14:50

Seiya Kobatake (Department of Applied Chemistry, Graduate
School of Engineering, Professor)
Naoteru

Shigekawa(DepartmentofApplied

Physics

andElectronics, Graduate School of Engineering, Professor)
Masaki Nakao (Department of Applied Chemistry, Graduate
School of Engineering, Special-Appointed Professor)
Break
Makoto Miyata (Department of Biology, Graduate School of
Science, Professor / OCARINA)
Akihisa Terakita (Department of Biology, Graduate School of
Science, Professor)
Kazunobu Sato (Department of Chemistry, Graduate School of
Science, Professor)
Shuichi Suzuki (Department of Chemistry, Graduate School of
Science, Associate Professor)
Poster sessions & Lunch Break
Masami Nakazawa (Osaka Prefecture University, JP, AssistantProfessor)
Shigeyuki Minami (OCARINA, Special Professor)

14:35-14:40
14:40-14:50[COI-T] Eiji Yamashita (Fuji Chemical Industry Co., Ltd., General
Manager/COI-T Position, Project Leader, JP)
Closing Remarks

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Invited Lectures

1

Wednesday, 4th March2

3

Artificial Photosynthesis—From Molecular Catalysts to
Photoelectrochemical Cells for Water Splitting
Licheng Sun1,2
1

Department of Chemistry, School of Chemical Science and Engineering, KTH RoyalInstitute of Technology, 10044 Stockholm, Sweden; 2State Key Lab of Fine Chemicals,Institute of Artificial Photosynthesis, KTH-DUT Joint Education and Research Center on
Molecular Devices, Dalian University of Technology (DUT), 116024 Dalian, China

Inspired by the active sites of respective OEC in Photosystem II and Hydrogenases, a series ofmolecular catalysts for water oxidation and hydrogen generation have been developed in ourgroups during the past years. These molecular catalysts are based on transition metal
complexes, such as Ru, Fe, Cu, Ni and Co. For the water oxidation catalysts, we usednegatively charged carboxylate ligands and found that catalysts using these ligands are highly
active towards water oxidation in homogeneous systems driven by chemical, electrochemical
photochemical methods. For example, our recently synthesized Ru(bda) complex has shown arecord high turnover frequency (TOF) of >1000 s-1 for water oxidation in pH 1 solution withCe(IV) driven. Some very exciting results in electrochemical water reduction to hydrogen byusing a self-assembled super-efficient cobalt(II) based molecular catalyst have also beenachieved with very low overpotential and high stability. This molecular catalyst issignificantly more efficient and easier accessible than related molybdenum and cobalt
catalysts reported in literature.
Towards total water splitting without using any sacrificial reagents, photoelectrochemicalcells for light driven total water splitting based on the molecular water oxidation catalysts andmolecular hydrogen generation catalysts together with nanostructured semiconductors (suchas n-type TiO2 or p-type NiO), as well as on soft interfaces have been demonstrated in ourgroups and will be presented during the conference.
References:[1]

L. Duan, F. Bozoglian, S. Mandal, B. Stewart, T. Privalov, A. Llobet, L. Sun, Nature
Chem. 2012, 4, 418.

[2]

L. Duan, C. M. Araujo, M. S. G. Ahlquist, L. Sun, PNAS 2012, 109, 15584-15588.

[3]

F. Li, Y. Jiang, F. Huang, B. Zhang, Y. Gao, L. Sun, Angew. Chem. Int. Ed. 2012, 51,
2417.

4

[4]

L. Wang, L. Duan, B. Stewart, M. Pu, J. Liu, T. Privalov, L. Sun, J. Am. Chem. Soc.
2012, 134, 18868.

[5]

X. Li, M. Wang, D. Zheng, K. Han, J. Dong, L. Sun, Energy Environ. Sci. 2012, 5,
8220.

[6]

Y. Jiang, F. Li, B. Zhang, X. Li, X. Wang, F. Huang, L. Sun, Angew. Chem. Int. Ed.
2013, 52, 3398.

[7]

Y. Gao, X. Ding, J. Liu, L. Wang, Z. Lu, L. Li, L. Sun, J. Am. Chem. Soc. 2013, 135,
4219.

[8]

N. Wang, M. Wang, Y. Wang, D. Zheng, H. Han, M. Ahlquist, L. Sun, J. Am. Chem.
Soc. 2013, 135, 13688.

[9]

L. Chen, M. Wang, K. Han, P. Zhang, F. Gloaguen, L. Sun, Energy Environ. Sci.
2014, 6, 329.

[10]

B. Zhang, F. Li, F. Yu, X. Wang, X. Zhou, Y. Jiang, L. Sun, ACS Catalysis 2014, 4,
804-809.

[11]

M. Hansen, F. Li, L. Sun, B. König, Chem. Sci. 2014, 5, 2683-2687.

[12]

L. Wang, L. Duan, Y. Wang, M. Ahlquist, L. Sun, Chem. Commun. 2014, 50,
12947-12950.

[13]

X. Ding, Y. Gao, L. Zhang, Z. Yu, J. Liu, L. Sun, ACS Catalysis 2014, 4,2347-2350.

[14]

Y. Yang, M. Wang, L. Xue, F. Zhang, L. Chen, M. Ahlquist, L. Sun, ChemSusChem
2014, 7, 2889-2897.

[15]
[16]
[17]
[18]

L. Zhang, Y. Gao, X. Ding, Z. Yu, L. Sun, ChemSusChem 2014, 7, 2801-2804.
P. Zhang, M. Wang, Y. Yang, T. Yao, L. Sun, Angew. Chem. Int. Ed. 2014,
54,13803-13807 (Hot Paper).
D. Zheng, N. Wang, M. Wang, S. Ding, C. Ma, M. Darensbourg, M. Hall, L. Sun, J.
Am. Chem. Soc. 2014, 136, 16817-16823.
Z. Yu, L. Sun, Energy Environ. Sci. 2015, DOI: 10.1039/C4EE03565H.

5

Development of carbon neutral H2 production systemusing visible light-responsive TiO2 thin film photocatalyst
Masaya Matsuoka*, Yu Horiuchi, Masato Takeuchi, Masakazu Anpo
Osaka Prefecture University, Department of Applied Chemistry
1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, JAPAN
Photocatalytic H2 productions have been considered a promising environmentally
harmonious energy production process. We have successfully developed visible
light-responsive TiO2 (Vis-TiO2) thin films by controlling the substrate temperatures using aradio frequency magnetron sputtering (RF-MS) deposition method. This Vis-TiO2 thin film
can decompose H2O stoichiometrically and separately into H2 and O2 under solar lightirradiation by using H-type reaction cell [1,2]. In the present work, we have applied
Vis-TiO2 thin film for the separate evolution of H2 from aqueous solutions of various biomass
derivatives by employing an H-type glass cell. Vis-TiO2 thin films were prepared by an
RF-MS deposition method using a calcined TiO2 plate as the source material and Ar gas as
the sputtering gas. The films were prepared on a Ti metal substrate at a substratetemperature of 873 K. Evaluation of the photocatalytic activity was carried out by
photoelectrochemical measurements and the evolution of pure H2 from an aqueous solution of
biomass derivatives. It was found that photocurrents increase after the addition of 10 vol%alcohols as biomass derivatives into the HClO4 aqueous solution, indicating that thephotooxidation of alcohols occurred in preference to that of H2O, that is, the charge
recombination was suppressed due to the hole scavenging effect of alcohols. Then the
Vis-TiO2 thin film was applied for the separate evolution of H2 from an aqueous solution of
methanol under visible light irradiation (λ > 450 nm). Here, Pt was deposited on the
opposite side of the Vis-TiO2 on the Ti foil substrate before the photocatalyst was mounted onan H-type glass cell, separating two aqueous solutions. By adding 10 vol% methanol to theaqueous solution at the TiO2 side, the evolution rate of H2 increased from 0.08 to 0.52 µ mol /h, indicating that the use of alcohol was effective in improving the H2 evolution rate. In thissystem, the CO2 is evolved as the result of alcohol oxidation. However, this unnecessary CO2
can be utilized for the plant factory which consumes large amount of CO2 to accelerate thephotosynthesis of green plants. A new attempt will be introduced to construct the clean andcarbon neutral solar H2 production system by the combination of solar H2 production systemfrom water involving biomass derivatives using Vis-TiO2 thin film and plant factory.
[1] Schneider, J.; Matsuoka, M.; Takeuchi, M.; Zhang, J.; Horiuchi, Y.; Anpo, M.;
Bahnemann, D. W. Chem. Rev. 2014, 114, 9919.
[2] Horiuchi, Y.; Takeuchi, M.; Matsuoka, M.; Anpo, M. Phys. Chem. Chem. Phys., 2013, 15,13243.

6

Development of Hydrogen-Evolving Molecular Catalysts byUsing First-Row Transition Metal Complexes    
Masakazu Hirotsu
Graduate School of Science, Osaka City University, Osaka, 558-8585, Japan
Hydrogen is a clean energy carrier, and hydrogen production from water by renewable energy
sources is an ideal way to replace consumption of fossil fuels. Development of efficient andeconomical catalysts for the conversion of protons to hydrogen is a crucial issue to resolve.
Therefore, the proton reduction catalysts based on the earth-abundant metals have recently
been investigated [1]. In this study, we focus on the hydrogen-evolving molecular catalysts byusing first-row transition metal complexes.
We recently reported diiron complexes of N,C,S-tridentate ligands (L),[{Fe(L)(CO)2}Fe(CO)3] (Figure 1, A1, A2), as biomimetic models of the active site of[FeFe]-hydrogenases [2]. These diiron complexes showed electrocatalytic ability for proton
reduction of acetic acid in acetonitrile. To reduce the overpotential we designed analogous
diiron complexes with an amino group as a proton acceptor. Water-soluble first-row transition
metal complexes with S and N donor atoms were also explored to develop the electrocatalysts
for proton reduction. Nickel(II) complexes of thiolate-polyamine ligands (Figure 1, B)
exhibited high activity for the electrocatalytic proton reduction in the controlled potential
electrolysis at −1.1 V vs Ag/AgCl in water (pH 4.7).
A photocatalytic hydrogen evolution system with a water-soluble complex catalyst wasdeveloped by using [Ru(bpy)3]Cl2 and ascorbic acid as a photosensitizer and a sacrificial
electron donor, respectively. A cobalt(III) complex of a Schiff base prepared from
2-pyridinecarboxaldehyde and 4-methylthiosemicarbazide (Figure 1, C) acted as a catalyst forthe photocatalytic hydrogen evolution in water.

R''
OC

CO

S
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Fe

OC
OC OC

N
1

R
A1

OC

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S
Fe

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OC
N
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R2
A2

S

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N

Ni

N

N

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N
N

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N

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Figure 1. Metal complexes used as catalysts for proton reduction.
[1] Thoi, V. S.; Sun, Y.; Long, J. R.; Chang, C. J. Chem. Soc. Rev. 2013, 42, 2388.
[2] Hirotsu, M.; Santo, K.; Hashimoto, H.; Kinoshita, I. Organometallics 2012, 31, 7548.

7

Unique features of photosynthetic carbon metabolism
in the coccolithophore, Emiliania huxleyi    
Yoshinori Tsuji
Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba,
Ibaraki, 305-8577 Japan
Abstract:
Eukaryotic phototrophs acquired their chloroplast through multiple endosymbiotic eventsand then they are distributed across different lineages [1]. Despite the wide diversity ofeukaryotic algae, studies on photosynthetic carbon metabolism has been performed mainly inprimary endosymbiotic organisms such as green algae and land plants. On the other hand,studies on secondary endosymbiotic algae, such as diatoms and coccolithophores, are quite
limited in spite of their evolutionary and ecological importance.
Coccolithophores (division Haptophyta) are unicellular calcifying algae and known as a key
player in the global carbon cycle since they fix inorganic carbon by photosynthesis andcalcification. Their cells are covered with calcareous scales (coccoliths) consisting of CaCO3
and acid polysaccharide (AP). Emiliania huxleyi, the most dominant coccolithophore, arecharacterized by unique photosynthetic products such as AP, β-glucan and long-chain
unsaturated ketones called alkenones. Amongst these products, both β-glucan and alkenones
have been proposed as major storage carbon compound in E. huxleyi although no quantitative
analysis of synthesis of these compounds have been performed. We recently obtained
quantitative data on carbon flow into various compounds, and found that fixed carbon is
distributed into alkenones as the major storage compound, but negligibly into β-glucan [2].
This low carbon flux into β-glucan is unique since glucans (either α- or β-glucan) function as
major storage compounds in most photosynthetic organisms. Our results suggested theoccurrence of a novel regulatory mechanism whereby carbon flow into β-glucan are
maintained at low level in E. huxleyi.
Recent studies on secondary endosymbiotic algae showed substantial differences in thecentral carbon metabolism between primary and secondary endosymbiotic algae [3-6]. Since
the model of carbon metabolism established in land plants and green algae is not applicablefor secondary algae, further studies are necessary for the understanding and industrial
application of diverse algal carbon metabolism.
References:[1] Reyes-Prieto, A.; Weber, A.P.M.; Bhattacharya, D. Annu. Rev. Genet. 2007, 41, 147[2] Tsuji, Y.; Yamazaki, M.; Suzuki, I.; Shiraiwa, Y. Mar. Biotechnol. in press[3] Kroth, P. G.; Chiovitti, A.; Gruber, A.; Martin-Jezequel, V.; Mock, T. et al. PLoS One
2008, 3, e1426.

8

[4] Tsuji, Y.; Suzuki, I.; Shiraiwa, Y. Plant Cell Physiol. 2009, 50, 318[5] Tsuji, Y.; Suzuki, I.; Shiraiwa, Y. Plant Cell Physiol. 2012, 53, 1043
[6] Read, B. A.; Kegel, J.; Klute, M. J.; Kuo, A.; Lefebvre, S. C. et al. Nature 2013, 499, 209s

9Trial to create various light switches that work inphotosynthetic organisms   Rei Narikawa1,2, Keiji Fushimi1, Gen Enomoto3,
Ni-Ni-Win3 and Masahiko Ikeuchi3
1
Facul. of Sci., Shizuoka Univ., Shizuoka 422-8529,Japan, 2PRESTO,
JST, Saitama 332-0012, Japan, 3Grad. of Sch. of Arts and Sci., Univ. ofTokyo, Tokyo 153-8902, Japan
Abstract:
Cyanobacteria possess highly-developed photoresponsive systems to acclimate tochanging light environments using various photoreceptors. Among them,cyanobacteriochromes (CBCRs) are linear tetrapyrrole-binding photoreceptors that are foundonly from cyanobacteria to date and diverse in the chromophore species they contain and theircolour tuning mechanisms. Some of them are known to regulate certain photo-acclimation
processes such as phototaxis and light-dependent cell aggregation [1,2]. Further, we recently
succeeded in determination of the crystal structures of the chromophore-binding domains of
the CBCRs [3]. Based on these insights, we are now trying to create light switches that workin the photosynthetic organisms. Light switching system is conceptually divided into threebuilding blocks: light-input module, signal transduction module and transcriptional or
enzymatic output module. Thus, developments of these tree modules are individuallyproceeded.
As the light-input module, we are trying to create far-red/green light switches,
because these light qualities are not used so much for photosynthesis. For this purpose, we
focused on a red/green-type CBCR subfamily for a developmental platform. The
red/green-type CBCRs covalently bind phycocyanobilin and show reversible photoconversion
between red- and green-absorbing forms [4,5]. From this red/green-type CBCR subfamily, we
recently discovered two novel CBCRs: red/blue-reversible and far-red/orange-reversible
CBCRs [6,7]. We will show a future roadmap for creating far-red/green light switches basedon these two CBCRs. As the signal transduction module, we are trying to create light-induced
cAMP-synthetic system. We developed various chimeric proteins consist of the CBCR
chromophore-binding domain and a catalytic domain of the adenylate cyclase. As a result, we
succeeded in regulating cAMP synthesis by light stimulation. Further trials to obtainhigh-performance cAMP-synthetic light switches are now proceeding. As the transcriptionalor enzymatic output module, we just started the study. Thus, we will introduce only strategy
and preliminary results. Finally, we will discuss about prospect to apply the light switchingsystems to the other organisms and techniques.

10References:[1] Narikawa, R.; Suzuki, F.; Yoshihara, S.; Higashi, S.; Watanabe, M.; Ikeuchi, M. Plant
Cell Physiol. 2011, 52, 2214.
[2] Enomoto, G.; Nomura, R.; Shimada, T.; Ni-Ni-Win; Narikawa, R.; Ikeuchi, M. J. Biol.
Chem. 2014, 289, 24801.
[3] Narikawa, R.; Ishizuka, T.; Muraki, N.; Shiba, T.; Kurisu, G.; Ikeuchi, M. Proc. Natl.
Acad. Sci. U. S. A. 2013, 110, 918.
[4] Narikawa, R.; Fukushima, Y.; Ishizuka, T.; Itoh, S.; Ikeuchi, M. J. Mol. Biol. 2008, 380,
844.
[5] Fukushima, Y.; Iwaki, M.; Narikawa, R.; Ikeuchi, M.; Tomita, Y.; Itoh, S. Biochemistry
2011, 50, 6328.
[6] Narikawa, R.; Enomoto, G.; Ni-Ni-Win; Fushimi, K.; Ikeuchi, M. Biochemistry 2014, 53,5051.
[7] Narikawa, R.; Nakajima, T.; Aono, Y.; Fushimi, K.; Enomoto, G.; Ni-Ni-Win; Itoh, S.;
Sato, M.; Ikeuchi, M. Sci. Rep. 2015, 5, 7950.

