The90 th Anniversary MeetingThe

90 th Anniversary Meeting

The Society for Biotechnology, Japan
International Symposium on

Program & Abstracts

Program & Abstracts

October 24-26, 2012
Kobe International Conference Center

The Society for Biotechnology, Japan

Kobe, Japan

Organized by
The Society for Biotechnology, Japan (SBJ)
Japan Science and Technology Agency (JST)Supported by
Kobe Convention & Visitors Association
Nakauchi Tsutomu Convention Shinko Zaidan Foundation
Suntory Institute for Bioorganic Research

ERRATA◆The presentation below has been cancelled.
Abstract: Page 50

3Fa04 Production of the bio-butanol based on the continuous in situ

fermentation and adsorption (ISFA)

technology ႼHanjie YING1,2, Yong CHEN2,3, Jinglan WU2,3(1State Key Laboratory of Materials-Oriented Chemical
Engineering,China, 2 College of Biotechnology and Pharmaceutical Engineering,
China, 3 National Engineering

Nanjing University of Technology,

Technique Research Center for Biotechnology, China)

ご迷惑をおかけしましたことをお詫びし、ここに訂正いたします。
We apologize for any inconvenience this may have caused.

Table of Contents

Greetings................................................................................... 1
About SBJ ................................................................................. 2
Board Members ........................................................................ 3
Access ....................................................................................... 4
Floor Plan .................................................................................. 7
Program at a Glance ...............................................................11
Program ....................................................................................13
Abstracts ................................................................................. 27
Wednesday 24, October ............................................................ 29
Thursday 25, October.................................................................47
Friday 26, October .................................................................... 63
Index .........................................................................................81
Authors Index ........................................................................... 83
Keywords Index ........................................................................ 85

Greetings
Dear Colleagues,
We would like to welcome all of you to the 90th Anniversary Meeting of Society for
Biotechnology, Japan (SBJ), which will be held at the Kobe International
Conference Center (KICC), Kobe Port Island, from October 23rd (Tue) to 26th (Fri),
2012. To commemorate the 90th anniversary with all the participants, the meetingis organized with the concepts “from academia to industry”, “from Japan to theworld”, and “from senior to junior”.
At the opening day (October 23rd), the commemorative ceremony for the 90th
anniversary will be held in the morning, followed by award presentation ceremonyin the afternoon at the Main Hall of KICC. The celebration dinner will be held at
Kobe Portopia Hotel in the evening of October 23rd. From October 24th to 26th,
symposia, general sessions, and luncheon seminars will be held at the KICC
conference rooms, and technical exhibitions, including SBJ commemoration
displays, will be convened at the Reception Hall of KICC. There will also be theinternational symposium (all presentations and discussions are given in English)
on Biotechnology for Green Growth.
KICC and Kobe Portopia Hotel are both located in the Kobe Convention Complex
which is central to Port Island. Many restaurants, fashion towns, parks, andmuseums are scattered around Kobe, the city of convention, culture, and leisure.
The four-day long meeting will be an excellent opportunity for you to meet with
SBJ members and colleagues from abroad.
We are very pleased to welcome you to the exciting meeting which will be
remembered for years to come. Finally, we would like to thank all the members of
the Executive Committee, as well as our colleagues who contributed to thesuccess of this memorable event.
Hisao Ohtake
Chair of the Executive Committee
The 90th Anniversary Meeting

Satoshi Harashima
President
The Society for Biotechnology, Japan1

About SBJ

The Society for Biotechnology, Japan (SBJ), formerly the Society of Fermentation
Technology, Japan, was founded as the Osaka Brewing Society in 1923. SBJ
consists of approximately 3200 individual members, 250 institutional andsupporting members, representing throughout Asia as well as other regions.
SBJ monthly publishes journals in both English and Japanese, also provides
information and services online across the study of biotechnology. The Journal of
Bioscience and Bioengineering (JBB), first published as the Journal of
Fermentation Technology in 1973, is now highly appreciated by scientists
throughout the world (Impact factor in 2011: 1.793).
Events and seminars are conducted throughout a year. The annual meeting isheld in autumn covering for 3 days. Approximately 650 topics are discussed withmore than 1000 participants.
Acknowledging the world information is becoming more globalized each day. SBJ
aims to promote opportunities to network and exchange ideas among its members
to disseminate SBJ’s approaches; “the academia to the industry” and “tocontribute to the society” to the public.
Please visit our web site at http://www.sbj.or.jp/e/ for more information.