11

Photoelectrochemical Water Splitting
Using Modified Chalcogenide Phtocathodes    
Shigeru Ikeda
Research Center for Solar Energy Chemistry, Osaka University,
1-3 Machikaneyama, Toyonaka, 560-8531, Japan
Solar water splitting for production of hydrogen (H2) energy has attracted much attention todate due to its environmental friendliness and cost-effective property. Since the first report of
PEC water splitting over a single crystalline TiO2 photoelectrode, various semiconductor
electrodes have been investigated for this purpose. Meanwhile, Cu-based p-typechalcogenides such as Cu(In,Ga)(S,Se)2 and Cu2ZnSn(S,Se)4 are widely used as
photoabsorbers in thin film solar cells. Due to their high absorption coefficients, tunable bandgap values (1.0-2.4 eV) by changing In/Ga and/or Se/S ratios, and suitable band alignment forwater reduction, these materials are also attractive for use as photocathodes for H2 production
from water.
The insertion of n-type thin layers on photocathodes to improve their performance of
PEC water splitting has been studied by several researchers using various p-type materials.
This modification is widely employed to facilitate efficient charge separation due to themodulated built-in field at solid-solid interface by the formed p-n junction. For the Cu-based
chalcogenide system, surface modification with an n-type CdS layer using a chemical bathdeposition (CBD) technique was typically employed; this resulted in improvement ofphotocurrent as well as onset potential compared to those of unmodified films.1 However, due
to the high toxity of CdS, replacement with alternative environment-friendly n-type layers
would be advantageous for practical use. Degradation and/or corrosion of used photocathodeare other critical problems of these CdS-modified photocathode system.
In the present study, we investigated surface modification of a Cu-based chalcogenide
thin film, CuInS2 with a novel n-type surface modifier, In2S3, for PEC water splitting.2 In2S3
possesses an indirect gap characteristic: it exhibits favorable transparency over the solar
spectrum due to the weak absorption in the blue region, despite its relatively narrow band gap(2.1 eV) compared to that of CdS. We show that the In2S3-modified CuInS2 worked morestablely and efficiently than the CdS-modified one as a photocathode for PEC water splitting.
References:[1] a) Ikeda, S; Nakamura, T.; Lee, S. M.; Yagi, T.; Harada, T.; Minegishi, T.; Matsumura, M.
ChemSusChem 2011, 4, 262. b) Ikeda, S.; Nonogaki, M.; Septina, W.; Gunawan; Harada, T.;
Matsumura, M. Catal. Sci. Tecnol. 2013, 3, 1849.
[2] Gunawan, Septina, W.; Ikeda, S.; Harada, T.; Minegishi, T.; Domen, K.; Matsumura, M.
Chem. Commun. 2014, 50, 8941.

12

Approaches to sensory processing mechanisms using
Caenorhabditis elegans models    
Eriko Kage-Nakadai
The OCU Advanced Research Institute for Natural Science and Technology, 3-3-138,
Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan
Abstract: Detection and processing of cues associated with food, poison or danger is crucial
for animals including humans. Molecular and cellular mechanisms underlying recognition andprocessing such stimuli remain largely unclear. We identified MBR-1 as a Caenorhabditis
elegans (C. elegans) ortholog of Mblk-1, a transcription factor that is expressed preferentiallyin the mushroom bodies of the honeybee brain. mbr-1 was expressed in restricted sets ofinterneurons in the head ganglia including AIM and RIF neurons. In the mbr-1 mutant adults,
an extra neurite connection between left and right AIM neurons, which should be eliminatedby the adult stage, were observed [1]. Wild-type and mbr-1 mutants are both normaly
attracted to the odorant benzaldehyde, while the mutant showed a defective ‘olfactorylearning’. Rescue experiments revealed that RIF, not AIM, was important for the olfactorylearning. Further analyses revealed that extra connection between the left and right RIF
neurons was not removed in mbr-1 mutants. We also show that Wnt pathway negatively
regulates the development elimination of extra connections in AIM and RIF interneurons [2].
We have recently purseued genetic approaches to address glia-neuron interaction in the
chemotaxis and avoidance behavior. RNAi screens by using glial marker transgenic animals[3] allowed us to identified ceh-26, which encodes C. elegans homolog of Drosophilaprospero / mammalian prox1, as a positive regulator of the glial cell differentiation andfunction. RNAi knockdouwn of ceh-26 caused the defects in the chemotaxis and avoidance
behavior. ceh-26 was selectively expressed in the glia, the excretory canal cell, andunidentified neurons. Because of the lethality of the ceh-26 deletion mutants, we developed a
conditional knockout (cKO) system based on the combination of the UV/TMP methods andpre-existing deletion mutants to generate ceh-26 cKO mutant strains [4-6].
References:[1] Kage, E.; Hayashi, Y. et al. Curr. Biol. 2005, 15, 1554-1559.
[2] Hayashi, Y.; Hirotsu, T.; Iwata, R.; Kage-Nakadai, E. et al. Nat. Neurosci. 2009, 12,981-987.
[3] Kage-Nakadai, E. et al. Biochem. Biophys. Res. Commun. 2011, 410, 471-477.
[4] Kage-Nakadai, E. et al. BMC Biothechnol. 2012, 12, 1.
[5] Kage-Nakadai, E. et al. Methods. 2014, 68, 397-402.
[6] Kage-Nakadai, E. at al. PLoS One. 2014, 9, e114680.

13

A Novel Mechanism for mRNA Interference by
Sequence-specific mRNA Interferases
Yoshihiro Yamaguchi
The OCU Advanced Research Institute for Natural Science & Technology(OCARINA), 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585 JAPAN.

Abstract: mRNA interference by antisense RNA in nature was described for the first time in1984 [1] and subsequently its principle was applied to regulate genomic gene expression [2]and also to create an immune system against viral infection called the mic-immune system [3].
We now found another novel mechanism in nature for mRNA interference by virtue of specificsequence-specific endoribonucleases called mRNA interferases [4]. By now, a large number ofhomologues to MazF mRNA interferases have been discovered, which cleave specific RNA
sequences from three to five bases. Here we report that there is another MazF homologue(MazF-hw) specifically cleaving a 7-base RNA sequence and also that there is a new type ofmRNA interferase (MqsR) not homologous to MazF in the E. coli toxin-antitoxin system.
Haloquodratum walsbyl, an extreme halophilic archaeon living in salt-saturated aquatic
ecosystems contains a gene encoding a protein (called MazF-hw) homologous to E. coli
MazF-ec (ACA-specific). Induction of MazF-hw in E. coli resulted in complete cell growtharrest only after 5 generations in contrast to almost immediate growth arrest by MazF-ec
induction. Purified MazF-hw cleaved only at a single site in 3.5kb MS2 phage RNA, but was
unable to cleave E. coli 16S and 23S rRNAs and yeast 18S and 26S rRNAs. Determination of
the cleavage site in MS2 RNA and following computer analysis of all these RNAs revealed
that MazF-hw cleaves at either UACUCA or UUACUC. Subsequently, using synthetic
oligoRNAs containing either one of the sequences, MazF-hw recognition sequence wasdetermined to be UUACUCA. This sequence was found to be unusually abundant in the genes
for ABC transporters, suggesting that MazF-hw may play an important role in adaptation ofthis archaeon to the extremely harsh ecosystems [5].
The mqsR gene has been shown to be positively regulated by the quorum sensing
auto-inducer, AI-2, which in turn activates a two-component system, the qseBqseC operon,playing an important role in biofilm formation in Escherichia coli. However, its cellularfunction has remained unknown. Here, we found that one base downstream of mqsR, there is agene, ygiT, which is co-transcribed with mqsR. Induction of msqR caused cell growth arrest,
while ygiT co-induction recovered cell growth. We further demonstrated that MqsR (98 aminoacid residues) is a potent inhibitor of protein synthesis by degrading cellular mRNAs. In vivo
and in vitro primer extension experiments showed that MqsR is an mRNA interferase
specifically cleaving mRNAs at GCU. However, this mRNA interferase has no homology (less14than 10%) with MazF, thus represents a previously unknown new category ofribosome-independent mRNA interferases. The mRNA interferase activity of purified MqsR
was inhibited by purified YgiT (131 residues). MqsR forms a stable two-to-one complex with
YgiT, and the complex specifically binds to a 4-base palindromic sequence existing in the
promoter region of the mqsR-ygiT operon as well as mazEF [6]. These results suggest thatmRNA interference by mRNA interferases appears to be a widely-used mechanism in bacteria
and archaea for gene regulation under stress conditions and for regulation of pathogenicity.
mRNA interferases will open a new avenue for their numerous applications in biotechnology
and medical science such as gene therapy.
References:[1]
Mizuno, T.; Chou, M. Y.; Inouye, M. PNAS 1984, 81, 1966.
[2]
Coleman, J.; Green, P. J.; Inouye, M. Cell 1984, 37, 429.
[3]
Coleman, J.; Hirashima, A.; Inokuchi, Y.; Green, P. J.; Inouye, M. Nature 1985, 315,
601.
[4]
Zhang, Y.; Zhang, J.; Hoeflich, K. P.; Ikura, M.; Qing, G.; Inouye, M. Mol. Cell
2003, 12, 913.
[5]
Yamaguchi, Y.; Nariya, H.; Park, J.-H.; Inouye, M. Nat Commun 2012, 3, 607.
[6]
Yamaguchi, Y.; Park, J.-H.; Inouye, M. J. Biol. Chem. 2009, 284, 28746.

15

Genetic engineering of heterocyst-forming cyanobacterium
Anabaena sp. PCC 7120 to increase
photobiological hydrogen production
Hajime Masukawa
OCARINA, Osaka City University,
3-3-138 Sugimoto, Sumiyoshi-ku, Osaka, 558-8585, Japan
Abstract: Cyanobacteria are able to photobiologically produce hydrogen with water as thesource of electrons to reduce protons. The hydrogen gas is produced as the inevitableby-product of nitrogen fixation by nitrogenase. To reconcile two incompatible processes,
oxygenic photosynthesis and oxygen-sensitive nitrogenase reaction, approximately one cell of
10-20 vegetative cells along the filaments develops into differentiated cells called heterocyststhat provide a microaerobic environment, allowing the nitrogenase to function in an aerobic
environment.
In an efforts to improve the H2 production by this strain, various mutant strains have been
constructed by genetic engineering [1]: (i) inactivation of uptake hydrogenase whichreabsorbs H2 produced by nitrogenase [2]; (ii) site-directed mutagenesis of selected aminoacid residues near the active site of nitrogenase to redirect electron flow more selectively
toward proton reduction and away from N2 reduction [3], and (iii) random mutagenesis of
hetR, encoding the master regulator of heterocyst differentiation, to increase heterocyst
frequency. In several nitrogenase variants, H2 production activities was not decreased by thepresence of high concentrations of N2 [3] and they continuously produced H2 over 21 days [4].
Some hetR variants formed heterocysts at frequencies significantly higher than those for theparental strain. The technical challenge is to improve the energy conversion efficiency
attained under outdoor conditions over a relatively long period.
References:[1] Sakurai, H.; Masukawa, H.; Kitashima, M.; Inoue, K. J Photoch Photobio C 2013, 17,
1-25.
[2] Masukawa, H.; Mochimaru, M.; Sakurai, H. Appl Microbiol Biot 2002, 58 (5), 618-624.
[3] Masukawa, H.; Inoue, K.; Sakurai, H.; Wolk, C. P.; Hausinger, R. P. Appl Environ
Microbiol 2010, 76 (20), 6741-50.
[4] Masukawa, H.; Sakurai, H.; Hausinger, R. P.; Inoue, K. Int J Hydrogen Energ 2014, 39(34), 19444-19451.

16

Visible-Light-Responsive Photocatalysts
for Solar Hydrogen Production    
Ryu AbeGraduate School of Engineering, Kyoto University, Kyoto, Japan
Photocatalytic water splitting into H2 and O2 using a semiconductor photocatalyst hasreceived much attention recently due to the potential of this method for the clean productionof H2 from water utilizing solar energy.[1] Although a number of metal oxides have beenreported to be active photocatalysts for the water-splitting reaction, most only function under
UV light owing to the large band gap energy of the materials. Since almost half of allincident solar energy at the Earth’s surface falls in the visible region, the efficient utilizationof visible light remains indispensable for realizing practical H2 production. However, it had
been noted that it is intrinsically difficult to develop an oxide semiconductor photocatalyst
that has both a sufficiently negative conduction band for H2 production and a sufficiently
narrow band gap (i.e., <3.0 eV) for visible light absorption because of the highly positive
valence band (at ca. +3.0 V vs. NHE) formed by the O 2p orbital. Although some non-oxide
semiconductors, such as (oxy)sulfides, (oxy)nitrides, and organic dyes (including metal
complexes) posses appropriate band levels for water splitting under visible light, they are
generally unstable and readily become deactivated through photocorrosion or self-oxidation,rather than evolving O2.
In order to split water under visible
light, we have developed new type ofwater splitting system based on two-step
photoexcitation between two different
photocatalyst materials [2], as shown in
Fig. 1. Over a H2 evolution photocatalyst,
the photoexcited electrons reduce water to
H2 and holes oxidize a reductant (Red) toan oxidant (Ox). The Ox is reduced backFig. 1 Photocatalytic water splitting under visible lightto the Red by photoexcited electrons
through two-step photoexcitation.
generated
over
an
O2
evolution
photocatalyst, where the holes oxidize water to O2. Based on the strategy, we have achieved
overall water splitting using various visible light responsive photocatalysts, such as SrTiO3
doped with Cr,[3] tantalum oxynitrides (TaON or BaTaO2N),[4] and organic dyes,[5] which
work as a H2 evolution photocatalyst, combined with WO3 for O2 evolution in the presence ofa shuttle redox mediator such as iodate/iodide (IO3–/I–). The use of BaTaO2N or coumarinorganic dye was demonstrating to be photoactive at wavelength up to ca. 700 nm, indicating
the potential of a two-step water-splitting system for utilizing a broader band of visible

17

spectrum. The introduction of two-step photoexcitation mechanism (Z-scheme) also enabled
us to generate H2 separately from O2, which will be quite important in the practicalapplication to avoid the explosion of the mixture, by employing simple porous glass filtersthat divide the two different photocatalysts but permit the transportation of redox couple.
Some metal oxynitride materials possess appropriate band levels for water splitting aswell as a narrow band gap that permits visible light absorption; are thus a promising candidate
as photoanode materials for efficient water splitting under visible light with low applied bias.
However, the introduction of N 2p orbitals in the valence band generates a new problem instability that need to be solved. Most oxynitrides undergo self-oxidative deactivation somedegree, in which photogenerated holes oxidize nitrogen anions (N3–) to N2 (2N3– + 6h+ → N2).
Recently, we have developed a simple method for
fabricating stable and efficient photoanodes of oxynitrides(TaON and BaTaO2N), in which highly dispersed
nanoparticles of water-oxidizing catalyst (e.g., CoOx)
loaded efficiently scavenge the photogenerated holes andeffectively suppress self-oxidative deactivation of thesurface (see Fig. 2).[6] It was also demonstrated thatco-loading of RhOx with CoOx significantly improved boththe stability and density of photocurrent on the oxynitride
photoanodes.[7] These modifications enabled us to stably
split water into H2 and O2 efficiently under visible lightirradiation at a relatively low applied bias.

Fig. 2 Water splitting under visible
lighton
CoOx-loaded
TaON
photoanode coupled with Pt electrode.

References:[1] Abe, R. J. Photochem. Photobiol. C: Photochemistry Reviews 2010, 11 179.
[2] Abe, R. Bull. Chem. Soc. Jpn. 2011, 84, 1000.
[3] a) Sayama, K.; Mukasa, K.; Abe, R.; Abe, Y.; Arakawa, H. Chem. Commun. 2001,
24169; b) Abe, R.; Sayama, K.; Sugihara, H. J. Phys. Chem. B 2005, 109, 16052.
[4] a) Abe, R.; Takata, T.; Sugihara, H.; Domen, K. Chem. Commun. 2005, 3829; b) Higashi,M.; Abe, R.; Takata, T.; Domen, K. Chem. Mater. 2009, 21, 1543; c) Maeda, K.; Higashi,M.; Lu, D.; Abe, R.; Domen, K. J. Am. Chem. Soc. 2010, 132, 5858; d) Abe, R.; Higashi,M.; Domen, K. ChemSusChem 2011, 4, 228.
[5] a) Abe, R.; Shinmei, K.; Hara, K.; Ohtani, B. Chem. Commun. 2009, 3577; b) Abe, R.;
Shinmei, K.; Koumura, N.; Hara, K.; Ohtani, B. J. Am. Chem. Soc. 2013, 135, 16872.
[6] a) Abe, R.; Higashi, M.; Domen, K. J. Am. Chem. Soc. 2010, 132, 11828; b) Maeda, K.;
Higashi, M.; Siritanaratlul, B.; Abe, R.; Domen, K. J. Am. Chem. Soc. 2011, 133, 12334;d) Higashi, M.; Domen, K.; Abe, R. J. Am. Chem. Soc. 2012, 134, 6968.
[7] Higashi, M.; Domen, K.; Abe, R. J. Am. Chem. Soc. 2013, 135, 10238.

18

Study on the mechanism of CO2 reduction with water over Ag loaded
Ga2O3 photocatalysts
Tomoko Yoshida1), Muneaki Yamamoto2), Naoto Yamamoto2) and Shinya
Yagi1)
1)
EcoTopia Science Institute,Nagoya University, Nagoya, 464-8603, Japan
2)
Graduate School of Engineering,Nagoya University, Nagoya, 464-8603, Japan
Abstract: Reduction of CO2 using a photocatalyst is an attractive reaction not only for the
mitigation of CO2 as a greenhouse gas but for the energy storage in the form of usable andvaluable products. Especially, the photocatalytic reduction of CO2 by water as the reductant isof great interest from viewpoints of an environment-friendly chemical reaction and an
artificial photosynthesis.
Recently, it has been reported that Ga2O3 shows photocatalytic activity for CO2 reduction
with water to produce CO as well as for water splitting to H2 and O2, and Ag loaded Ga2O3(Ag/Ga2O3) photocatalyst promotes the CO2 reduction [1]. However, the dynamic processesof the reaction such as the change in chemical states of adsorbates at each reaction stage are
not well understood. It is also unclear how the loaded Ag species contribute to the CO2
reduction and which chemical state and local structure of the Ag species are effective for CO
production. In order to clarify these questions, we have investigated the adsorption states of
CO2 molecules on the Ag/Ga2O3 photocatalysts during the photocatalytic reaction by in-situ
FT-IR measurements.
We have constructed the in-situ FT-IR measurement system consisting of a quartz IR cell,a heater, a gas flow apparatus and vacuum pumps. This system enables in-situ FT-IR
measurements of the sample in the various reaction conditions such as the heat pretreatment,
gas adsorption and photoirradiation.
Fig. 1(a) shows FT-IR spectrum of the
Ag/Ga2O3 photocatalyst after the introductionof CO2, and the absorption bands in the
spectrum are assigned to monodentate
bicarbonate and/or bidentate carbonate
species [2-4]. These bands are reduced after
the photoirradiation followed by evacuation,and new bands assigned to bidentate formate
species appear as shown in Fig. 1(b). CO2
molecules would react with surface hydroxyl
groups to form the monodentate bicarbonate
and/or the bidentate carbonate species, andsubsequently change to the bidentate formate

Fig. 1 FT-IR spectra of Ag/Ga2O3(a) after introduction of CO2 and (b)after photoirradiation followed by
evacuation.

19

species under the photoirradiation [2].
The bands originating from the formate species became broader by introducing water
vapors and returned to the original sharp bands after the evacuation, meaning that thechemisorbed formate species interact with water molecules. On the other hand, the bands
decreased by the photoirradiation in an atmosphere of water, and new bands derived from CO
species came out. These results suggest that the interaction of the formate species with watermolecules under photoirradiation is essential for the CO production. Accordingly, we propose
a possible mechanism for the photocatalytic reduction of CO2 with water as illustrated in Fig.
2 [5].

Fig. 2 The mechanism for the photocatalytic reduction of CO2 with over Ag/Ga2O3.

In addition, a comparison of FT-IR spectra of the Ag/Ga2O3 and bare Ga2O3 samples ismeaningful to understand how Ag species loaded on the Ga2O3 contribute to the reduction of
CO2. we found that the Ag loading enhances the elementary reactions such as the CO2
adsorption, the formate production, and the interaction between formate species and watermolecules. The Ag species would generate new active sites for the CO2 adsorption and thesubsequent conversion into formate species.
References:[1] Z. Like, E. Tanabe, K. Morikawa, T. Kazino, T. Sekihuzi, S. Matsumoto, Y. Hirata and H.
Yoshida, 108th Symposium of Catal. Soc., Kitami, Japan, 2011, pp 1D15.
[2] H. Tsuneoka, K. Teramura, T. Shishido and T. Tanaka, J. Phys. Chem. C, 2010, 114,
8892.
[3] G. Busca, V. Lorenzelli, Mater. Chem., 1982, 7, 89.
[4] J. I. Di Cosimo, V. K. Diez, M. Xu, E. Iglesia, and C.R. Apesteguia, J. Catal., 1998, 178,
499.
[5] M. Yamamoto, T. Yoshida, N. Yamamoto, H. Yoshida and S. Yagi, e-J. Surf. Sci.
Nanotech., 2014, 12, 299.

20

OCARINA; Past, Now and Future    
Hideki Hashimoto (Project Leader)
The OCU Advanced Research Institute for Natural Scienceand Technology (OCARINA),
Osaka City Univeristy, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan
Abstract: The Osaka City Univeristy Advanced Research Institute for Natural Science andTechnology (OCARINA) was offcially founded in April 2007 owing to the devoted effort bylate Prof. Shohachi Kakuno who was the vice-president of Osaka City University at that time.
The OCARINA started as a virtual institue at the beginning. However, the offcial regulations
and by-law of OCARINA were developped in March 2010, and the research initiative wasplaced in 2-Gokan building at general education campus in October 2010. OCARINA is now
well recognized as the distingusihed research institute that respresents the presence of Osaka
City Univeristy. The establishement of OCARINA is closely related to the construction of
new scientific research buildings as well as the Research Center for Artificial Photosynthesis(ReCAP), both of which are greatly supported by Osaka City. In this talk, as the first project
leader of OCARINA during the fiscal years between 2007 – 2014, I will summrize the historyof why and how OCARINA has been expanded. I want to postulate the future perspective of
OCARINA for the purpose to hand my position to the New Project Leader.

21

Thursday, 5th March

22

23

Materials

Photomechanical Actuators for Next Generation Intelligent
Seiya Kobatake
Graduate School of Engineering, Osaka City University

Photochromic compounds undergo a photochemically reversible transformation reaction
between two isomers. Such molecules in organic crystals, which are regularly oriented andfixed in the crystal lattice, may be potentially useful for optoelectronic devices. To apply
photochromic diarylethene crystals to photonics, electronics, mechanics, and medical fields,
the materials are required to change large physical property by photoirradiation. We foundthat small size microcrystals exhibit photochromism in high conversion and show large
physical property changes such as crystal deformation [1]. In this paper, we have focused on
photochromism of the diarylethene crystals and their photomechanical phenomena, such as
crystal bending and crystal twisting.
Diarylethene derivaties are desinged and synthesized for study on the photoinduced
crystal shape changes. Upon irradiation with UV light to the diarylethene crystals, themolecules in the crystals underwent a photocyclization reaction that transforms the initialopen-ring isomers into the photogenerated closed-ring isomers in the crystalline phase. The
rod-like crystal of diarylethenes showed the crystal bending away from the incident UV light[2] or toward the incident UV light [3]. The bending speed depended on the crystal thickness
and the curvature change against the crystal thickness was well fitted to Timoshenko’s
bimetal model. The bent crystal returned to the initial straight crystal by irradiation withvisible light. Furthermore, we have found specific crystal ribbon deformation. This crystal
reversibly changed from the straight thin crystal to the spiral twisting crystal [4]. Suchdeformation depends on the crystal packing. These materials can be potentially applied fornovel organic solid devices, and it is possible not only to become a new organic device but
also to open a new possibility as the crystal materials.
UV‘‘ON’’‘‘OFF’’

1 mm

Figure.

1 mm

Examples of photomechanical actuators we published.

References[1] Kobatake, S.; Takami, S.; Muto, H.; Ishikawa, T.; Irie, M. Nature 2007, 446, 778.
[2] Kitagawa, D.; Kobatake, S. J. Phys. Chem. C 2013, 117, 20887.
[3] Kitagawa, D.; Kobatake, S. Photochem. Photobiol. Sci. 2014, 13, 764.
[4] Kitagawa, D.; Nishi, H.; Kobatake, S. Angew. Chem. Int. Ed. 2013, 52, 9320.

24

Application of low-temperature bondings for novel device
process technologies targeting greener societies
Naoteru Shigekawa and Jianbo Liang
Department of Applied Physics and Electronics,
Graduate School of Engineering, Osaka City University
Low-temperature bondings are promising for fabricating novel functional devices such as
hybrid multi-junction solar cells since these technologies enable us to make heterostructuresof semiconductor materials with different crystal structures, lattice constants, and thermal
expansion coefficients [1]. In this presentation, we report on characteristics of
low-temperature-bonding based InGaP/GaAs/Si hybrid triple-junction solar cells, which areassumed to bring about high conversion efficiencies with low costs in photovoltaics [2]. We
also discuss the possibilities for realizing novel devices that could overcome the trade-offs inpower device characteristics between their power capabilities and switching speeds.
We bonded InGaP/GaAs double-junction solar cell structures grown on GaAs substrates to
Si-based bottom cells after activating their surfaces without heating. Then we madetriple-junction cells by selectively etching GaAs substrates and subsequently employing theconventional semiconductor device process sequences. Fabricated cells revealed theenhancement in cell characteristics in comparison with those of the respective sub cells. A
conversion efficiency of 24.4% was achieved [2]. We also fabricated Si/SiC pn junctions
using the low-temperature bondings and observed improvements in their diode characteristics
due to post-bonding annealing [3]. These results indicate that the low-temperature bonding
methods should play an essential role in constructing novel device process technologies forrealizing greener societies.
[1] O. Moutanabbir and U. Gösele, Annu. Rev. Mater. Res. 40, 469 (2010).
[2] N. Shigekawa, L. Chai, M. Morimoto, J. Liang, R. Onitsuka, T. Agui, H. Juso, and T.
Takamoto, Proc. 40th IEEE Photovoltaic Specialists Conference, pp. 534-537 (2014).
[3] J. Liang, S. Nishida, M. Arai, and N. Shigekawa, Appl. Phys. Lett. 104, 161604 (2014).

25

Smart Energy Networks for Near Future Cities    
photoMasaki Nakao
Project Professor, 3-9-11,Sugimoto,Sumiyoshi-ku,Osaka City, Japan

Abstract:
To secure a stable energy supply, low carbon usage for existing urban areas, we aredoing research to build highly efficient distributed energy resource systems that enable newand existing buildings to use renewable energy. Project target is to realize a flexible
interchange of heat for smart communities, to increase renewable energy, and to make it easy
to use, as follows.
�Thermal routing technology that minimize primary energy consumption and trade thermalenergy between suppliers and consumers in heating and cooling of buildings
�Increasing technology of bio-gas production in waste water treatment plant using heat fromsolar, digested sludge, and treated water�Aquifer thermal energy storage technologies having both daily and seasonal storage of cold
and/warm groundwater in an aquifer�Apply hydrogen energy in the near future to the distributed energy resource systems
�Apply developed sewer heat recovery technology to them at manhole along sewer pipe line
�Integration of energy source, for example, hydrogen, bio-gas, groundwater, and sewer heatetc. for urban smart energy supply and demand systems
The goal of this project is to develop the basic technologies and demonstrate aworking system. Results for this year are as follows.
We have completed the actual measurement of air conditioning system in the target
buildings. From cooling load analysis during the summer season, it is shown that there arezones of partial cooling load, and since the chilled water return temperature of the partial
cooling load zone is not high, the return chilled water could be supplied to the other partial
cooling load zone. That is called chilled water cascading. For example, even in the peakcooling load in the summer, 57% of cooling capacity of chilled water is cascadable to theother buildings, according to the measurement data of the ATC building. CO2 reduction was
calculated to 9200t-CO2 / year.
It is necessary to control the chilled water supply route for cascading. We havedeveloped a thermal routing technology to flexibly change the combination of supply side anddemand side of chilled water supply route in accordance with the operating condition of the
air-conditioning system, and construct a practical demonstration system. In addition, we

26

started to develop thin-walled pipes with inner insulation to reduce the heat loss and heat
capacity. We have created an algorithm for the optimization of the heat source as subset of
EMS. By controlling the combination of the energy demand side and the supply side, it isshown that energy consumption of the heat source can be reduced up to 75%.
In the development of increasing technology of bio-gas production in waste water
treatment plant, we built a prototype "smart valve" for the coordinated control of multiple
solar-powered water heaters by independently controlling valves using signals from themulti-sensor. Heat collection temperature of solar thermal panel can be regulated fairly
constant by switching the connection of the solar panel in parallel and series with these smart
valves. Developed heat pump can achieve high-efficiency with high temperature output, andexamined control logic for multiple heat pumps. We studied heat exchanger for heat recovery
from sludge, and developed film flow heat exchanger and evaluated their performance using
simulated sludge.
The City of Osaka is promoting the ‘Sakishima District Smart Community
Demonstration Project” as part of its energy policy within the Kansai Innovation International
Strategic Comprehensive Special Zone. One of the aims is to stimulate the area by involving
new players in this energy project. We can contribute to this Smart Community
Demonstration Project through our R & D.

27

Bridging protein in gliding machinery of Mycoplasma mobile    
Tasuku HAMAGUCHI1, Yuhei O. TAHARA1 and Makoto MIYATA1,2
1)Graduate School of Science, Osaka City University.
2) OCARINA, Osaka City University
Mycoplasmas cause various deseases in plants and amimals including human pneumonia andurethritis. Some mycoplasmas form protrution at a cell pole and glide on solid surfaces
including their host cells. This gliding is diverged and generated by a unique machinerywith internal and surface structures. Mycoplasma mobile, a fish pathogen, is the fastest
gliding species with a speed of 2.0~4.5 µm/s. Jellyfish structure, an internal structure of M.
mobile, is composed of dozens of genes including F1-ATPase paralogs and functionary
unknown proteins. F1-ATPase paralogs hydrolyze ATP and generate the force for gliding[1]. Three large proteins, Gli349 (Leg), Gli521 (Crank) and Gli123 (Mount), are involved in
the surface of the gliding machinery [2]. However, the association between the internal andthe surface structures across the cell membrane is still unknown. In this study, we found anovel component protein possibly bridging between internal structure and surface structure ofgliding machinery.
Gli521 was isolated from M. mobile cells associating with MMOB1650, a component of theinternal structure with molar ratio of 2.5 : 1. The amino acid sequence of MMOB1650
composed of 1158 amino acids were featured with one and two transmembrane segments at
N- and C-terminus regions, respectively.
Electrophoretic analyses suggested that
MMOB1650 was truncated by protein processing.
Crosslinking by usingbis(sulfosuccinimidyl)suberate and mass spectrometry suggested that N- and C-terminus
regions of MMOB1650 interact with the “Hook”, C-terminus region of Gli521. Electron
microscopy suggested that MMOB1650 and Gli521 complex appeared a structure similar totrimeric structure of Gli521, attached with an extra density at the center. Based on theseresults, MMOB1650 was suggested to bridge the internal structure to Gli521, crank on thesurface of gliding machinery.
[1] Tulum, I., Yabe, M., Uenoyama, A. and
Miyata, M. Journal of Bacteriology, 2014, 10,01418.
[2] Miyata, M. Annual Review ofMicrobiology, 2010, 64, 519.

28

Charcterization and Engineering ofanimal opsins as optogenetic tools    
Akihisa Terakita and Mitsumasa Koyanagi
Department of Biology and Geoscience, Graduate School of Science, Osaka City
University, Osaka 448-8585, Japan
Abstract: Animals capture light information through visual pigment rhodopsin and related
light-sensor proteins, opsins and utilize them for visual and non-visual functions including
regulation of circadian rhythms. Opsin is one of G protein-coupled receptors (GPCRs),
which constitute a large protein family of receptors that sense molecules outside the cell andactivate inside signal transduction pathways and, ultimately, generate cellular responses. In
photoreceptor cells, opsin binds to a chromophore retinal to form pigment and functions as alight-sensitive GPCR.
Thausands of opsins have been identified from a wide variety of animas. We havecharacterized various opsins, including novel ones, spectroscopically, biochemically andmolecular physiologically [1-3], and also investigated their contribution to biological function[4]. Interestingly, we found that some non-visual opsins have unique molecular propertes
suitable for their function in the extraocular tissues. A “brain opsin” binds to a chromophore
retinal present ubiquitously even in extraocullar tissues and also can respond to light
repeatedly, showing the reusable property. We also found a novel opsin that light-dependently
drives a signal transduction cascade involving elevation of inctracellular cAMP level,
important second messenger in various cells. The molecular characteristics of these opsinssuggest their potentials for optogenetics, which is the combination of genetics and optics tocontrol well-defined events within specific cells of living tissues and is now a strong
application to control cell activities and animal behaviors by light.
Here, we introduce the aim of our project; that is, developing new optogenetic tools which
control GPCR-cAMP-signalling in the various cells, by using opsin having properties suitablefor functioning in extraocullar tissues. We try to engineer chimera and/or mutant opsins that
respond to light repeatedly and also sense different color of light to regulate cAMP cascade,and then show their availability in animal models. Further, we are also planning toinvestigate their 3D structures as a international collaboration to provide structural basis forthese new optogenetic tools
References:[1] Tsukamoto et al., J. Biol. Chem 2009, 284, 20676[2] Koyanagi et al., Proc. Natl. Acad. Sci. U.S.A 2008, 105, 15576.
[3] Koyanagi et al., Proc. Natl. Acad. Sci. U.S.A 2013, 110, 4998
[4] Nagata et al., Science 2012, 355, 469

29

Advanced ESR Spectroscopy as Molecular Spin Technology
for Materials Innovations
Kazunobu Sato
Graduate School of Science, Osaka City University
Abstract: Environmental and energy resource issues have been emerging in the last decades.
It is expected that innovations based on modern science and technology can afford tocontribute to solve such global materials issues. We have implemented new-paradigm based
Electron Spin Resonance (ESR) spectroscopy to molecular synthetic chemistry to realize
advanced spin technology, manipulating microscopic quantum nature of molecular open-shell
systems [1-4]. In this project, we focus on two topics relevant to organic spin mediatedsecondary batteries and molecular spin quantum computers, aiming to develop innovative
quantum spin technology based on ESR and to apply to molecular spin systems for thesustainable growth in the future.
Secondary batteries have been widely applied to various mobile electronics since Lithium
Ion secondary Batteries (LIBs) appeared. Recently, the risk of the conventional LIBs for theuse of lithium and rare metals has been recognized. A materials challenge for rare-metal freesecondary batteries is also one of the current issues from the viewpoint of mineral resourcesand our economic environments. Organic secondary batteries have attracted considerable
interest as a tailored-made materials system for overcoming the risk of safety, resource costs
and environmental issues inherent in the conventional lithium-ion batteries. We havedeveloped organic rechargeable batteries with both high current capacity and cycle
performance [1]. In addition to electrochemical approaches, the importance of microscopic
analyses for the batteries has been recognized. Real-time/in-situ and spectroscopic
measurements applicable to the batteries have just started in order to understand battery
reactions inside the batteries during charge/discharge processes. We have carried out theimplementation of organic secondary batteries as well as the understanding of the battery
action in microscopic terms by using advanced ESR methods such as in-situ measurements
and electron spin imaging of the batteries.
Realization of practical quantum computing and quantum information processing(QC/QIP) has been challenging topics in advanced scientific fields, particularly quantum
technology. Quantum computing processes which are reversible state transformations are
considered as extremely favorable ones without energy/information loss. For the last morethan ten years, we have been studying the implementation of molecular-spin quantum
computers based on molecular open-shell systems by applying pulsed microwave/RF spintechnology [3,4]. Recently, we have achieved the development of NMR-paradigm ESR
spectroscopy, implementing molecular spin quantum operations.
References:[1] Y. Morita, S. Nishida, K. Sato, T. Takui et al., Nature Materials 2011, 10, 947.
[2] Y. Morita, S. Suzuki, K. Sato, and T. Takui, Nature Chemistry 2011, 3, 197.
[3] K. Sato, S. Nakazawa, Y. Morita, T. Takui et al., J. Mater. Chem. 2009, 19, 3739.
[4] S. Nakazawa, K. Sato, Y. Morita, T. Takui et al., Angew. Chem. Int. Ed. 2012, 51, 9860.