2

OrganaizaƟon CommiƩee

The 90th Anniversary Meeting of the Society for
Biotechnology, Japan
Board Members
President:
Vice President:
Chairperson:
Vice-Chairperson:

Satoshi Harashima
Kenji Sonomoto
Kenzo Yanagi
Hisao Ohtake
Eiichiro Fukusaki
Masahito TayaInternational Symposium Organizing Committee:
Kohsuke Honda
Shigeru Kitani
Ryo Misaki
Takuya Nihira
Hiroshi Umakoshi
Hitoshi Wake
International Symposium Secretariat:
The Society for Biotechnology, Japan
c/o Department of Biotechnology, Graduate School of Engineering,Osaka University
2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
Tel: +81-6-6876-2731, Fax: +81-6-6879-2034
E-mail: sympo2012@sbj.or.jp

3

Access

Kansai International Airport (KIX) is the closest international gateway for Kobe
and is located on the Osaka Bay.
Transfer from KIX to Kobe:
By Taxi: It takes about 70 min. and costs approx. 22,000yen (From KIX to
Sannomiya, downtown Kobe).
By Limousine Bus: It takes about 70 min. and costs 1,900yen (KIX to Sannomiya).
The bus runs every 10 to 20 min. from 6:20am till 10:40pm.

4

It takes about 2 hours and 50 min. and costs approx. 15,000yen from Tokyo toShin-Kobe by bullet train. The bullet train bound for Shin-Kobe from Tokyo
operates from 6am till 8:30pm. From Nagoya, it takes about 70 min. and costs
approx. 8,000yen. From Kyoto, it takes about 30 min. and costs approx. 3,500yen.
Transfer from Shin-Kobe to downtown Kobe, Sannomiya:
By Taxi: It takes about 5 min. and costs approx. 660yen.
By City Subway: It takes about 2 min. and costs 200yen.

It takes about 20 min. and costs approx. 400yen by JR Lines from Osaka.
It takes about 50 min. and costs approx. 1,050yen from Kyoto.

“Port Liner” monorail: The monorail bound for Kobe Airport from Sannomiya
station runs every 5 min. and stops at “Shimin Hiroba”, the station closest to the
Kobe International Coference Center. From Sannomiya station to Shimin Hiroba
station, it takes about 10 min. and costs 240yen by Port Liner.
By Taxi: It takes about 10 min. and approx. 1500yen from Sannomiya.

5

ACCESS

By Bay Shuttle Ferry (to Kobe Airport): It takes about 30 min. and costs 1,800yen.
The ferry runs every 45 to 60 min. from 7:15am till 10:00pm. Free bus transfer isavailable from KIX to KIX Ferry dockland. From Kobe airport, take Port Linermonorail to Shimin Hiroba, the station closest to the Kobe International
Conference Center.

6Floor Plan

Floor Plan

7

Kobe International Conference Center BF

Kobe International Conference Center 1F

8

Floor Plan

Kobe International Conference Center 3F

Kobe International Conference Center 4F

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11

Floor Plan

Program at a GlanceProgramWed 24, OCT

10/24 (Wednesday)
Advances in PhytoGenomics
Organizers: Toshiya MURANAKA (Osaka University)
Kiichi FUKUI (Osaka University)
Chairperson: Kiichi FUKUI (Osaka University)
9:00–
9:25

2Fa01

Functional chromosome dynamics with a link to synthetic
green biotechnology○Ingo SCHUBERT

9:25–
9:45

2Fa02

Manipulation of plant traits using novel gene silencing system,
CRES-T (Chimeric REpressor gene Silencing Technology)○Nobutaka MITSUDA1, Masaru OHME-TAKAGI1,2(1Bioproduction Research Institute, National Institute of Advanced
Industrial Science and Technology (AIST), Japan, 2Institute forEnvironmental Science and Technology (IEST), Saitama University,
Japan)

9:45–
10:05

2Fa03

A top-down approach to function the Synechocystis PCC6803
genome in a novel cloning host Bacillus subtilis 168○Mitsuhiro ITAYA(Institute for Advanced Biosciences, Keio University, Japan)

Chairperson: Toshiya MURANAKA (Osaka University)
10:05–
10:30

2Fa04

Decoding the chemical diversity and evolution written in the
genomes of the Asteraceae○Dae-Kyun RO1, Don Trinh NGUYEN1, Nobuhiro IKEZAWA 2,
Vince QU1
(1Department of Biological Sciences, University of Calgary, Alberta,
Canada, 2Department of Biotechnology, Osaka University, Japan)