30

π-Figuration of Open-shell Molecular Systems
for Material Innovation    
Shuichi Suzuki,1,2 Masatoshi Kozaki,1 Keiji Okada1,2
1
Department of Chemistry, Graduate School of Science, Osaka City University, 2OCARINA
Molecule-based materials underlie promising next-generation electronic devices, nanoscalemachines, and quantum information processing systems because of their wide diversity inmolecular designs. In particular, open-shell molecular systems have recently attracted
considerable attention because of their unique functionalities based on electron spins andredox properties. The most widely used spin sources in molecular materials are metal ionsbecause of their stability and large spin multiplicities. However, the unpaier electrons in themetal-ligand systems are largely localized on the metal ions, and the spin-spin interactions
between the metal ions are expected to be very small. Thus, it is essential for materials
innovation using molecule-based materials to design new delocalized-spin molecular systems.
We have recently studied on new synthetic routes of open-shell molecules based on

π-conjugated systems [1-4]. The purpose of this project is designs and syntheses of new
π-conjugated open-shell molecular systems with unique functionalities by developing our
synthetic methods.

References:[1] Suzuki, S.; Okada, K. et al. J. Am. Chem. Soc. 2010, 132, 15908.
[2] Suzuki, S.; Okada, K. et al. J. Am. Chem. Soc. 2012, 134, 17866.
[3] Suzuki, S.; Okada, K. et al. Eur. J. Inorg. Chem. 2014, 4740.
[4] Suzuki, S.; Okada, K. et al. Chem Lett. 2014, 43, 678.

31

Biofuel production in a CO2-absorbing microalga
Euglena gracilis
Masami Nakazawa1,2, Mitsuhiro Ueda1,
Hiroshi Inui1, Yoshihisa Nakano1, Kazutaka Miyatake1
1. Osaka Prefecture University, 2. PRESTO, JST.
Abstract: Euglena is a protist (unicellular eukaryote) with both "plant-like" and "animal-like"characteristics in its metabolism. Euglena gracilis, a species of Euglena, grows under both
autotrophic and heterotrophic conditions. We previously found E. gracilis have high CO2
tolerance. Growth of E. gracilis was enhanced under 5-40% concentration of CO2 comparedto growth in the air (0.04% CO2). These characteristics are important for CO2 sequestration
systems, especially in the direct utilization of power plant flue gas. Excess amount ofcarbohydrates from photosynthesis and extracellular carbon sources are converted toparamylon, a kind of insoluble β-1,3-glucan, and stored in cytosol. Paramylon syntheses
mainly occur in the aerobic conditions under both autotrophic and heterotrophic cultivations.
E. gracilis adapt to a broad range of oxygen concentrations and can survive evenvery low concentrations of oxygen. When aerobically grown E. gracilis expose to hypoxicor anaerobic conditions, they promptly synthesize wax esters with the concomitant decreaseof paramylon in the cell [1,2]. Surprisingly, the anaerobic wax ester formation isaccompanied by a net synthesis of ATP from glycolysis. We named this phenomenon wax
ester fermentation. A malonyl-CoA independent fatty acid synthesis occurs in mitochondriaof E. gracilis and the system has ability to synthesize fatty acids directly from acetyl-CoA as
both primer and C2 donor. The wax esters produced by E. gracilis under anaerobic
conditions are composed of saturated fatty acids and alcohols, the main ones being myristic
acid and myristic alcohol, respectively. These are expected as materials of algae-based jet
fuel. The yields of the wax esters are different in various medium compositions, ranging
around 30-40% (w/w dry cell). To achieve higher wax ester production for industrial use,
we investigated detail metabolic pathway to determine possible rate-limiting steps in the wax
ester production. In this talk, we are going to focus 1) hypoxic production of acetyl-CoA by
the unique oxygen-sensitive pyruvate:NADP+ oxidoreductase (PNO) in mitochondria and 2)
hypoxic fatty acid production in mitochondria.
References:[1] Inui, H.; Miyatake, K.; Nakano, Y.; Kitaoka, S. Eur. J. Biochem. 1984, 142, 121–126.
[2] Inui, H.; Miyatake, K.; Nakano, Y.; Kitaoka, S. Arch. Biochem. Biophys. 1989, 274,
434–442.

32

CytosolParamylon(ß-1,3 glucan)

Wax ester

glucose unit
glycolysis

Fatty alcohol

PyruvateER

Pyruvate
PNO

Found only in Euglena

Acetyl-CoA

Acyl-CoA        Mitochondrion
FigureDeduced pathway of hypoxic wax ester production from storage polysaccharide, paramylon

33

Current achievements of “e-Lab”    
Shigeyuki Minami and Hiroshi Ikeda
Advanced Research Insutitute for Science and Technology, OCU
Abstract: Recent achievements are reported here on the project of “e-Lab”, which aims forthe following four research objects: (1) Electric energy, (2) Environment, (3) Elderly care, (4)Education for electrical engineers. Firstly, it was found that a remarkable CO2 reduction could
be accomplished for fishing if our developed plug-in hybrid electric fishing boat is used. This
research is conducted under the research program of “Empirical study of the independence
distribution energy system technology based on DC technology in a detached island and afishing village” financed by the Ministry of Environment of Japan subsidy 2014-2016.
Secondly, the current status of our newly developed hydrogen propulsion system for marine
application is introduced. Our research results for elderly or physically handicapped persons
are introduced. For an educational contribution to electrical engineers, our aurora display
device is also developed.
(1) On the development of Plug-in Hybrid Boat and S2G Research
We have demonstrated a newly developed S2G two-way electricity transportation system(from ship to grid and from grid to ship) to prove the effective use of renewable energy forship propulsion and a successful reduction of CO2 emission. This research is conducted under
the research program of “Empirical study of the independence distribution energy systemtechnology based on DC technology in a detached island and a fishing village” financed by
the Ministry of Environment of Japan subsidy 2014-2016.
To conduct the research, a plug-in hybrid electric fishing boat (50 kW electric motor, 140 kW
diesel engine with a 10 kW generator and a 20 kWh Li-ion battery system) was prepareddeveloped under collaboration research between OCU and the Kansai Electric Power Co., Inc.
One electric vehicle (total battery of 20 kWh) was also used to transport the electric energy
obtained by a solar cell power line (the grid) with the Li-ion 46 kWh battery system to theboat as the normal G2S operation. The PHEB can also generate electricity so that the electricenergy can be supplied from the boat to the grid via the electric vehicle when natural disasters
happen.
It is shown that the transportation efficiency of the S2G system from the ship to the grid andvice versa was kept at 90 %. It is also quantitatively shown that the PHEB provides an
outstanding CO2 reduction of more than 80 % compared with the simple diesel engine
operation when a professional fixed position fishing pattern at the research island was
conducted. The remarkable reduction of oil consumption is due to the high efficiency of theelectric motor and the good performance of the idling stop function of the electric motor
propulsion during fixed fishing operation [1, 2]

34

(2) Newly Developed Hydrogen Propultion System
No new fundamental system for ship propulsion has been discovered for years. Our researchaims to develop a new ship propulsion system using hydrogen fuel. This propulsion device
uses hydrogen ignited in water for propulsion. It is proved that a model boat with this system
worked successfully by a gaseous high temperature reaction without making noise in thewater. This system can be used for future new boat propulsion.
(3) e-Lab Research
Osaka City University has many talented researchers on energy research, environmental
research, elderly people’s care and engineering education. The e-Lab research was foundedbased on the above mentioned four research objects matching the aim of OCARINA. Thisassimilation provides many fruitful results on the four research objects. The details and thecurrent status of e-Lab are introduced.
Our research program for elderly or physically handicapped persons is to povide contributions
to society. Some of the following current achievements related to the welfare are introduced.
(a) Designing a nap room in the vehicle for professional bus drivers's fatigue reduction andquiet sleep. (b) Development of an operation controller for vehicle equipment which enables
easy driving by a driver who is physically handicapped.
As one of the contributions to education for engineers, we have designed aurora simulation
devices at 13 scientific facilities including the Shanghai EXPO 2010.
References:[1] Minami, S.; et. al., On the performance of a newly developed plug-in hybrid boat (PHEB),
Journal of Asian Electric Vehicles 2013, Vol. 11, No. 2, 1653-1657.
[2] Minami, S, et al.; Performance of a newly developed plug-in hybrid boat, Proceedings of
EVS27, Barcelona, Spain, 2013.
[3] Ikeda, H., et al., Development of body support equipment for effective driving for
persons with a limb/trunk dysfunction, Studies in Science and Technology 2013, Vol. 2,
No. 2, 147-150.

35

36

Poster Presentations37

P1
Effect of Clostridium butyricum MIYAIRI588
upon Lifespan and Stress Resistancein Caenorhabditis elegans    
Maiko Kato1, Tomomi Komura1, Eriko Kage-Nakadai1,2, Yoshikazu Nishikawa1
1
Graduate School of Human Life Science, Osaka City University, Osaka 588-8585, Japan
2
The OCU Advanced Research Institute for Natural Science and Technology
Abstract: [Purpose] We examined whether diet could delay senescence of host defense
mechanisms and extend the healthspan of Caenorhabditis elegans, which is a usefulnematode for aging research. Clostridium butyricum MIYAIRI588 (CBM588), which isGram-positive, obligately anaerobic, and spore-forming bacterium, has been reported toinhibit the development of pathogenic bacteria. After we reported that oral supplementation
with lactic acid bacteria extends lifespan of C. elegans [1], many studies on probiotics havebeen performed by using C. elegans models. However, there are no studies that showed thelongevity effect of C. butyricum to our knowledge. Hence, in this study, we fed the worm on
CBM588 and investigated how the clostridia influenced the lifespan and resistance to various
stresses.
[Methods] Hermaphrodite C. elegans Bristol N2 strain synchronized by an egg preparation
were fed standard food, Escherichia coli OP50 strain, on modified nematode growth medium
until they reached 3-day old. Then, animals were fed OP50 alone, CBM588 alone or a
mixture of OP50 and CBM588, respectively, and the lifespan, body size, and brood size were
measured. Resistance against stresses, such as heat, hydrogen peroxide, copper ion, ultraviolet,
paraquat, and Salmonella Enteritidis or Staphylococcus aureus infection, were compared
between the worms fed on OP50 and those on CBM588.
[Results and Discussion] The worms fed on CBM588 alone exhibited significantly extended
lifespan while the body size and brood size were smaller than those of the worms fed E. coli
OP50; CBM588 would prolong the lifespan by dietary restriction. However, the mixture of
CBM588 and OP50 did not affect the body size and increased the brood size. These results
suggest that the longevity effect of CBM588 is not simply due to dietary restriction. In
addition, resistance to salmonella or staphylococci infection, copper ion, heat, and UV wasenhanced by CBM588. The clostridia presumably prevent senescence of the host defense in C.
elegans and extend the lifespan.
References:[1] T. Ikeda, C. Yasui, K. Hoshino, K. Arikawa, Y. Nishikawa. Appl Environ Microbiol 2007
Oct, 73(20):6404-9.

38

P2
Synthesis and Bioactivity of Kaitocephalin Analog Bearingan Alkyl Side Chain on the Aromatic Ring
Yuya Yoshida, Yoko Yasuno, and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Kaitocephalin (1), isolated from the filamentous fungus Eupenicillium Shearii
PF1191 by Seto and Shin-ya et al., is known to be a potent antagonist of ionotropic glutamate
receptors (iGluRs).[1] iGluRs play a role in the excitatory neurotransmission, the constructionof memory and learning, and occasionally cause ischemic neuron death. Therefore,
antagonists of iGluRs are regarded as a potent drug lead to develop therapeutic agents for thetreatment of neurological diseases such as Alzheimer’s and Parkinson’s diseases. As an aim to
discover ligands with high affinity and selectivity, we devised a bioassay system using afluorescent kaitocephalin analog. In this presentation, we would like to report our synthetic
efforts toward development of fluorescence labeled analog of 1 in which the chloro group on
the aromatic ring is substituted with other functional groups (e.g. 2-5) based on previous
structure-activity relationship study[2], and X-ray crystal structure of 1 bound to the ligand
binding domain of AMPA receptor subtype of iGluRs.[3]1: R = Cl (Kaitocephalin)
HOR

2: R =
H
N

Cl
O

H
CO 2H

CO 2H
NH 2

N
H
HO

CO2H

CO 2H

3: R =
4: R =
5: R =

On(now running)[1] Shin-ya, K.; Kim, J.-S.; Furihata, K.; Hayakawa, Y.; Seto, H. Tetrahedron Lett. 1997, 38,7079.
[2] 51th Symposium on the Chemistry of Natural Products 2009, 401.
[3] Ahmed, A. H.; Hamada, M.; Shinada, T.; Ohfune, Y.; Weerasinghe, L.; Garner, P. P.;
Oswald, R. E. J. Biol. Chem. 2012, 287, 41007.

39

P3
Synthetic Study of Tetrodotoxin
Atsushi Manabe and Tetsuro Shinada
Graduate School of Sci., Osaka City Univ.
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Tetrodotoxin (1), originally isolated from the puffer fish, is well-known marine toxin,
and attracts much attention from scientists because of its unique chemical structure and potent
biological activities. Biological mechanism of 1 was reported to be a specific inhibition ofvoltage-dependent Na+ channels. Since this finding, 1 has widely used as an importantbiochemical tool to investigate neurophysiological studies. The first total synthesis of rac-1
was reported by Kishi, Goto, et al. in 1972. [1] After that, the total synthesis of optically
active 1 have been reported by three groups, Isobe and Nishikawa, Du bois, and Sato groups,
independently. [2]
HO
H
H2N
N
H N
O
H

O
H O
O
O
OH
OH

Tetrodotoxin (1)In this presentation, we would like to describe our recent efforts toward the total
synthesis of 1 via Diels-Alder (DA) reaction as a key step. Oxabicyclo[2,2,1]heptane
derivative 3 was synthesized by DA reaction of nitroolefin 2 and 3-aminofuran in one step.
The quaternary amino carbon-center was constructed by a palladium-catalyzed allylation
reaction to give 4. Heteroatom manipulation of 4 followed by a ring opening reaction of thebridge head moiety provided 5 as a key intermediate.
O
O 2N

O 2N
NMeBocCO2 Et2

CO2Et
O

1N HCl
3

O

O2N

Pd
OAc

OTBS

OTBS NsHN
O
4 OH

OTBS
5

1

O

[1] (a) Kishi, Y.; Aratani, M.; Fukuyama, T.; Nakatsubo, F.; Goto, T.; Inoue, S.; Tanio, H.;
Sugiura, S.; Kakoi, H. J. Am. Chem. Soc. 1972, 94, 9217. (b) Kishi, Y.; Fukuyama, T.;
Aratani, M.; Nakatsubo, F.; Goto, T.; Inoue, S.; Tanio, H.; Sugiura, S.; Kakoi, H. J. Am.
Chem. Soc. 1972, 94, 9219.
[2] (a) Nishikawa, T.; Urabe, D.; Isobe, M. Angew. Chem., Int. Ed. 2004, 43, 4782. (b)Hinman, A.; Du Bois, J. J. Am. Chem. Soc. 2003, 125, 11510. (c) Akai, S.; Seki, H.; Sugita,
N.; Kogure, T.; Nishizawa, N.; Suzuki, K.; Nakamura, Y.; Kajihara, Y.; Yoshimura, J.; Sato,K. Bull. Chem. Soc. Jpn. 2010, 83, 279.

40

P4
Total synthesis of Cirratiomycin A    
Akito Nishimura, Yoko Yasuno and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Cirratiomycin A (1) exhibiting a characteristic antibiotic spectrum was independently
isolated from Streptomyces cirratus 248-Sq2 by Seto et al.[1a], and Streptomyces
xanthocidicus MG125-CF1 by Umezawa et al..[1b] One of the key to the total synthesis is thestereoselective synthesis of E-dehydroisoleucine (E-ΔIle).
We would like to report the first total synthesis of 1 by the stereoselective synthesisof E-ΔIle-containing tripeptide fragment 4[2] and its coupling with other fragments 2 and 3.

O
H 2N

N
H
OH OH

O

H
N
O
H 2N

H
N

O

O

N
N

N
H

H
N

CO2H

O

OH

H
Cirratiomycin A (1)

O
BocHN
O

O
N
H

O
2

CO2H

FmocHN
BocHN

CO2 H
N
N

O
BocHN

N
Boc

H
3

H
N
O

CO 2MeOH4[1] (a) Shiroza, T.; Ebisawa, N.; Kojima, A.; Furihata, K.; Shimazu, A.; Endo, T.; Seto, H.;
Otake, N. Agric. Biol. Chem. 1982, 46, 1885−1890. (b) Shimada, N.; Morimoto, K.;
Naganawa, H.; Takita, T.; Hamada, M.; Maeda, K.; Takeuchi, T.; Umezawa, H. J. Antibiot.
1981, 34, 1613−1614.
[2] 94th Annual Meeting of the Chemical Society of Japan, 3H6-47 (2014).

41

P5
Synthetic Study of Natural Products with a Cyclic Tripeptide Core    
Yoko Yasuno, Shunsuke Yamaguchi, Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Phomopsin A (1)[1] and ustiloxin D (2)[2] are known to be potent antimitotic agentsthat bind to tubulin and inhibit microtubule assembly. These natural products are featured bytheir 13-membered cyclic tripeptide core which is speculated as the pharmacophore for the
tubulin binding.[3] Herein, we would like to report our recent efforts toward stereoselective
synthesis of the tripeptide core of 1 and 2. The application to the synthesis of ustiloxin analog
3 bearing a glutamic acid side chain will be described.
Me

MeHN

HN
O O Et
Me
O

HO

Cl

OH

Me

O

NH
N

H
N
Me

O
Phomopsin A (1)

O

Et

N
H

Me

MeHN

HN

HO

CO2H

O O Et
Me
O

CO2H

OH

H
N

CO2H

O

Ustiloxin D (2)

Me
MeHN

HN

HO

O O Et
Me
O

OH

NH

Me
NH

H
N

CO2H

3

CO2H

O

[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, 269–277.
[2] (a) Koiso, Y.; Natori, M.; Iwasaki, S.; Sato, S.; Sonoda, R.; Fujita, Y.; Yaegashi, H.; Sato,Z. Tetrahedron Lett. 1992, 33, 4157–4160. (b) Koiso, Y.; Li, Y.; Iwasaki, S.; Hanaoka, K.;
Kobayashi, T.; Sonoda, R.; Fujita, Y.; Yaegashi, H.; Sato, Z. J. Antibiot. 1994, 47, 765–773.
[3] Morisaki, N.; Mitsui, Y.; Yamashita, Y.; Koiso, Y.; Shirai, R.; Hashimoto, Y.; Iwasaki, S.
J. Antibiot. 1998, 51, 423–427.