10:30–
10:55

2Fa05

Cassava molecular breeding and functional genomics–Challenges and opportunities–○Jarunya NARANGAJAVANA(Center for Cassava Molecular Biotechnology, Department of
Biotechnology, Faculty of Science, Mahidol University, Bangkok,
Thailand)

10:55–
11:15

2Fa06

KOMICS: metabolomics databases and tools for biotechnology○Daisuke SHIBATA, Takeshi ARA, Hideyuki SUZUKI,
Nozomu SAKURAI(Kazusa DNA Research Institute, Chiba, Japan)

15

Program(Department of Cytogenetics and Genome Analysis, Leibniz
Institute of Plant Genetics and Crop Plant Research (IPK), Germany)

Wed 24, OCT

Chairperson: Kazuhito FUJIYAMA (Osaka University)
11:15–
11:40

2Fa07

Dammarene-type ginsenoside metabolic engineering–Characterization of genes involved in dammarenediol saponinbiosynthesis–
Jung-Yeon HAN, ○Yong-Eui CHOI
(Department of Forest Resources, College of Forest andEnvironmental Science, Kangwon National University, South Korea)

11:40–
12:00

2Fa08

Production of plant triterpenoids in engineered yeast○Toshiya MURANAKA, Hikaru SEKI

Program(Department of Biotechnology, Graduate School of Engineering,Osaka University, Japan)

Bio-Based Production
Organizers: Hiroshi SHIMIZU (Osaka Univeristy)
Eiichiro FUKUSAKI (Osaka University)
Chairperson: Hiroshi SHIMIZU (Osaka Univeristy)
13:50–
14:20

2Ap01 Living microbes as selective and efficient redox biocatalysts○Andreas SCHMID, Katja BÜHLER, Mattijs JULSING,
Bruno BÜHLER
(Laboratory of Chemical Biotechnology, TU Dortmund University,
Germany)

14:20–
14:50

2Ap02 Toward biological replacement of petroleum○James C. LIAO
(Department of Chemical and Biomolecular Engineering,
University of California, Los Angeles, USA)

14:50–
15:20

2Ap03 Bacillus subtilis cell factory for production of scyllo-inositol
promising for Alzheimer’s disease○Ken-ichi YOSHIDA1, Chumsakul ONUMA 2, Shu ISHIKAWA 2,
Naotake OGASAWARA 2(1 Department of Agrobioscience, Graduate School of Agricultural
Science, Kobe University, Japan, 2 Graduate School of Biological
Sciences, Nara Institute of Science and Technology (NAIST), Japan)

15:20–
15:50

2Ap04 Engineering of signal transduction for enhanced production ofbiopharmaceuticals and bioenergy○Jian-Jiang ZHONG1, Yang-Chun YONG2(1 School of Life Sciences and Biotechnology, Shanghai Jiao Tong
University, China, 2 School of the Environment, Jiangsu University,
China)

16

2Ap05 Synthetic biology for the comprehension of biomolecular
networks○Masahiro OKAMOTO1,2(1 Department of Bioinformatics, Graduate School of Systems Life
Sciences, Kyushu University, Japan, 2 Synthetic Systems Biology
Research Center (SSBRC), Kyushu University, Japan)

16:20–
16:30

BREAK

2Ap06 Strategies and technologies towards tailor made fuels frombiomass○Jochen BUECHS1, Tobias KLEMENT1, Martin KUNZE1,
Gernot JAEGER1, Sandra WEWETZER1,Frederike CARSTENSEN2, Matthias WESSLING2,
Philipp GRANDE3, Pablo Domínguez DE MARIA3, Antje SPIESS4(1 AVT - Biochemical Engineering, RWTH Aachen University,
Germany, 2 AVT - Chemical Process Engineering, RWTH Aachen
University, Germany, 3 Institute of Technical and Macromolecular
Chemistry, RWTH Aachen University, Germany, 4 AVT - Enzyme
Process Technology, RWTH Aachen University, Germany)

17:00–
17:30

2Ap07 Actinobacteria tyrosinase and its applications to biomaterials○Byung-Gee KIM1,2,3, Nahum LEE1,3, SangHyuk LEE2,3,
ChangHyun SONG1,3(1 School of Chemical and Biological Engineering, Seoul National
University, Korea, 2 Institute of Molecular Biology and Genetics,
Seoul National University, Korea, 3 Institute of Bioengineering,Seoul National University, Korea)