42P6
Synthetic Study of Kainoids by an Intramolecular Cyclization    
Azusa Yamada and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Kainic acid (1) and its derivatives such as dihydrokainic acid (2) are known to beagonists of ionotropic glutamate receptors. These are unusual cyclic amino acids possessing apyrrolidine ring with three contiguous substituents. Kainoids have attracted much attention assynthetic targets in view of their pharmacological activities as well as unique structuralfeatures. Previously, we have reported a novel synthetic approach to construct the pyrrolidine
ring by an intramolecular radical cyclization for the facile access to the kainoid framework.[1]In this presentation, we will report the synthesis of 2.
CO2H

CO2H
CO2H

N
H

N
H

CO2H

Previous Work
R1
R2

R1

OBn

CO2Me
N
Boc

CO2Me
N
Boc

Dihydrokainic acid (2)Kainic acid (1)I

intramolecular2
OBn radical cyclization R

Diester 4 was synthesized from 3. Treatment of 4 with KHMDS induced
epimerization at C2 to give the protected dihydrokainate 5. Removal of the protecting groupsof 5 afforded the target compound 2.
OBn

CO2Me

KHMDS

CO2Me

CO2Me
N
Boc

CO2Me
N
Cbz

THF, 0 °C

CO2Me
N
Cbz

3

4

2

2

5

[1] The 2013 OCARINA Annual International Meeting, P55 (2014).

43

P7
The relationship between SIR-2.1 and the longevity effect of sesaminin Caenorhabditis elegans    
Yumiko NAKATANI 1, Yukie YAGUCHI 1, Noriko KASHIMA 1, Tomomi KOMURA 1,
Eriko KAGE-NAKADAI 1,2, sigeru SAEKI 1,Keiji TERAO 3, Yoshikazu NISHIKAWA 1
1
Graduate School of Human Life Science and, 2 The OCU Advanced Research Institute for
Natural Science and Technology, Osaka City University, Osaka 558-8585, Japan;
3
CycloChem Co., Ltd, Kobe 650-0047, Japan
Abstract[Introduction]
Sesamin is one of the lignans present most abundantly in sesame seed and oil. We previously
reported that oral administration of sesamin extended the lifespan and conferred resistance tothe stresses such as hydrogen peroxide, paraquat, or opportunistic infection with legionella byusing C.elegans model [1].C.elegans mutant analyses suggested that the effects of sesaminwere mediated by p38 MAPK and insulin/IGF-1 pathways. In addition, sesamin triggered thenuclear localization of transcription factor DAF-16 that is a homolog of mammalian FOXO.
C.elegans SIR-2.1 is a member of the sirtuin family related to Saccharomyces cerevisiae
Sir2p that has been implicated in the senescence. The mammalian homolog SIRT1 plays
important roles in multiple cellular processes including transcriptional repression and stressresponse. SIR-2.1 has been shown to combine with DAF-16 in the nucleus and triggeringtranscriptional activation of DAF-16-target genes upon stress.
In this study, we investigated the relationship between SIR-2.1 and the longevity effect ofsesamin.
[Materials and Methods]
Nematodes were fed the bacterium Escherichia coli strain OP50 in the mutant studies, or thebacterium E.coli strain HT115 containing RNAi constructs in the RNAi studies. Synchronizedworms were fed the bacteria with or without sesamin/ɤ-cyclodextrin inclusion compounds
from three-day-old adults, and the mean life span was calculated. Transcriptional changes
elicited 24 h after the intake of sesamin were measured by microarray analysis andquantitative RT-PCR (qRT-PCR).
[Results and Discussion]
Loss-of-function mutation in sir-2.1 diminished longevity effects of sesamin. We confirmedthat RNAi knockdown of sir-2.1 similarly canceled the effects. These data supported the ideathat sesamin may activate the SIR-2.1 pathway. Because the longevity by SIR-2.1
overexpression requires autophagy, we investigated whether autophagy-related genes were44

P7
involved in the effect of sesamin. As a result, the RNAi knockdown of bec-1, which isessential for the autophagosome formation, diminished the longevity effect of sesamin.
Taken together, our findings suggest that sesamin extends the lifespan of C.elegans by
inducing autophagy via SIR-2.1 activation.
References[1]Yaguchi, Y.; Komura, T.; Kashima, N.; Tamura, M.; Kage-Nakadai, E.; Saeki, S.; Terao,
K.; Nishikawa, Y. Eur. J. Nut. 2014, 53, 1659.

45

P8
X-ray crystal structure analysis of the peripheral subunits-deleted
photosystem II for understanding of light-harvesting system
Yuri Kitashita1, Kouhei Ueda1, Keisuke Kawakami2, Yasufumi Umena2,3,7,
Masako Iwai4, Masahiko Ikeuchi5, Jian-Ren Shen6, Nobuo Kamiya1, 2
1
Graduate School of Science, Osaka City University, Osaka, Japan; 2The OCU Advanced
Research Institute for Natural Science & Technology (OCARINA); 3JST-PREST; 4Graduate
School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan;
5
Graduate School of Arts and Science, The University of Tokyo; 6Gradutate School of Natural
Science and Technology, Okayama University, Okayama, Japan; 7Present Address: Graduate
School of Natural Science and Technology, Okayama University, Okayama, JapanThe reaction center (P680) in photosystem II (PSII) of cyanobacteria and higher plants
absorbs solar light and performs charge separation. An electron produced is transferred from
P680 to a plastoquinone cofactor (QB) via the other one (QA). A positive charge remained at
P680 will extract one electron from the oxygen-evolving complex (OEC) in PSII. One oxygen
molecule is produced from two water molecules after four times of such kind of OEC
oxidation. PSII exists as a homodimer in the thylakoid membranes with molecular weight ofabout 700 kDa, and consists of the reaction center proteins (D1 and D2), the light-harvesting
antenna proteins (CP43 and CP47), three extrinsic subunits (PsbO, PsbU, PsbV), thirteen
low-molecular weight subunits (PsbE, PsbF, PsbH, PsbI, PsbJ, PsbK, PsbL, PsbM, PsbT,
PsbX, PsbY, PsbZ, and Psb30) and more than 100 cofactors such as chlorophylls andcarotenoids. PsbZ, one of the low-molecular weight subunits of PSII, is conserved fromcyanobacteria to higher plants. In Thermosynechococcus elongatus, a thermophilic
cyanobacterium, it has been reported that PsbK and Psb30 are deleted from purified PSII from
PsbZ-deletion mutant (ΔPsbZ-PSII) in its purification process. Additionally, PsbZ, PsbK and
Psb30 are located near CP43. The deletions of subunits induce structural changes of CP43,
and its structural changes may affect light-harvesting system of PSII. The aim of this study isto understand the light-harvesting system of PSII by functional and structural analyses of
ΔPsbZ-PSII. In order to investigate functional changes of ΔPsbZ-PSII, we performed
functional analyses of ΔPsbZ-PSII, in which the oxygen-evolving activity was lower in acomparison with wild-type PSII and the rate of the charge separation was decreased.
Moreover, we succeeded in collecting the diffraction data of ΔPsbZ-PSII crystal at aresolution of 2.46 Å. The space group of ΔPsbZ-PSII crystal is the same as that of wild-type
PSII crystal, but cell dimensions are different with each other. This indicates that theinteractions between PSII dimers related by crystallographic symmetries in ΔPsbZ-PSII
crystal are much different from wild-type PSII crystal. In this symposium, we will discuss thedetailed structural changes of ΔPsbZ-PSII.

46

P9Multi-photon excitation of photosystem II
by infrared femtosecond pulse laser
Kanako Nakata1, Yoshimasa Fukushima2, Keisuke Kawakami2,
Yasufumi Umena2, 3, 4, Tomoyuki Yatsuhashi1, 2, Nobuo Kamiya1, 2
1
Graduate School of Science, Osaka City University, Osaka, Japan; 2The OCU Advanced
Research Institute for Natural Science & Technology (OCARINA) , Osaka City University,
Osaka, Japan; 3JST-PREST; 4Present Affiliation: Graduate School of Natural Science andTechnology, Okayama University, Okayama, Japan
Multi-photon excitation is an electron transition process from ground state to excited stateaccompanying absorption of several photons at the same time. The multi-photon excitation by
femtosecond laser pulses has been utilized on various experimental situations such as
two-photon fluorescence-imaging microscopy. Photosystem II (PSII) exists in the thylakoid
membranes and contains chlorophyll a (Chl a) which is responsible for the energy conversion
from solar light into chemicals. Fluorescence of chlorophyll from the cells of photosyntheticorganisms by multi-photon excitation has been studied.
The purpose of this study is to elucidate the multi-photon optical characteristics of Chl a and
PSII. We irradiated infrared femtosecond laser pulses (wavelength 1320 nm, pulse width ~30
fs, frequency 5 kHz) to the solutions of Chl a (purchased from SIGMA-ALDRICH) in DMSO
and purified PSII, measuring fluorescence intensities and spectra. The fluorescence spectra
showed a peak around at 684 nm, indicating the emission from the S1 state (Qy band). The
number of photons absorbed by Chl a and PSII was estimated from the correlation between
the intensities of fluorescence and laser power. The slope in the double logarithmic plots were
3.0 for Chl a and 3.5 for PSII, respectively.
One-photon absorption spectrum of PSII
exhibits peaks at 436 nm (Soret) and 673 nm(Qy), respectively. Therefore, we concluded
at present that Chl a was excited to the S2(Soret) state with three photons of 1320 nm
laser pulses and the fluorescence was
emitted from the S1 (Qy) state. We will
discuss the multi-photon excitation processof Chl a (Figure1).

47

P10
Redox behavior of heterometallic trinuclear complexes
containing N-heterocyclic carbine platinum or palladium units
Yuri Maeda,1,* Hideki Hashimoto,1,2 and Takanori Nishioka1
1
Departments of Chemistry and Physics, Graduate School of Science, Osaka
City University, Osaka 558-8585, Japan; 2 The OCU Advanced Research Institute for Natural
Science and Technology, Osaka City University
Introduction
Multinuclear metal compounds are possible candidates of catalysts for multi-electron
processes because they can utilize their cluster valence electrons (CVE) and also provide
multiple reaction sites. For example, IrIII and RhIII trinuclear complexes with triply bridging
suldifes, [(MCp*)3(µ3-S)2]2+ (Cp* = η5-1,2,3,4,5-pentamethylcyclopentadienyl; M = Rh andIr), catalyze the electrochemical reduction of CO2 affording oxalate. There are only a few
examples of electrocatalysts that reduce CO2 to oxalate. For the tuning of properties of suchtrinuclear complexes, modification of Cp* ligands is required, however that is relatively hard
and limited, when compared to the N-heterocyclic carbene (NHC) ligand system. We planned
to replace one of the {MCp*} units in [(MCp*)3(µ3-S)2]2+ with a variety of chelated bisNHC
metal units and synthesized [(MCp*)2{M’(bisNHC)}(µ3-S)2](BPh4)2 (M’ = Pt, Pd).1 Reduced
species of the Pt complexes obtained by electrochemical reduction were also investigated.
Results and Discussion
The cyclic voltammograms of the palladium complexes showed two reversible reduction
peaks similar to the corresponding platinum complexes. Electrochemical reduction of theheterometallic trinuclear Rh2Pt complex wasinvestigated using UV-vis spectroscopy. The
electrochemically reduced monocationic andneutral Rh2Pt complexes showed absorption at
770 and 710 nm, respectively (Fig. 1) and theseabsorptions were assigned to SOMO-LUMO
and HOMO-LUMO transitions obtained fromthe results of DFT calculations. The
calculations for the 1e- and 2e-reduced and Fig. 1 UV-vis spectra for electrochemicallyoriginal complexes also exhibit the small reduced spices of the Rh2Pt complex.
structural changes upon the reductions
consistent with the results of the cyclic voltammetry showing two reversible reduction waves.
References
1. (a) Maeda et al. Inorg. Chem. 2014, 53, 661–663. (b) Maeda et al. Inorg. Chem. 2015, 54,
448–459.

48

P11
Syntheses and Catalytic Ability of Sugar-Incorporated
N-Heterocyclic Carbene Pincer Palladium Complexes    
Yosuke Imanaka,† Hideki Hashimoto, ‡,§ and Takanori Nishioka††
Department of Chemistry, Graduate School of Science, ‡Dkepartment of
Physics, Graduate School of Science, and § The OCU Advanced Research Institute for
Natural Science and Technology (OCARINA), Osaka City University, Osaka 558-8585, Japan
Introduction
Development of catalysts for organic reactions in water, which is a nontoxic andunexplosive liquid, has been studied for environmentally friendly chemical processes. For
such a purpose, we utilized click chemistry for incorporation of sugar units into transition
metal complexes affording water soluble complexes and reported synthesis of a C-C-N pincer
Pd complex containing a D-glucopyranosyl unit and its catalytic ability toward theSuzuki-Miyaura cross-coupling reaction in water. In
PF6
AcGlc = OAcNN
this study, we synthesized analogue complexes
R
O
AcO
AcO
N
OAc
possessing various N-substituents on the terminal
Pd
Cl
N[Pd(L-Me)Cl](PF6): R = MeNHC moiety of the tridentate ligands (Fig. 1), andN[Pd(L-iPr)Cl](PF6): R = iPr
N
[Pd(L-Bn)Cl](PF6): R = Bnevaluated the influence of the substituent on theirN
AcGlc [Pd(L-AcGlc)Cl](PF6) : R = AcGlc
structureandcatalytic
abilityfortheFig.1 Structure of the Pd complexes.
Suzuki-Miyaura cross-coupling reactions in water.
Results and Discussion
Table 1 Catalytic ability of Pd complexes
The tridentate ligand with an acetylated
in the Suzuki-Miyaura coupling reaction.
D-glucopyranosyl unit on the terminal NHC
moiety afforded only one of the diastereomersBr[Pd(L-R)Cl](PF6)
B(OH)2(1 mol ppm)of the Pd complex, which are derived from theK2CO3
twisted conformation of the ligand and thewater, 100 °C
16 hours
chiral D-glucopyranosyl unit, while the otherOOligands with achiral substituents on theR
Yield (%)
TON
terminal NHC units gave both of theMe
6.5
65,000diastereomers. The catalytic activity of Pd
iPr80
800,000
complexes toward the Suzuki-MiyauraBn
11110,000
cross-coupling reaction in water was
AcGlc
4.2
42,000examined (Table 1). [Pd(L-iPr)Cl](PF6)exhibited excellent catalytic activity withTON (800,000), which is comparable to the best value reported for a similar reaction. In
conclusion, we showed that modification of the N-substituents on the pincer ligands of the Pd
complexes is one of the effective methods to control their structures and catalytic ability.

49

P13
Structure and activity of
Mycoplasma mobile gliding machinery1

Miyuki Nishikawa, 2Daisuke Nakane, 3Akihiro Kawamoto,3
Takayuki Kato, 3,4Keiichi Namba, 1,5Makoto Miyata
1
Grad. Sch. of Sci., Osaka City Univ., 3-3-138, Sugimoto, Sumiyoshi-ku, Osaka, Osaka, Japan
2
Dept. Phys., Gakushuin Univ., 1-5-1, Toyoshima-ku, Tokyo, Japan
3
Grad. Sch. of Frontier Biosci., Osaka Univ., 1-3, Yamadaoka, Suita, Osaka, Japan
4
QBiC, RIKEN, 6-2-3, Furuedai, Suita, Osaka, Japan
5
OCARINA, Osaka City Univ., 3-3-138, Sugimoto, Sumiyoshi-ku, Osaka, Osaka, Japan
Mycoplasma mobile, a fish pathogen, glides on solid surfaces in the direction of a membrane
protrusion at a cell pole by a unique mechanism based on ATP energy [1-3]. The gliding
machinery is composed of surface huge proteins and internal “jellyfish structure” [4]. We
analyzed the gliding machinery three dimensionally by electron cryotomography, andelucidated that each cell has about 28 tentacles, which are aligned about 8 nm far from cell
membrane's internal surface. The tentacles were about 5 nm in diameter and 530 nm inlength decorated with globular particles of axes about 14 nm and 20 nm, in a pitch about 32
nm. We positioned the ATP binding sites by using fluorescently labeled ATP, MANT-ATP,and found that the tentacle part binds ATP. This result is consistent with the fact that theparalogs of α and β subunits of F1-ATPase are localized at the tentacle's position [5]. The
jellyfish structure hydrolyzed ATP with a rate 1 time per second per the β subunit paralog.
Next, we analyzed the structures in the presence of various ATP analogs through image
averaging of electron microscopy, and found that the globular particle is about 2 nm longer
under the conditions of AMPPNP, ADP-Vi, and ADP-AlFx than ADP and APO. These
structural variations may be involved in the gliding mechanism of M. mobile. We analyzed
the binding of isolated gliding machinery, “gliding head”, with ATP analogs. The binding
rate was twice under ADP condition than that under ATP condition. These result shows thatthe legs of M. mobile catches sialylated oligosaccharides fixed on solid surfaces under ADP
condition and releases that under ATP condition.
[1] Uenoyama, A.; Miyata, M. Proc Natl Acad Sci USA 2005, 102, 12754[2] Miyata, M. Annual Review of Microbiology 2010, 64, 519.
[3] Kinosita, Y.; Nakane, D.; Sugawa, M.; Masaike, T.; Mizutani, K.; Miyata, M.; Nishizaka,
T.      2014, 111, 8601
[4] Nakane, D.; Miyata, M.;      2007, 104, 19518[5] Tulum, I.; Yabe, M.; Uenoyama, A.; Miyata, M.      2014, 196, 1815

50

P13
Regulation of the ligand-binding ability of Zn-porphyrin with asterically-bulky shielding units by metal complexation
photo

Yoshikazu Ninomiya, Masatoshi Kozaki, Shuichi Suzuki, Keiji Okada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan

Abstract: Recently artificial allosteric systems have been attracting considerable interest
as smart catalysts, molecular machines, and targeted drug delivery reagents. We previously
found the Fe(II)-induced allosteric suppression for the ligand binding ability of the Zn
porphyrin unit in artificial allosteric system 1 with sterically bulky ligands [1]. We preparedartificial allosteric system 2 that expresses remarkable enhancement of the degree of allostericsuppression. Allosteric system 2 was synthesized according to the method similar to the reportof 1. UV-vis titration indicated the quantitative formation of Fe-complex 2•Fe (K2•Fe > 106
M 1) in toluene/acetonitrile (1/1 (v/v)). The degree of allosteric suppression for the binding of
3,5-bis(3,5-di-tert-butylphenyl)pyridine (L) with the Zn-porphyrin unit in 2 was estimated by
the comparison of the binding constants before and after the complexation. The binding−

constants K2•L and K(2•Fe)•L were determined to be 3.29 ± 0.10 × 104 M 1 and 0.348 ± 0.018 ×
104 M 1, respectively. The degree of the allosteric suppression (K2•L/ K(2•Fe)•L) was evaluatedas 9.5 times. The variable temperature titrations were carried out to obtain thermodynamic
parameters for the ligand binding reaction before and after the complexation. As a result, the−−reaction of 2•Fe and L has smaller enthalpy change (ΔH = −28 kJ mol 1) than the reaction of 2
and L (ΔH = −35 kJ mol 1), suggesting that the coordination bond between the nitrogen atomof L and the zinc atom of 2 in (2•Fe)•L is weakened due to steric congestion between
shielding units in 2 and L. [2]−−References:[1] Kozaki, M.; Ninomiya, Y.; Suzuki, S.; Okada, K. Tetrahedron Lett. 2013, 54, 3658.
[2] Ninomiya, Y.; Kozaki, M.; Suzuki, S.; Okada, K. Bull. Chem. Soc. Jpn. 2014, 87, 1195.