17:30–
18:00

2Ap08 Cyberinfrastructure for metabolomics and synthetic biology○Masanori ARITA1,2(1Department of Biophysics and Biochemistry, Graduate School of
Science, The University of Tokyo, Japan, 2 RIKEN Plant Science
Center, Japan)

18:00–
18:30

2Ap09 Bioprocess engineering on biofuels and bio-based chemicals
production from microalgae○Jo-Shu CHANG1,2,3, Chun-Yen CHEN2(1Department of Chemical Engineering, National Cheng Kung
University, Taiwan, 2Center for Bioscience and Biotechnology,
National Cheng Kung University, Taiwan, 3Research Center for
Energy Technology and Strategy, National Cheng Kung University,
Taiwan)17

ProgramChairperson: Eiichiro FUKUSAKI (Osaka University)
16:30–
17:00

Wed 24, OCT

15:50–
16:20

Wed 24, OCT

18:30–
19:00

2Ap10 Systems metabolic engineering –Rational design of microbial
cell factories○Hiroshi SHIMIZU1, Chikara FURUSAWA1,2, Takashi
HIRASAWA1, Naoaki ONO3, Katsunori YOSHIKAWA1,
Yoshihiro TOYA1

Program(1 Department of Bioinformatic Engineering, Osaka University,
Japan, 2 Quantitative Biology Center (QBiC), RIKEN, Japan,
3
Graduate School of Information Science, Nara Institute of
Science and Technology (NAIST), Japan)

18Thu 25, OCT

10/25 (Thursday)
Biomolecular Engineering [co-organized by Young Asian
Biochemical Engineers’ Community (YABEC)]
Organizers: Takeshi OMASA (University of Tokushima)
Noriho KAMIYA (Kyushu University)
Chairperson: Takeshi OMASA (University of Tokushima)
9:00–
9:30

3Fa01

Reaction evaluation and new process design in composting of
biological wastes~Young Asian Biotechnologist Prize~○Jingchun TANG1, Arata KATAYAMA 2

9:30–
9:40

BREAK

Chairperson: Noriho KAMIYA (Kyushu University)
9:40–
10:05

3Fa02

Organic solvent stable enzymes○Hiroyasu OGINO(Department of Chemical Engineering, Osaka Prefecture University,
Japan)

10:05–
10:30

3Fa03

Engineering of aminoacyl-tRNA synthetases for residuespecific incorporation of amino acid analogues into proteins○Tae Hyeon YOO(Department of Molecular Science and Technology, Department ofApplied Chemistry and Biological Engineering, Ajou University,
Korea)

10:30–
10:55

3Fa04

Production of the bio-butanol based on the continuous in situ
fermentation and adsorption (ISFA) technology○Hanjie YING1,2, Yong CHEN2,3, Jinglan WU2,3(1State Key Laboratory of Materials-Oriented Chemical Engineering,
China, 2 College of Biotechnology and Pharmaceutical Engineering,
Nanjing University of Technology, China, 3 National Engineering
Technique Research Center for Biotechnology, China)

19

Program(1College of Environmental Science and Engineering, Nankai
University, China, 2 EcoTopia Science Institute, Nagoya University,
Japan)

Thu 25, OCT

Chairperson: Takeshi OMASA (University of Tokushima)
10:55–
11:20

3Fa05

Functional properties of diet ginger (Zingiber officinale Roscoe,
Zingiberaceae) for antioxidant, anti-pigmentation, anti-cancer,
anti-bacterial and wound repair
Pei-Fang WU, Yi-Ting CHOU, ○Hui-Min WANG
(Department of Fragrance and Cosmetic Science, Kaohsiung
Medical University, Kaohsiung, Taiwan, ROC)

11:20–
11:45

3Fa06

Enzyme engineering with the aid of ionic liquids○Noriho KAMIYA1,2

Program(1Center for Future Chemistry, Kyushu Univeristy, Japan,
2
Department of Applied Chemistry, Graduate School ofEngineering, Kyushu University, Japan)

SBJ/JST Joint Symposium I –General Session–
Organizers: Tadashi MATSUNAGA (Tokyo University of Agriculture and Technology)
Hisao OHTAKE (Osaka University)
Shigeru ISHIMASA (JST)
Chairperson: Kohsuke HONDA (Osaka University)
14:00–
14:05

Welcome adress○Hisao OHTAKE(Department of Biotechnology, Graduate School of Engineering,Osaka University, Japan)

14:05–
14:15

3Ap01 Introduction of the JST CREST/PRESTO program researcharea “Creation of basic technology for improved bioenergy
production through functional analysis and regulation of algae
and other aquatic microorganisms”○Tadashi MATSUNAGA(Tokyo University of Agriculture and Technology, Japan)

Chairperson: Masafumi YOHDA (Tokyo University of Agriculture and Technology)
14:15–
14:50

3Ap02 Can biotechnology address the global energy crisis?
○Jonathan D. TRENT1,2(1 Bioengieering Branch, NASA Ames Research Center, USA,
2
Biomolecular Engineering, University of California Santa Cruz, USA)20

14:50–
15:25

3Ap03 Food and fuel from microalgae: Integration of photosynthesis
and photovoltaics allows to achieve a positive energy balanceof algae biomass production○Mario R. TREDICI

Thu 25, OCT

Chairperson: Akira SATO (Yamaha Motor, Co.)

(University of Florence, Italy)
15:25–
15:35

BREAK

15:35–
16:10

3Ap04 Biorefinery of microalgae○René H. WIJFFELS(Wageningen University, Bioprocess Engineering, the Netherlands)

Chairperson: Kohsuke HONDA (Osaka University)
16:10–
16:20

Closing remarks○Shigeru ISHIMASA(Japan Science and Technology Angency (JST), Japan)

Japan-Korea Biomass Session
Organizers: Atsuhiko SHINMYO (Nara Institute of Science and Technology)
Shigeaki FUJIKAWA (NPO Kinki Bio-Industry Development Organization)
Chairperson: Shigeaki FUJIKAWA (NPO Kinki Bio-Industry Development Organization)
16:40–
16:45

Opening remarks○Atsuhiko SHINMYO(Nara Institute of Science and Technology, Japan)

16:45–
17:05

3Ap05 Improvement of ethanol yield from xylose by breeding of industrial
yeast○Haruyo HATANAKA, Tomohide MAEDA, Yuri YASUE, KazueNISHIYAMA(Suntory Business Expert Limited, Institute for Microbial Science,
Japan)

21

ProgramChairperson: Tsuyoshi TANAKA (Tokyo University of Agriculture and Technology)

Thu 25, OCT

17:05–
17:25

3Ap06 Applications of microbial factory technology to production ofbioethanol from cellulosic biomass
Jung-Hyun JO1, Sang-Min JUNG1, Yong-Cheol PARK 2,
Yong-Su JIN3, ○Jin-Ho SEO1

Program(1Department of Agricultural Biotechnology, Seoul National
University, Korea, 2 Department of Advanced Fermentation Fusion
Science and Technology, Kookmin University, Korea, 3 Department
of Food Science and Human Nutrition, University of Illinois at
Urbana-Champaign, USA)
17:25–
17:45

3Ap07 Potential in bioethanol production from waste paper sludge inpulp-based biorefinery○Enoch Y. PARK1,2, Joni PRASETYO1, Kazuya NARUSE2,
Tatsuya KATO2(1 Laboratory of Biotechnology, Graduate School of Science andTechnology, Shizuoka University, Japan, 2 Laboratory of
Biotechnology, Faculty of Agriculture, Shizuoka University, Japan)

17:45–
17:55

BREAK

Chairperson: Moon-Hee SUNG (Kookmin University)
17:55–
18:15

3Ap08 Application of Metabolomics to high resolution phenotype
analysis○Eiichiro FUKUSAKI(Department of Biotechnology, Graduate School of Engineering,Osaka University, Japan)

18:15–
18:35

3Ap09 Biological production of 3-hydroxypropionic acid from glycerol○Sunghoon PARK
(School of Chemical and Biomolecular Engineering, Pusan National
University, Korea)

18:35–
18:55

3Ap10

Biodiesel production by enzymatic process and the utilizationof crude glycerol
Hah Young YOO, Laxmi Prasad THAPA, Ja Hyun LEE,
Sang Jun LEE, ○Seung Wook KIM(Department of Chemical and Biological Engineering, Korea
University, Korea)

18:55–
19:00

Closing remarks○Sunghoon PARK
(Pusan National University, Korea)