51

P14
Synthesis and properties of
2,11-dibromo-13,14-dimesityl-5,8-dioxapentaphene    
Chihiro SATO, Masatoshi KOZAKI, Shuichi SUZUKI, and Keiji OKADA
Graduate School of Science, Osaka City University,
3-3-138 Sugimoto, Sumiyoshi-ku, Osaka, Japan.
Abstract: Polycyclic conjugated systems with 4nπ electrons have received considerable
attention as organic-based electronic and optical materials due to their superior
electron-donating/accepting ability. Previously, we have investigated the unique redox
behaviors of electron-rich heteroacenes, such as phenothiazine or phenoxazine [1]. Here, we
designed and synthesized 5,8-dioxapentaphene delivative 1 as a novel electron-donating
system having 24π electrons.
Scheme 1. Synthesis of dioxapentaphene 1.

Dioxapentaphepe 1 was synthesized from 5-bromo-2-flurorbenzonitrile (2) in five steps(Scheme 1). The construction of a pentaphene-type framework was achieved byintramolecular cyclization of 4 using concentrated sulfuric acid. NMR and X-ray analyses
indicate that 1 has a considerably twisted π-framework
Br2
owing to steric repulsion between 13- and 14-mesityl groups(Figure 1). Despite the largely twisted structure, UV-vis
O1
spectrum of 1 in dichloromethane shows broad and intenseO2
absorption across the visible region (450–700 nm). Cyclic
voltammogram of 1 in dichloromethane reveals two Br1
reversible oxidation waves at +0.07 V and +0.42 V vs Fc/Fc+,
Figure 1. X-ray structure of 1.
which correspond to the generation of the cation and dication
species, respectively.
The detailed synthesis, photophysical and electrochemical properties of the compound will
be presented.
References: [1] Okamoto, T.; Kozaki, M.; Yamashita, T.; Okada, K. Tetrahedron Lett. 2001, 42, 7591.

52

P15
Induction of IL-8 secretion by bacterial flagellin in HEK293 cells and theinhibition by diffusely adherent Escherichia coli    
Takehiro Matuzaki1, Takumi Noju1, Sayaka Tamai2, Yoshihiko Tanimoto1,
Eriko Kage-Nakadai3, Toshio Kodama4, and Yosikazu Nishikawa1
1
Graduate School of Human Life Science, Osaka City Universi ,558-8585
2
Faculty School of Human Life Science, Osaka City University , 558-8585
3
The OCU Advanced Reserch Institute for Nature Science and Technology , 558-8585
4
Institute of Microbial Diseases, Osaka University , 565-0871[Background and aims ]Diarrheagenicity of diffulsely adherent Escherichia coli (DAEC) remains controversial
because of the prevalence not only among patients but healthy people. Previously, we foundthat all motile DAEC strains isolated from diarrheal patients induced high levels of IL-8
secretion in Caco-2 cells via toll-like receptor 5 (TLR5) [1-3]. In contrast, only 6 of 15 motile
strains from healthy carriers induced IL-8 secretion to the same levels despite their possessionof the flagella [4].
The DAEC that induced less IL-8 was referred to as MBLI (Motile-But-Low-Inducer)
strains. TLR5 is considered to be located on the basolateral site of intestinal epithelial cells.
We assumed that flagellin of MBLI strains could not reach TLR5 since MBLI could notloosen tight junctions as much as do the highly inducing strains from patients. However, there
was no significant difference between the epithelia infected by MBLI and those done by thehigh inducers [5]. The aim of this study is to elucidate the discrepancy in IL-8 inductionlevels among the DAEC strains.
[Materials and methods]
HEK293 cells that were transformed with TLR5-coding genes were infected by the DAEC
with or without the flagellin, TNF-α, or Phorbol 12-myristate 13-acetate (PMA). Then, IL-8
in culture supernatants were assayed by ELISA. The transcription levels of genes involved in
TLR signaling were assessed by RT-PCR Array (QIAGEN).
[Results and discussion]
When HEK293 cells were infected by the MBLI strains, none of flagellin, TNF-α and PMA
induced IL-8 secretion from the epithelial cells. This result indicates that MBLI strain
suppressed the response of epithelial cells to the inflammatory stimuli. Although the pathwaysof TNF-α and PMA to induce inflammatory reactions are different from that due to flagellin,
the MBLI strain could suppress the increase of IL-8 by the TNF-α or PMA stimulation. The
level of IL-8 mRNA transcription was increased regardless of which inflammatory stimulus
was used; MBLI had no effect on the siganal transduction and the mRNA transcription. These
data suggest that MBLI strains control the increase of IL-8 by suppression of the translation

53

P15
process or posttranslational events for the secretion rather than by inhibition of TLR5
signaling or mRNA transcription.
[1] Arikawa. K., Nishikawa.Y.
Microbiol. Immunol 2010, 54:491-501.
[2] Meraz. I. M., Arikawa.K.
Microbiol. Immunol 2006, 50:159-169.
[3] Arikawa. K., Meraz.I. Microbiol. Immunol 2005, 49:493-503.
[4] Fujihara. S., Arikawa.K.
Jpn. J. Infect. Dis 2009, 62:318-323.
[5] Tanimoto. Y., Arikawa.K. Vet. Immunol. Immunopathol 2013,152:183-188.

54

P16
Allosteric Control of the Axial-Binding Ability ofZinc Porphyrin Using a Stimulus Molecule    
SASAKI, Yoshito; KOZAKI, Masatoshi; SUZUKI Shuichi; OKADA, Keiji
Graduate School of Science, Osaka City University
Abstract: Artificial allosteric systems with metalloporphyrin units are attractive as intelligent
receptors and catalysts. Artificial allosteric system 1 was synthesized by introducing a
conjugated chain, which consists of a zinc diethynyldiphenylporphyrin center (ZnDEDPP)
and a diamidopyridine terminal (DAP), at 1 position on the benzene ring, and two side chains,which contains a zinc tetraphenylporphyrin terminals (ZnTPP), at 2,6-positions on thebenzene ring. A stimulus molecule 2 possessing a uracil (U) and two pyridine units (Py) was
also synthesized.
UV-vis spectra in toluene suggested that compounds 1 and 2 formed a tight 1 : 1 complex(log K > 8) through hydrogen bonds between DAP and U moieties and coordination bonds
between ZnTPP and Py moieties. The binding constant between ZnDEDPP in 1 and
4-phenylpyridine was 4.4 times larger than that between 1•2 and 4-phenylpyridine.
These results suggest that the formation of multiple supramolecular bonds result in thesignificant shielding of ZnDPDEP by side chains. We will present molecular design and theallosteric effect in detail.

Figure 1. Allosteric mechanism of compound 1 caused by stimulus molecule 2.

55

P17
  Three-dimensional mapping of groundwater geochemistry
related to the sources of groundwaters in Osaka Basin    

Tsuyoshi Shitani, Harue Masuda, Muneki Mitamura, Tatsuya Nemto andShinji Masumoto
Graduate School of Science, Osaka City Univeristy

Osaka Basin is a large reservoir of groundwater resources, which can be used for various
applications as industrial and domestic water resources. However, the uptake of groundwater
has been strictly regulated in the center of Osaka Basin since 1960s to avoid geogenic
disasters such as land subsidence, which actively occurred in the period of rapid economic
growth. The land subsidence has stopped since 1970s because of the regulation, andgroundwater has not been extensively used for more than three decades. However, the uptakeof groundwater has been a threat again due to increasing consumption of groundwater for
private water supplies since 2000s. Depths of private wells for industries, hospitals, etc., aremostly from 100 to 300m from the ground surface, where the shrinking clay layers severely
occurred. Because those groundwaters have not been used for long time, present waterchemistry is not well documented.
In this study, groundwaters mainly sampled from the wells between 100 and 300m depths
were studied for the stable hydrogen and oxygen isotope ratios and major chemicalcomponents to estimate the origins of groundwaters. Combining the results of this study andprevious studies, three-dimensional mapping of groundwater geochemistry was drawn to
discuss the groundwater flow system of the basin and the effect of the land subsidence to thepresent groundwater geochemistry. Aquifers of groundwater were classified by geologicalinformation including marine clay layers. This study was successful to visualize thegroundwater chemistry as three-dimensional maps, which clearly show the following featuresof groundwater chemistry.
Hydrogen and oxygen isotope ratios of the groundwater 100m depths increased frommountainous areas to the center of plain, and the origin of these groundwaters were local
meteoric water. In the western plain of Uemachi plateau, of which altitude is below sea level,
the stable isotope ratios of groundwaters (δ2H: -40~, δ18O: -5~) are larger than those of
local meteoric water (δ2H: -45~-40, δ18O: -7~-6), due to mixing with seawater. Thus,
the seawater invaded into the aquifers of these areas, especially those between marine clay
layers Ma12 and Ma9.
The stable isotope ratios of the groundwater were low (δ2H: ~-55, δ18O: ~-8) in the
deeper aquifers than the Ma9. Slightly lower isotope ratios of the groundwaters than those of
local meteoric water with diluted Na-HCO3 type chemistry suggested that the groundwatercontained squeezed pore water from the overlying clay layers. It would be the evidence ofexcessive wage of groundwater when the land subsidence actively occurred.

56

P18
Optimization of crystallization conditions
for photosystem II complex
from thermophilic cyanobacteria
Naoto Inohara1, Keisuke Kawakami2, Nobuo Kamiya1, 2
1
Graduate School of Science, Osaka City University, Osaka, Japan, 2TheOCU Advanced Research Institute for Natural Science & Technology (OCARINA), Osaka
City University, Osaka, Japan.
Crystal structure of water-splitting and oxygen-evolving photosystem II (PSII) from
Thermosynechococcus vulcanus has been reported at a resolution of 1.9 Å using X-rays from asynchrotron facility, SPring-8 [1]. PSII absorbs solar light at the reaction center, P680, to
produce a pair of an electron and a hole. The electron is transferred to reduction side of PSII
and the hole is compensated by an electron extracted from oxygen-evolving complex (OEC),
the Mn4CaO5 cluster. Oxygen evolution undergoes at OEC according to the Kok cycle modelthrough five intermediate states Si (i=0, 4) with different oxidataion equivalences. In order toelucidate the Kok cycle mechanism, to obtain PSII crystals of much higher quality isnecessary to analyse the intermediate structures of OEC at much higher resolution.
In this study, we performed screening of PSII crystallization conditions to realize the highquality PSII crystals using several kinds of detergents. HTG (n-heptyl-β-D-thio glucoside) is
one of the detergents used for purification and crystallization of membrane proteins. Chemical
structures of three detergents (HECAMEG, methyl-6-O-(N-heptylcarbamoyl)-α-D-Gluco
pyranoside; β-OG, n-Octyl-β-D-glucopyranoside; OTG, n-Octyl-β-D-thioglucopyranoside)
are similar to that of HTG. In addition, we used two detergents (α-DDM,
Dodecyl-α-D-maltoside; SM, Sucrose monolaurate) in crystallization, having similar
chemical structures to β-DDM (Dodecyl-β-D-maltoside) which was used for purification of
PSII. X-ray diffraction patterns from PSII crystals were recorded by a laboratory X-ray source(RIGAKU R-AXIS VII) at ReCAP (Research Center for Artificial Photosynthesis). The
diffraction data and mosaicities were calculated using XDS (X-ray Diffraction Software) andHKL2000 (HKL Research Inc. USA), respectively. PSII crystals treated with HTG,HECAMEG, and β-OG diffracted to 2.7 Å, 2.7 Å and 2.8 Å resolution, respectively. Mosaicities
of HECAMEG- and OG-treated crystals were ~0.6° which were smaller than that of
HTG-treated crystal (~1.0°). On the other hand, No sopt was observed in the diffraction
patterns of α-DDM- and SM-treated crystals. Furthermore, electron transfer activitiies of PSII,
CMC (Critical Micelle Concentration) values of detergents, and hydrodynamic radii of PSII
were measured in crystallization and/or cryoprotectant solutions using fluorescence
spectrophotometer and DLS (Dynamic Light Scattering) device, respectively.
[1] Umena Y., Kawakami K., Shen J.R., Kamiya N., Nature, 473, 55-60 (2011).

57

P19
Molecular mechanisms underlying immunosenescenceto legionella infection in Caenorhabditis elegans    
Yuuya Hinode1, Tomomi Komura1, Eriko Kage-Nakadai1,2, Yoshikazu Nishikawa1
1
Graduate School of Human Life Science; 2 OCU Advanced Research Institute of Natural
Sciences and Technology, Osaka City University, 558-8585, Japan[Introduction] Legionella pneumophila (LP) is Gram-negative bacilli and a humanopportunistic pathogen. We previously found that young Caenorhabditis elegans animals(3-day-old) were resistant to LP, but old animals (8-day-old) were susceptible to the LP
infection, suggesting that C.elegans is useful as a model of opportunistic infection with
LP[1,2]. In this study, we investigated the molecular mechanisms underlying
immunosenescence of C.elegans to LP.
[Methods] C.elegans animals synchronized by an egg preparation were fed Escherichia coli
OP50 (OP) until they reached 3-day-old adults or 8-days-old adults. Then, the worms werefed OP or LP, and the lifespan was measured. Transcriptional changes elicited 24h after the
LP infections were determined by the microarray analysis and qRT-PCR. Lipid accumulation
was examined by Oil Red O staining.
[Result and Discussion] Consistent with our previous report, there were no significant
differences in survival between 3-day-old animals fed OP and those infected with LP. On theother hand, 8-day-old worms infected with LP exhibited significantly shortened lifespan whencompared to worms fed OP. In addition, pmk-1 /p38 MAPK mutants showed shortened
lifespan than the wild type nevertheless they were infected from 3-day-old stages. These
results suggested that the p38 MAPK pathway was involved in the host defense to the LP
infection. Microarray and qRT-PCR analyses revealed that many lipase genes wereupregulated by LP infection specifically in the 3-day-old infections, while no or slightchanges were observed in the worms infected with LP at the 8-day-old. Oil Red O staining
confirmed reduced lipid accumulation in the young worms infected with LP. These results
suggest that lipase genes may be involved in the resistance LP in the young animals under thecontrol of pmk-1. We have been investigating functions of genes, which were upregulated by
LP infection, using the mutants and RNA interference.
References[1] Komura, T., Yasui, C., Miyamoto, H., Nishikawa, Y. Appl. Environ. Microbiol.
2010;76:4105-4108.
[2] Komura T, Ikeda T, Hoshino K, Shibamura A, Nishikawa Y. Adv Exp Med Biol.
2012;710:19-27. doi: 10.1007/978-1-4419-5638-5_3.

58

P20
Biased gliding direction of Mycoplasma mobile
Hanako MORIO, Taishi KASAI, Makoto MIYATA
1) Graduate School of Science, Osaka City University
2) OCARINA, Osaka City University
Abstract: Mycoplasma mobile forms a membrane protrusion at a cell pole and glides in its
direction by a unique mechanism in which mycoplasma legs catch, pull, and release sialylated
oligosaccharides fixed on a solid surface [1, 2]. In this study, we analyzed the gliding
direction and found that 77% cells glided to the left with changing the direction for -8.4 ±17.6 degrees per 1 µm displacement. To clarify this feature based on the putative gliding
mechanism, we focused on the whole cell movement. A 200 nm bead attached to a cellbody did not roll around the cell axis. Cells elongated by the addition of detergent showedlarger change in the gliding direction. These results suggest that the propelling force biased
from the cell axis drags the back side, resulting in the change in the gliding direction. Next,
we focused on leg movements. Under high viscosity conditions containing methylcellulose
or Ficoll, the distribution of gliding direction decreased, suggesting that the change in gliding
direction is based on the structure of gliding machinery. In the presence of 250 µM freesialylated oligosaccharides, in which the binding of leg is reduced, 70% and 30% cells glided
to the left and the right with changes of 30 degrees per 1 µm. The gliding ghosts
permeabilized and by Triton X-100 and reactivated by ATP showed no change in the gliding
directions, probably because the treatment by detergent damaged the unit alignment of gliding
machinery. The results obtained here may be explained based on an assumption below.
The units of gliding machinery are aligned helically around the machinery. The legs catchthe sialylated oligosaccharides after some wobble and pull the cell, and generate thepropelling force tilted from the cell axis, and the 70% propelling force work to the left, and
30% work to the right. The drag generated in the back side induces the change in gliding
direction.
References:[1] Miyata M (2010) Unique centipede mechanism of Mycoplasma gliding. Annual Review ofMicrobiology 64: 519-537.
[2] Miyata M, Nakane D (2013) Gliding mechanism of Mycoplasma pneumoniae
subgroup implication from Mycoplasma mobile. In: Browning G, Citti C,
editors. Molecular and cell biology of Mollicutes. Norfolk, UK: Horizon
Press. pp. 237-252.