22

Fri 26, OCT

10/26 (Friday)
SBJ/JST Joint Symposium II –Biorefinery–
Organizers: Haruko TAKEYAMA (Waseda University)
Yoshihiro SHIRAIWA (Tsukuba University)
Chairperson: Haruko TAKEYAMA (Waseda University)
9:00–
9:25

4Aa01

Understanding and altering the unique metabolism ofhyperthermophilic archaea○Haruyuki ATOMI1,2, Kunio MIKI2,3, Tadayuki IMANAKA 2,4

9:25–
9:50

Program(1Department of Synthetic Chemistry and Biological Chemistry,
Graduate School of Engineering, Kyoto University, Japan,
2
CREST, Japan Science and Technology Agency (JST),
3
Department of Chemistry, Graduate School of Science, Kyoto
University, Japan, 4Department of Biotechnology, College of Life
Sciences, Ritsumeikan University, Japan)

4Aa02 Blue ocean for blue biotechnology – Marine bioenergy
production○Choul-Gyun LEE1,2(1Department of Biotechnology, Inha University, Korea,
2
Korea National Consortium on Marine Bioenergy Research)

9:50–
10:15

4Aa03 Establishment of innovative technology to create new
microalgal strains increasing biofuel production by
polyploidization and heavy-ion beam irradiation○Shigeyuki KAWANO1,2, Koichi WATANABE1,2, Shuhei OTA1,2(1Department of Integrated Biosciences, Graduate School of Frontier
Sciences, The University of Tokyo, Japan, 2CREST, Japan Scienceand Technology Agency (JST), Japan)

Chairperson: Yoshihiro SHIRAIWA (Tsukuba University)
10:15–
10:40

4Aa04 New dimensions in microalgae biomass production forbioenergy○Otto PULZ
(IGV, Nuthetal, Germany)

10:40–
10:45

BREAK

23

Fri 26, OCT

Chairperson: Yoshihiro SHIRAIWA (Tsukuba University)
10:45–
11:10

4Aa05 Development of sustainable biorefinery based on microalgae○Akihiko KONDO1,2(1Department of Chemical Science and Engineering, Graduate
School of Engineering, Kobe University, Japan, 2Core Research
for Evolutional Science and Technology (CREST), Japan Scienceand Technology Agency (JST), Japan)

Program

11:10–
11:35

4Aa06 Chlamydomonas reinhardtii - A powerful host for the productionof biofuels and high value products○Olaf KRUSE(Bielefeld University, Department of Biology/ Center for Biotechnology,
Algae Biotechnology & Bioenergy Group, Germany)

Chairperson: Haruko TAKEYAMA (Waseda University)
11:35–
12:00

4Aa07 Alkenone biosynthesis by marine Haptophytes as biorefinery
for renewable energy production○Yoshihiro SHIRAIWA1,2(1Faculty of Life & Environmental Sciences, University of Tsukuba,
Japan, 2 CREST, Japan Science and Technology Agency (JST),Japan)

SBJ/JST Joint Symposium III –Biofuel Production by Microalgae–
Organizers: Masayuki OHMORI (Chuo University)
Shigeru OKADA (The University of Tokyo)
Chairperson: Masayuki OHMORI (Chuo University)
14:00–
14:20

4Ap01 Characterization of hydrocarbon biosynthesis and secretionmechanisms by the green microalga, Botryococcus braunii tocontrol biofuel production○Shigeru OKADA1,5, Ikuro ABE2,5, Tetsuko NOGUCHI3,5,
Takeshi OHAMA4,5(1Department of Aquatic Biosciences, Graduate School of Agriculturaland Life Sciences, The University of Tokyo, Japan, 2 Graduate Schoolof Pharmaceutical Sciences, The University of Tokyo, Japan,
3
Department of Biological Sciences, Faculty of Science, Nara
Women’s University, Japan, 4 Department of Environmental Systems
Engineering, Kochi University of Technology, Japan, 5 CREST, Japan
Science and Technology Agency (JST), Japan)

24

Fri 26, OCT

14:20–
14:55

4Ap02 Metabolic engineering of cyanobacteria for the light-powered
production of hydrogen from water○Matthias RÖGNER
(Plant Biochemistry, Faculty of Biology & Biotechnology, RuhrUniversity Bochum, Germany)

14:55–
15:15

4Ap03 Selection strategies for the promising microalgal strains foralgae-fuel production○Hideaki MIYASHITA1, Norihide KURANO2

15:15–
15:25

BREAK

Chairperson: Shigeru OKADA (The University of Tokyo)
15:25–
16:00

4Ap04 Elucidation of novel triterpene pathways in Botryococcus andengineering plants for high-value oil production
Tom D. NIEHAUS1, Scott KINISON1, Shigeru OKADA 2,
Shuiqin WU1, Timothy P. DEVARENNE3, David S. WATT4,
Vitaliy SVIRIPA4, ○Joe CHAPPELL1(1Plant Biology Program, University of Kentucky, USA, 2Department
of Aquatic Biosciences, University of Tokyo, Japan, 3Department ofBiochemistry and Biophysics, Texas A&M University, USA,
4
Department of Cellular and Molecular Biochemistry, University of
Kentucky, USA)

16:00–
16:20

4Ap05 The CyanofactoryTM~ a novel cyanobacterial bioprocess basedon the synthetic biology concept ~○Koji SODE1,2(1Tokyo University of Agriculture & Technology, Japan, 2 Japan
Science and Technology Agency, CREST, Japan)

25

Program(1Department of Interdisciplinary Environment, Graduate School of
Human and Environmental Studies, Kyoto University, Japan,
2
Research laboratories, DENSO CORPORATION, Japan)

Fri 26, OCT

SBJ/JST Joint Symposium IV –System & Synthetic Biotechnology–
Organizers: Daisuke UMENO (Chiba University)
Tomohisa HASUNUMA (Kobe University)
Kohsuke HONDA (Osaka University)
Chairperson: Daisuke UMENO (Chiba University)

Program

16:40–
17:00

4Ap06 Bio-refinery from microalgae through fermentation andmetabolic engineering○Tomohisa HASUNUMA1,2(1Organization of Advanced Science and Technology, Kobe
University, Japan, 2PRESTO, Japan Science and Technology
Agency (JST), Japan)

17:00–
17:35

4Ap07 Synthetic biology approaches to produce C3-C5 alcohols frommicroorganisms○Shota ATSUMI(Department of Chemistry, University of California Davis, USA)

17:35–
17:55

4Ap08 Synthetic metabolic engineering –A novel, simple technology
for designing a chimeric metabolic pathway–○Kohsuke HONDA1,2, Xiaoting YE1, Kenji OKANO1,
Hisao OHTAKE1(1 Department of Biotechnology, Graduate School of Engineering,Osaka University, Japan, 2 PRESTO, Japan Science and Technology
Agency (JST), Japan)

17:55–
18:05

BREAK

Chairperson: Tomohisa HASUNUMA (Kobe University)
18:05–
18:40

4Ap09 Production of BiolsopreneTM monomer○Derek H. WELLS(Du Pont Industrial Biosciences, USA)

18:40–
19:00

4Ap10 Evolutionary pathway engineering: toward the fast-track
construction of long-step, but high-fidelity pathway for nonnatural compounds○Daisuke UMENO1,2, Maiko FURUBAYASHI1, Kyoichi SAITO1(1Department of Applied Chemistry and Biotechnology, Graduate
School of Engineering, Chiba University, Japan, 2PRESTO, Japan
Science and Technology Agency (JST), Japan)

26

Abstracts

Advances in PhytoGenomics

Functional chromosome dynamics with a link to synthetic green
biotechnology

Wed 24, OCT

2Fa01○Ingo SCHUBERT
Department of Cytogenetics and Genome Analysis, Leibniz Institute of Plant Genetics
and Crop Plant Research (IPK), D-06466 Gatersleben, Germany[Keywords] Chromosome biology, Interphase chromatin dynamics, Centromere
maintenance, Artificial chromosomes, Synthetic centromeres

29

Abstracts

One of the research focuses of the IPK in Gatersleben is dynamics of plant
genomes. In the frame of this topic, groups of the Department of Cytogenetics
and Genome Analysis work on several aspects of chromosome biology applying
interphase cytogenetics, chromosome painting and other cutting-edge technologies.
Aims are to elucidate i) karyotype evolution, e.g. how large or how small can achromosome be, which mechanisms shape the dipoid chromosome complement,
how to trace the chromosomal ‘diploidisation’ of (allo)polyploids (mesopolyploidy),
but also ii) structural arrangement of chromosomes and their distinct functional
domains (centromeres, telomeres, nucleolus organizers, eu- and heterochromatin)
during interphase and during nuclear divisions, iii) correlations of specificepigenetic modifications with competent versus incompetent functional states; iv)
degree and functional importance of dynamics of interphase nuclear architecture.
I will provide a glancing light on some selected examples of this research andeventually on aspects of potential applicability, such as the generation of artificial(mini)chromosomes for breeding and biotech purposes.