59

P21
Development of Catalytic Asymmetric Diels−Alder Reaction
with Nitro Olefin Using Thiourea Catalyst    
Shota Ueda, Atsushi Manabe, and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Diels−Alder (DA) reaction is a powerful synthetic method for the construction ofhighly substituted cyclohexene rings. In this presentation, we would like to report our
synthetic efforts toward development of chiral organocatalyst-mediated asymmetric DA
reaction using β-nitro acrylate 1 and the electron-rich furan 2. To the best of our knowledge,
the asymmetric DA reaction of 1 and 2 has not been examined yet.
DA reaction of 1 and 2 in the presence of thiourea catalyst A proceeded smoothly toafford the desired DA product 3 as a hemiaminal form along with the conjugate addition
product 4 in low enantioselectivities. After extensive studies, we found that selectivity (3 vs4) was improved by using catalyst C in toluene to give 3 in 81% ee.
NO2

EtO2C

O
NHBoc

1

N

O2N

CO2Et
O

O

+

CO2Et

OH
NHBoc

solvent, −40 oC, 24 h

2

NHBoc
4

3

entry

catalyst

solvent

3

4

1

A

CH2Cl2

12% (2% ee)

28% (7% ee)2

BCH2Cl238% (40% ee)

9% (52% ee)

3C

CH2Cl2>59% (58% ee)

18% (59% ee)

4Ctoluene38% (81% ee)

15% (72% ee)

N

CF3

S

N
H

N
H

CF3

Bn

CF3

t-Bu
S

N
O

N
H

N
H

Bn
CF3

t-Bu
S

N
O

N
H

N
H

B

N

HO
t-Bu

A

60

catalyst (5 mol%)

+

O2N

C

t-Bu

P22
Development of Caspase-3 Inhibitors Using Squaric Acid Derivatives    
Kentaro Maeda and Tetsuro Shinada
Graduate School of Science, Osaka City University
3-3-138 Sugimoto, Sumiyoshi, Osaka 558-8585, Japan
Caspases are a family of cysteine proteases that play the key roles in signaltransduction cascades in apoptosis and inflammation. Excessive neuronal apoptosis leads toneurodegenerative disorders such as stroke, Alzheimer’s disease, Huntington’s disease andParkinson’s disease. Therefore, caspases are recognized as therapeutic targets for these central
nervous diseases.
Recently, we have developed α-amino squaric acid (α-Asq) 1 as a novel amino acidanalog.[1] The squaric moiety have an electron deficient property due to the presence of thetwo continuous carbonyl groups. Indeed, t-butyl methyl squarate readily reacted with thiols togive the corresponding addition-elimination products.[2] We hypothesised that the squaric
moiety of 2 and 3 would react with the active site thiols of caspases to be the competitive
inhibitors. In this presentation, the synthetic details and biological evaluations of 2 and 3 will
be reported.

[1] (a) Ishida, T.; Shinada, T.; Ohfune, Y. Tetrahedron Lett. 2005, 46, 311-314.(b) Ishida, T.;
Shinada, T.; Ohfune, Y. Synthesis 2005, 2723-2729.
[2] Shinada, T.; Yamasaki, A.; Kiniwa, Y.; Shimamoto, K.; Ohfune, Y. Tetrahedron Lett.
2007, 48, 7614-7617.

61

P23
Structural analysis of P65 protein essential for
Mycoplasma pneumoniae gliding
Masaru Yabe1, Miki Kinoshita1,
Katsumi Imada3, Yukio Furukawa4,
Munehito Arai5 and Makoto Miyata1,2
1 Graduate School of Science, Osaka City University
2 OCARINA, Osaka City University
3 Graduate School of Science, Osaka University
4 Graduate School of Frontier Bioscience, Osaka University
5 Graduate School of Arts and Sciences, The University of Tokyo
Mycoplasma pneumoniae, a human pathogen, glides to one direction with a uniquemechanism. The gliding machinery localizing at a cell pole contains an internal structurecomposed of terminal button, paired plates, and bowl complex from the distal end [1-2]. In
the present study, We focus on P65 (MPN309 protein), localizing at the terminal button. We
expressed recombinant P65 in E. coli and purified. The negative staining electron microscopy
showed that the protein forms a multimer composed of a central globule 22 nm in diameterand many filaments about 15 nm sticking out, consistent with the image of the terminal button
observed in a cell. The partial digestion by trypsin showed that 1-209 amino acid region(Fragment 1) and 212-283 amino acid region (Fragment 2) are stable. Intrinsically disorderedregion prediction, analytical ultracentrifugation, circular dichroism (CD) spectroscopy, small
angle X-ray light scattering results showed that Fragment1 is disordered filamentous
monomer. Size-exclusion chromatography-multi angle light scattering showed that Fragent 2
forms a pentamer. We obtained rod-shaped crystals of Fragment 2 that diffracts up to 2.7 Å
resolution and constructed an electron density map. Based on these results, We concluded that212-283 amino acid region is involved in pentamerization and 284-405 amino acid region
forms a multimer composed of a central globule and many filaments sticking out, whichinteract with other gliding proteins involved in gliding mechanism.
[1] Miyata, M., Annu Revew of Microbiology, 2010, 64:519-537.
[2] Miyata, M, and Nakane D., 2013, Molecular and cell biology of Mollicutes, 2013,237-252.

62

P24
Synthesis and Magnetic Properties of Tris[Gold(I)-(Imino
Nitroxide)-2-ide Radical Anion] Complexes
Tomoyuki Wada, Shuichi Suzuki, Masatoshi Kozaki, Daisuke Shiomi,
Kazunobu Sato, Takeji Takui, Keiji Okada
Department of Chemistry, Graduate School of Science, Osaka City University, Sugimoto,Sumiyoshi-ku, Osaka 558-8585, Japan.
Recently, we have developed metal-radical complexes using(nitronyl nitroxide)-2-ide radical (NN) anion as a spin-possessing
ligand and investigated their electronic and magnetic susceptibility[1−3]. Now, we succeeded in synthesizing C3 symmetrictris[gold(I)-(imino nitroxide)-2-ide radical anion] complexes. The
trinuclear complex 1 was prepared accoding to Scheme 1:
Chloro(triphenylphosphine)gold 2 was treated with nitronyl
nitroxide in the presence of NaOtBu in CH2Cl2 to give
NNAu(I)PPh3 (3), which was converted to trinuclear
complex 1 by the treatment with sodium nitrite in acidic
condition. Crystals suitable for X-ray structure analysis wereobtainedbyrecrystallized

frombenzene/cyclohexane;

trinuclear complex 1 has a shallow bowl-like structure andmakes a short contact to form an intermolecular dimeric
structure (Figure 1). The distance between three gold planes

Figure 1. Dimeric structureof trinuclear complex 1.

is 3.34 Å. When trinuclear
complex 1 was treated
with

0.5

equivalentamount of AgOTf, the Ag+
ion was inserted between
the dimeric forms. In thiscontribution,

we

will

discuss in detail described above.
References:[1] Okada, K. et al. J. Am. Chem. Soc. 2012, 134, 17866−17868.
[2] Okada, K. et al. Chem. Lett. 2014, 43, 678.
[3] Okada, K. et al. Eur. J. Inorg. Chem. 2014, 4740−4744.

63

P25
Surface Gliding Proteins of Mycoplasma mobile
Visualized by Quick-Freeze Deep-Etch Replica
Electron Microscopy    
Clothilde BERTIN1,2, Yuhei O TAHARA1,2, Eisaku KATAYAMA1,2, Makoto MIYATA1,2
1) Graduate School of Science, Osaka City University
2) OCARINA, Osaka City Universit
Mycoplasma mobile, the fatest gliding mycoplasma, belongs to the Mollicutes, aclass of wall less and small bacteria [1]. Dozens of mycoplasmas can glide on solid surfaces
including their host cells trough a unique mechanism. The gliding machinery of M. mobile
relies notably on “leg” proteins which propel the cells by repeated catching, pulling andreleasing sialylated oligosaccharides (SOs) and crank proteins transmiting the gliding force
provided by ATP hydrolysis [2]. In the present work, we tried to visualize the gliding
machinery structures by quick-freeze deep-etch replica electron microscopy (QFDE-EM).
QFDE-EM offers a high spatial resolution for the three-dimentional visualization of isolated
proteins, cells and biological processes [3]. In order to freeze the cells during gliding and toincrease resolution, washed cells were put on mica flakes [4] coated with extensively
fragmented fetuin, a source of SOs. In parallel, cells were submitted to an osmotic shock inorder to access to hidden structures.
These techniques allowed us to visualize leg structures, 73 nm in total length, divided
into two parts: a rod sticking out from the cell surface ended by a globular part (Fig. 1). The
dimensions of legs are consistent with those of isolated leg protein [5].

Visualization ofM.
mobilemutant,deficient in theexpressi
on of anessentia
l gliding protein, revealed a novel "starfish" structure which might correspond to the crankprotein assembly (Fig. 2). Moreover, the first analysis showed that crank protein would form

64

P25
a triskelion structure as observed previously with single protein experiments [6].These results
are the first step to understand the gliding mechanism of M. mobile by the vizualisation ofprotein assembly on frozen living cells.

[1] Razin, S. & Hayflick, L. Biologicals : Journal of the International Association of
Biological Standardization, 2010, 38, 2, 183.
[2] Miyata, M. Annual Review of Microbiology, 2010, 64, 519.
[3] Heuser, J. Trends in Biochemical Sciences, 1981, 6, 3, 64.
[4] Katayama, E. Journal of Molecular Biology, 1998, 278, 2, 349.
[5] Adan-Kubo, J. Journal of Bacteriology, 2006, 188, 8, 2821.
[6] Nonaka, T., Adan-Kubo, J. Journal of Bacteriology, 2010, 192, 3, 636.

65

P26
Structurale analyseis of Mycoplasma mobile gliding protein
by electron microscopy and high speed atomic force microscopy
Yuhei Tahara1, Noriyuki Kodera3, Toshio Ando3, Makoto Miyata1,21)Graduate School of Science, Osaka City University2)OCARINA, Osaka City University3)Bio-AFM Frontier Research Center, Kanazawa University
Mycoplasma mobile, a fish pathogen, glides on solid surfaces with a unique mechanism.
The gliding machinery is assembled on the protrusion mainly from two huge proteins,
including Gli521 and Gli349, weighing 521k, 349k, respectively[1]. In this study, we
analyzed molecular shapes of these proteins essential for gliding, by negative-staining
electron microscopy and high speed AFM[2]. Gli521 has the role of “crank”, transmitting
movements from motor to leg. Previously, rotary shadowing electron microscopy showedthat the Gli521 molecule is 120 nm-“ ? ” shape, featured with a “hook” at the C terminus andan “oval” at the N terminus. In the present study, we showed that the molecule is shaped“ω”, featured with a central hinge, based on better resolution of negative–staining electron
mycroscopiy. High speed AFM showed that the central hinge bends only to one direction.
Gli349, has been shown to be 100 nm-“ ♪ “ shape and function as “leg”. High speed AFM
showed that the Gli349 molecule consists of a large and a small domains connected by a thinelastic filament. The results gave us better images of the gliding machinery of M. mobile.
[1] Miyata. M., Annual Review of Microbiology, 2010, 64, 519.
[2] Ando. T., Uchihashi. T., Kodera. N., Yamamoto. D., Miyagi. A., Taniguchi. M., andYamashita. H. 2008, Proc Natl Acad Sci U S A 98, 12468.
[3] Adan-Kubo. J., Uenoyama. A., Arata. T., Miyata. M., J. Journal of Bacteriology,
2006,188,2821[4] Nonaka. N., Adan-Kubo. J., Miyata. M., J. Journal of Bacteriology, 2010, 192, 636

66

P27
3D structural analysis of the crucial Intermediateof skeletal muscle myosin and its role
in revised actomyosin cross-bridge cycle    
Eisaku KATAYAMA
Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku,
Osaka-shi, Osaka 558-8585, Japan
Abstract: Skeletal myosin S1 consists of two functional segments, a catalytic-domain and alever-arm. Since the crystal structure of ADP/Vi-bound S1 exhibits a strong intramolecular
flexure between two segments, inter-conversion between bent and extended forms; i.e. “tiltingof the lever-arm” has been accepted as the established molecular mechanism of skeletal
muscle contraction [1]. We utilized quick-freeze deep-etch replica electron microscopy to
directly visualize the structure of in vitro actin-sliding myosin [2], and found the existence ofa novel oppositely-bent configuration, instead of the expected ADP/Vi-bound form. We also
noticed that SH1–SH2 cross-linked myosin gives an aberrant appearance similar to the above
structure [3]. Since SH1–SH2-cross-linked myosin is a well-studied analogue of thetransient intermediate of the actomyosin cross-bridge cycle, we devised a newimage-processing procedure to define the relative view-angles between the catalytic-domain
and the lever-arm from those averaged images, and built a 3-D model of the new conformer[4, 5]. The lever-arm in that model was bent oppositely to the ADP/Vi-bound form, in
accordance with observed actin-sliding cross-bridge structure. Introducing this conformer as
the crucial intermediate that transiently appears during sliding, we propose a revised schemeof the cross-bridge cycle [5, 7]. In the scenario, the novel conformer keeps actin-binding in
two different modes until it forms a primed configuration. The final extension of thelever-arm back to the original rigor-state constitutes the “power-stroke”. Various images
observed during sliding could be easily interpreted by the new conformer. Even the enigmatic
behavior of the cross-bridges reported as “loose chemo-mechanical coupling” [6] might beadequately explained under some assumptions [7].
References:[1] Vale, R. D. and Milligan, R. A. Science 2000, 288, 88.
[2] Katayama, E. J. Mol. Biol. 1998, 278, 349[3] Katayama, E.et al. Adv. Exp. Med. Biol. 2003, 538, 295.
[4] Kimori, Y. et al. Ultramicroscopy 2007, 107, 25.
[5] Kimori, Y., Baba, N. and Katayama, E. Biochem. J. 2013, 450, 23.
[6] Cyranoski, D. Swimming against the tide. Nature 2000, 408, 764.
[7] Katayama, E. BIOPHYSICS 2014, 10, 89

67

P28
Localization and roles of F1-ATPase subunit paralogs
of Mycoplasma mobile revealed by gene manipulation
Isil Tulum1, Kenta Kimura1, Masaru Yabe1, Atsuko Uenoyama1 and Makoto Miyata1,21) Graduate School of Science, Osaka City University; 2) OCARINA , Osaka City University
Mycoplasmas are parasitic and occasionally commensal bacteria that lack peptidoglycan
layers and have small genomes. Mycoplasma mobile, a fish pathogen, has a membrane
protrusion at one pole and exhibits a unique gliding motility in the direction of the protrusion.
The gliding machinery is composed of surface and internal structures. The internal "thejellyfish structure" is composed of "bell", the front and "tentacles", the end parts. The energy
for motility is supplied by ATP [1,2].
In the present study, we developed a method for fluorescent protein tagging to determine thesubcellular localization of jellyfish proteins and Gli42, tubulin-like protein. We found that
Gli42 localizes at the gliding machinery, and a paralog of the F1-ATPase α-subunit, together
with a paralog of the F1-ATPase β-subunit, forms the tentacles of the jellyfish structure [3].
This may suggest that the movement for gliding is generated by the internal structure.
Interestingly, the jellyfish structure genes have high similarity to the original copy of
FoF1-ATPase operon. In order to prove this assumption, we are* doing EYFP tagging of other jellyfish proteins.
* applying site specific mutagenesis to detect the ATP hydrolysis function of these proteins.
* creating a new model transferring the whole jellyfish structure into the cell wall-lessBacillus, L-form.

References[1] Miyata, M. Annual Review of Microbiology, 2010, 64, 519.
[2] Kinosita Y, Nakane D, Sugawa M, Masaike T, Mizutani K, Miyata M, Nishizaka T.
PNAS.2014, 111, 8601.
[3] Tulum, I, Masaru, Y, Uenoyama, A, and Miyata, M. Journal of Bacteriology, 2014,
196(10), 1815.

68

P29
Gliding motility of Mycoplasma mobile
on uniform sialylated oligosaccharide
Taishi KASAI1, Tasuku HAMAGUCHI1 and Makoto MIYATA1,2
1) Graduate school of science, Osaka city university
2) OCARINA, Osaka city university
Some Mycoplasma species bind to solid surfaces and glide to one direction. Usually,
mycoplasmas glide on mixed sialylated oligosaccharides (SOs) derived from glycoprotein andglycolipid, and gliding motility on uniform SOs has never been observed. In the present
study, we analyzed the gliding of M. mobile on a plastic plate covered by one of 53 uniform
oligosaccharides. Mycoplasmas bound to and glided on only 21 of the fixed SOs, showingthat sialic acid is essential as the binding target. The affinities were mostly consistent with
our previous results on the inhibitory effects of free SOs [1,2]. The results suggest that therecognition mechanism of SOs by M. mobile are as follows: (i) the nonreducing terminal
Neu5Ac is tightly recognized in an occluded position, and (ii) Branching oligosaccharide are
well recognized with the position of branching, sort of branching sugar and orientation ofeach linkage. The cells glided on uniform SOs in manners apparently similar to those of thegliding on mixed SOs. The gliding speed was related inversely to mycoplasma’s affinity forSO, suggesting that the detaching step may be one of the speed determinants. The cells
glided faster and with smaller fluctuation on the uniform SOs than on the mixtures,
suggesting that the drag caused by the variety in SOs influences gliding behaviors.
[1] Kasai, T.; Nakane, D.; Ishida, H.; Ando, H.; Kiso, M.; Miyata, M.; Journalof Bacteriology
2012, 195, 425.
[2] Kasai, T.; Nakane, D.; Ishida, H.; Ando, H.; Kiso, M.; Miyata, M.; Bio-protocol 2013, 3

69

P30
Redox properties of incomplete cubane type
Mo3X4 (X = S or O) clusters    †
Keisuke Kawamoto, †Akio Ichimura, †,$Hideki Hashimoto, †,$Isamu Kinoshita,†
Masakazu Hirotsu, †Takanori Nishioka†
Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka,558-8585, Japan (keisuke@sci.osaka-cu.ac.jp); $The OCU Advanced Research Institute for
Natural Science and Technology (OCARINA), Osaka City University, 3-3-138 Sugimoto,Sumiyoshi-ku, Osaka, 558-8585, Japan.
Abstract: Nanoparticle of MoS2 has been widely attracted much attention as an excellent
catalyst for hydrogen evolution reaction (HER) [1]. Complexes of the Mo3S4 framework(Chart 1) have also been reported as a catalyst for HER. However, the mechanism of thecatalytic HER by these compounds has not been clarified yet. It is noteworthy that the Mo3S4
complexes have an incomplete cuboidal structure. The bridged µ2-S ligands incorporate
various metals or organic compounds because an unoccupied molecular orbital on the µ2-S isthe main component of LUMO [2].
We have previously reported the synthesis and the redox Chart 1. Structure of the
Mo3S4 framework.
properties of Mo3S4 complexes with Schiff base ligands as afundamental framework for investigating the catalytic mechanism
for the proton reduction reaction. These investigations revealed
that the catalytic proton reduction reaction occurs on the bridgedµ2-S ligands of the Mo3S4 core [3].
  In the isolated form of the Mo3S4 cluster, all molybdenum
centers have tetra-valent oxidation states. The above reactions such as the catalytic proton
reduction reaction and the incorporation reaction of metal ion are induced by the reduction
from Mo(IV) to Mo(III). On the other hand, we are also interested in the properties of Mo(V)centers on the Mo3S4 core because some biological systems containing molybdenum center
utilize a redox reaction between Mo(VI) and Mo(IV). With this background in mind, it is very
important to discuss the properties of the Mo3S4 clusters in the redox processes. Therefore, inthis conference, we will discuss the stabilities of the mixed-valent Mo3S4 clusters containingsome oxidation numbers of Mo(III), Mo(IV), and Mo(V) [4].
References: [1] Kibsgaard, J.; Jaramillo, T. F.; Besenbacher, F. Nature Chem. 2014, 6,
248-253. [2] Shibahara, T.; Kawamoto, K.; Matsuura, A.; Takagi, H.; Nishioka, T.; Kinoshita,
I.; Akashi, H. Bull. Chem. Soc. Jpn. 2014, 87, 459-469. [3] Kawamoto, K.; Ichimura, A.;
Hashimoto, H.; Kinoshita, I.; Nishioka, T. Bull. Chem. Soc. Jpn., 2015, 88, DOI:
10.1246/bcsj.20140275. [4] Kawamoto, K.; Ichimura, A.; Hashimoto, H.; Kinoshita, I.;
Hirotsu, M.; Nishioka, T. Bull. Chem. Soc. Jpn. 2015, 88, DOI:10.1246/bcsj.20140364.

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Effect of pgsE on microbial production of poly γ-glutamic acid
–Estimation of molecular weight based on intrinsic viscosity and
SEC-MALS analyses1

Takashi Tomiyama, 1Takahiro Inoi, 2Takashi Nishiyama, 2Hidetoshi Horibe, 3Ryosuke
Takahashi, 3Shinichi Kitamura, 4,5Yasufumi Umena, 1,6Akira Ogita, 1Toshio Tanaka,1
Ken-ichi Fujita
1
Graduate School of Science, Osaka City University,
2
Graduate School of Engineering, Osaka City University,
3
Graduate School of Life and Environmental Sciences, Osaka Prefecture University,
Gakuencho Nakaku, Sakai 599-8531, Japan,
4
OCU Advanced Research Institute for Natural Science and Technology,

PRESTO, Japan Science and Technology Agency,Research Center for Urban Health and Sports, Osaka City University

Poly γ-glutamic acid (γ-PGA) is one of biopolymers produced by Bacillus sp. strains.
γ-PGA is expected to be applied to many manufacturing products because of its uniquefunctions: water-absorbing, moisture-retaining and immunostimulating properties, etc [1].
These properties are potentiated as increasing the molecular weight (MW) of the polymer.
However, the biosynthetic mechanism of γ-PGA has been unclear until now. In addition, theproduction ablity of high MW of γ-PGAs is unstable in most of Bacillus sp. Strains.
Pgs operon for γ-PGA biosynthesis is composed of 4 genes, namely, pgsB, pgsC, pgsA,and pgsE. The 3 genes, pgsB, pgsC, and pgsA has been reported to be essential to the
production of γ-PGA [2]. Previous study suggested that pgsE may be involeved in γ-PGA
production ablity. To further understand the function of pgsE, we compared the MW of
γ-PGA produced in cells with or without pgsE expression based on intrinsic viscosity and
SEC-MALS analyses [3].
Bacillus sp. F-2-01 (F-2-01) shows high productivity of γ-PGA [4], and the MW of
γ-PGA derived from F-2-01 strain is larger than that from other γ-PGA producing strains. A
type strain of B. subutilis 168 (168) shows nonproductition of γ-PGA. We constructed cellswith high expression systems using pgsBCA or pgsBCAE derived from F-2-01 strain in
γ-PGA-nonproducing 168 strain.
From the results of dynamic light scattering, the size distribution of γ-PGA produced bycells with or without pgsE was approximately 10-100 or 80-200 nm, respectively. Based ondata obtained from intrinsic viscosity, the MW of γ-PGA in the cells with or without pgsE
was estimated as approximately 670,000 or 4,400,000, respectively. Based on analysis using
SEC-MALS, the MW of γ-PGA was estimated as approximately 4,500,000 in the cells with
pgsE. While, SEC-MALS analysis revealed that a purified sample obtained from the cells
without pgsE contained γ-PGA of MW 900,000 in small amounts and mainly did unidentified
materials of MW 28,000 and 6000. Which of the unidentified materials possively is the main

71

P31
constituent of γ-PGA. Thus, the results indicated that the MW of γ-PGA is probably smaller
than that previously expected in the cells without pgsE.
These findings suggested that pgsE contributes to an increase in the MW in addition to the
productivity of γ-PGA.
[1] Thomas, C., Agnes F, Molecular Microbiology 2006, 60(5), 1091-1098.
[2] Ashiuchi, M., Shimanouchi, K., Horiuchi, T., Kamei, T., Misono, H. Biosci. Biotechnol.
Biochem. 2006, 70(7):1794-1797.
[3] Irurzun, I., Bou, J. J., Pèrez-Camero, G., Abad, C., Campos, A., Muæoz-Guerra, S.,
Macromol. Chem. Phys., 2001, 202, 3253–3256.
[4] Kubota, H., Matsunobu, T., Uotani, K., Takebe, H., Satoh, A., Tanaka, T., Taniguchi, M.,
Biosci. Biotech. Biochem., 1993 57(7), 1212-1213.

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Design of novel PG-surfactants showing
an antibacterial activity    M. Shibata, S. Koeda, R. Mitsuhashi, A. Miyagawa, H. Yamamura,T. Mizuno
Graduate School of Engineering, Nagoya Institute of Technology
Introduction
Bacterial infections cause various diseases, such as food poisoning, tuberculosis, andtetanus. At present, it is effective to use antibacterial reagents as a treatment for these
infections. However, due to an emergence of drug-resistant mutant bacteria, a development ofnovel antibacterial reagents is still necessary. Recently, we succeeded in evolving novel
solubilization surfactants for membrane proteins (DKDKC12K andDKDKC12D) from the PG-surfactant molecular framework (Figure1). Because there is a common mechanism to perturb bilayer
membranes on extracting membrane proteins from bilayer
membranes and lyzing bacterial bilayer membranes, development ofnovel antibacterial reagents from PG-surfactants was expected. In
this study, therefore, we explored novel antibacterial reagents from
PG-surfactants by a combinatorial screening from those having Figure 1. Chemical structureof PG-surfactant.
different amino acid sequences.
Results and Discussion
As a candidate derivatives, cationic or anionic PG-surfactants were synthesized by asolid-phase peptide synthesis. Antibacterial activity (minimum inhibitory concentration; MIC)
for E. coli and B. subtilis were evaluated by the broth microdilution method, and hemolytic
activity (the concentration at 50% hemolysis; HC50) for rabbit red blood cells was evaluated,
respectively. As results, the cationic and hydrophobic PG-surfactants showed a tendency to
exhibit higher antibacterial activities. But a higher hemolytic activity also oberved for thesame PG-surfactants.
Table. MIC and HC50 of PG-surfactants.

The number

K3C12
K4C12
K3C12’
K4C12’
K3C6
K4C6

X

Y

Carbonatoms

Lys-Lys-Lys
Lys-Lys-Lys-Lys
Lys-Lys-Lys
Lys-Lys-Lys-Lys
Lys-Lys-Lys
Lys-Lys-Lys-Lys

-

12
12
12
12
6
6

MIC (µg/ml)

Alkylchains E. coli
2
2
1
1
2
2

64
16
128
64
-

HC50

B.
subtilis

(µM)

8
8
8
4
128
64

8
3.5
135
85
> 600
> 600

73

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P34
  Direct Preparation of E-β-Ethynyl-α-stannylacrylate as Effective  
Building Block for Peridinin Synthesis by Novel
E-Selective Ethynylstannylation of Propiolate    
KINASHI, Naoto1; SAKAGUCHI, Kazuhiko1; SHINADA, Tetsuro1; KATSUMURA,
Shigeo2
1) Graduate School of Science, Osaka City University, Japan2) Osaka City University Advanced Research Institute for Natural Science & Technology(OCARINA)
Peridinin (1), a highly functionalized C37 norcarotenoid, has received much attention as alight-harvesting pigment in photosynthesis in the sea because of its quantitative energy
transfer efficiencies to chlorophyll a.[1] In order to elucidate the relationship between theunique chemical structure of peridinin and its super energy transfer ability, syntheses ofperidinin derivatives and their spectroscopic analyses have been studied.[2]In this poster , we would like to present a new synthetic approach to access peridinin (1)and its analogues using E-4 as a synthon for the ylidenbutenolide moiety. Stereoselective
synthesis of E-4 from 2 and 3 in one step, and an one-pot polyene elongation by crosscoupling reactions of E-4 will be present.

References:[1] Song, P. S.; Koba, P.; Prezelin, B. B.; Haxo, F. T. Biochemistry 1976, 15, 4422-4427.
[2] (a) Woerly, E. M.; Cherney, A. H.; Davis, E. K.; Burke, M. D. J. Am. Chem. Soc. 2010,
132, 6941–6943., (b) Furuichi, N.; Hara, H.; Osaki, T.; Mori, H.; Katsumura, S. Angew.
Chem., Int. Ed. 2002, 41, 1023-1026. (c) Kajikawa, T.; Aoki, K.; Singh, R. S.; Iwashita, T.;
Kusumoto, T.; Frank, H. A.; Hashimoto, H.; Katsumura, S. Org. Biomol. Chem. 2009, 7,
3723-3733.

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Structures and Properties ofBenzothiazinophenothiazine and its Cationic Species    
Kohei Yoshida, Shuichi Suzuki, Masatoshi Kozaki, Daisuke Shiomi,
Kazunobu Sato,Takeji Takui, and Keiji Okada
Department of Chemistry, Graduate School of Science, Osaka City University
Phenothiazine and its heteroatom analogues are highly electron-rich compounds becauseof their 8π electrons in the central six-membered rings. They have typically butterfly
structures in the neutral states, whereas their radical cation states have almost planar forms.
Benzothiazino-condensed phenothiazine and their oxygen analogue [1] are also interesting
because they have two oxidation sites. The derived dications could be open-shell singlet or
triplet, or closed singlet in the ground states. We here investigated the oxidation of
benzothiazinophenothiazine 1 in order to clarify the nature of the ground state.
Scheme 1. Oxidation reaction of 1.

The cyclic voltammogram of compound 1 indichloromethane showed two reversible oxidation
waves at −0.06 V (1 → 1•+) and +0.42 V (1•+ →12+) vs Fc/Fc+ (Figure 1). We also succeeded in theisolation of radical cation species 1•+ and dication
species 12+ by the chemical oxidation. Their
molecular structures in cationic states were
determined by X-ray crystal structure analyses. In
dicationic state, fused-ring moiety have planer

Figure 1. Cyclic voltammogram of 1

structures (Figure 2). Its C−S bonds and C−Nbonds are shorter than those of neutral state. Thesebonds shortening is rationalized by the possible
quinoidal structure of 12+.
[1] (a) Okamoto, T.; Kozaki, M.; Doe, M.; Uchida,M.; Wang, G.; Okada, K. Chem. Mater. 2005,
17, 5504–5511.

76

Figure 2. Ortep view of 12+.
Hydrogen atoms and counter anion
were omitted for clarity

P36

photo

In situ Huisgen cycloaddition reaction onto a surface of
membrane protein PSI using the novel alkynyl group-modified solubilization surfactants

KOEDA, Shuhei1; TANIGUCHI, Aki1; SHIBATA, Masahide1; NOJI, Tomoyasu2;
KAWAKAMI, Keisuke2; DEWA, Takehisa1; KAMIYA, Nobuo2; MIZUNO, Toshihisa1
1Grad. Sch. Eng., Nagoya Inst. of Tech.; 2OCARINA, Osaka City Univ.
Abstract: Recently, application ofvarious membrane proteins as a#" $" "$!##"
 "$!##
 !#!!%#
functional molecule is extensively%"" %"" %"studied. In order to utilize%"" %"""C
membrane proteins as a unit of
O O
Alk$C3$DKDKC12K+ %"" %""
nano devise, an immobilization
HC C
# $!##
in/onto suitable support materials
X=#$Asp$Lys$Asp$Lys$ "$!##
H 2N Cys X Cys Lys NH
should have several benefits. So far
Fig.1 Chemical structure of the series of PG-surfactants
we succeeded in constructing novel
solubilization surfactant for membrane proteins DKDKC12K that can be modified withS
S
various functional groups without losing original solubilization properties for membraneproteins[1]. In this research, we tested whether in situ Hüisgenreaction betweenO
O cycloaddition
alkynyl group, introduced in the solubilization surfactant
DKDKC12K, and azide
NH derivatives
HN
in a solution can be occurred without protein denaturation upon solubilizing membraneprotein Photosystem I (PSI) with the alkynyl group-modified surfactant(Alk-C3-DKDKC12K, Fig. 1).
Results and Discussion: The PSI solubilized with Alk-C3-DKDKC12K was prepared by asurfactant exchange method from the purified PSI


HN



S

S

HN Lys

NH

O

O

Cys

X

Cys

NH 2

solubilized with β-DDM. From the spectroscopic
properties (Fig. 2) and the light-induced electrontransfer activity, the PSI solubilized with
Alk-C3-DKDKC12K showed no denaturation. Upon
the addition of Azide-fluor 545, CuCl2, and sodium
ascorbate to this PSI solution, in situ Huisgencycloaddition reaction onto a PSI surface was
performed at 40 ˚C for 4 hr. From the SDS-PAGE
Fig.2 Absorption spectra of PSI, solubilized in aanalysis, we found quantitative Huisgen buffer containing 0.1 wt% β-DDM (red) and 0.1
cycloaddition reaction between Azide-fluor 545 wt% Alk-C3- DKDKC12K(blue).
and Alk-C3- DKDKC12K could be occurred without protein denaturation.
References: [1] S. Koeda & K. Umezaki & T. Mizuno, Langmuir 2013, 29, 11667.

77

CH

P37
Mesoporous material with photocatalyst towordhydrogen evolution
Tomoko Horibe1,2, Eiji Yamashita1,2, Hideki Hashimoto2 and Isamu Kinoshita2
1
Fuji Chemical Industry Co., Ltd., 1 Gohkakizawa Kamiichi-machi, Nakaniikawa-gun,Toyama 930-0397, Japan
2
Osaka City University Advanced Research Institute for Natural Science and Technology(OCARINA),3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, JapanFor the development of renewable energy and realize a low-carbon society, artificial
photosynthesis is important technology.
Neusilin is the fine powder of magnesium aluminometasilicate with amorphous structure
whose specific surface area is very large. Neusilin can carry various types of materials, thephotosynthetic membrane proteins, dyes, etc. Currently, the major use of the neusilin is in thepharmaceuticals. It is important to expand the utilization field, not limited thereto. Therefore,
we have been exploring the functional material to be introduced into Neusilin for the use of
artificial photosynthesis.
It is expected to stabilize the function of photocatalyst by combining a porous material.
This work target to build a light driven hydrogen production system using the photocatalyst in
combination with a porous material. For this purpose we have been developping
highly-efficient photocatalysts for the system.
In recent studies, the transition metal (Fe, Co, Ni) ppte and Me-ppte (Fig. 1 and 2)
complexes were prepared as the potential candidates of the photocatalyst. We eveluated theability for the hydrogen production of the complexes using a solution system containing
triethylamine as the sacrificial electron donor, fluorescein as the photosensitizer. Among ourcatalysts, complex CoCl2(Me-ppte)(Fig. 3) and [Ni (µ-Cl) (ppte)]2 (Fig. 4) achieve over 1400
and 3000 TON, respectively, in 30 h. We are confident that our catalyst will soon has come tothe point where soon catch up the highest TON =7300 reported among molecular noble
metal-free systems[1].
Cl

N
S

O
N

N

N

S

Fig.1 ppte

N

S

O
N

N

S

Fig. 2 Me-ppte

Cl
O

Co

N
S
S

N

N

N

Fig. 3 [CoCl2(Me-ppte)] S

S
O

C
C

N

Ni
N

Cl
Cl

N
Ni
N

C

C

S

O
S

Fig.4 [Ni(µ -Cl)(ppte)]2[1] Han, Z., Shen, L., Brennessel, W. W., Holland, P. L., Eisenberg, R.. J. Am. Chem. Soc.
2013, 135, 14659

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79