Wed 24, OCT

2Fa02

Advances in PhytoGenomics

Manipulation of plant traits using novel gene silencing system,
CRES-T (Chimeric REpressor gene Silencing Technology)○Nobutaka MITSUDA1, Masaru OHME-TAKAGI1,2
1

Abstracts

Bioproduction Research Institute, National Institute of Advanced Industrial Scienceand Technology (AIST)
2
Institute for Environmental Science and Technology (IEST), Saitama University

A number of transcription factors have been shown to play important roles for the
regulation of growth and development and act as master regulator of various
phenotypes in plants. Thus, identification of biological function of eachtranscription factor is an important subject for elucidation of transcriptional
network, signaling cascade and manipulation of plant traits. However, plant genes
are frequently duplicated and this structural and functional redundancy often
interferes with efforts to identify the functions of these factors. To overcome these
difficulties, we developed a novel gene silencing system, called Chimeric
REpressor Gene-Silencing Technology (CRES-T), in which a transcription factor
that is converted into strong repressor by fusion with EAR-motif repression
domain (SRDX) suppresses the expression of target genes dominantly over theactivation activity of endogenous and functionally redundant transcription factors,
resulted in the induction of phenotype similar to loss-of-function of the allelesof the gene for transcription factor. Using this CRES-T system, we arecomprehensively analyzing function of transcription factors encoded in
Arabidopsis genome. In addition, we isolated a number of CRES-T lines thatexhibit tolerance to various abiotic stresses, such as high salinity, high osmoticpressure, heat, freezing and drought. Not only in model plants, we found that thechimeric repressor is a useful to manipulate various plant traits and actually
produced horticultural plants with different shape of flowers. We present that the
CRES-T system is not only useful for functional analysis of redundant transcription
factors in plants but also is a powerful tool for the manipulation of plant traits.
[Keywords] Transcription factor, Chimeric repressor, Gene regulation30

Advances in PhytoGenomics

A top-down approach to function the Synechocystis PCC6803
genome in a novel cloning host Bacillus subtilis 168

Wed 24, OCT

2Fa03○Mitsuhiro ITAYA
Institute for Advanced Biosciences, Keio University[1] Itaya, et al., (2005) PNAS, 102, 15971.
[2] Gibson, et al., (2010) Science, 329, 52[3] Itaya, (2009) p155. In Systems Biology and Synthetic Biology, Pengcheng etal., Eds, John Wiley & Brothers, Inc.
[Keywords] Genome design, Whole genome cloning, Gene expression, RibosomalRNA

31

Abstracts

Gene networks of exiting genomes are well regulated to make cellular lives invarious environments. Genome engineering recently achieved enables top down
approach to examine all the genes included in one genome species albeit limited
to small bacteria [1, 2]. Highlighted was a demonstration of chimera genome
comprised with a 3.5-Mb genome of a nonpathogenic, unicellular photosynthetic
bacterium Synechocystis PCC6803 and a 4.2-Mb genome of Bacillus subtilis 168[1].
The chimera genome cell, stemmed from B. subtilis to which the entire
cyanobacterium genome was assembled, has not grown yet in photosynthesis
media. Most of genes from the cyanobacterium genome were dormant and theirexpression mode was found unregulated in both transcription and translation andmolecular regulation mechanism is essential to produce cells carrying designed
genomes [3]. Certain biological significance raised by the newly constructedchimera genome and our attempts to draw scenarios for conversion of the two
network systems existing in one cell will be presented.
In contrast to E. coli host-vector systems, giant DNA equivalent to bacterial
genome are being effectively handled in our system using the Bacillus GenoMe(BGM) vector derived from the genome of B. subtilis 168 [1, 3]. Methods to handle
other giant DNAs form plants and animals and their plausible applications would
be shown.

Wed 24, OCT

2Fa04

Advances in PhytoGenomics

Decoding the chemical diversity and evolution written in the
genomes of the Asteraceae○Dae-Kyun RO1, Don Trinh NGUYEN1, Nobuhiro IKEZAWA 2, Vince QU1
1

Department of Biological Sciences, University of Calgary, Alberta, Canada
Department of Biotechnology, Osaka University, Japan

Abstracts

2

A recent development in next generation sequencings (NGS) broke the boundary
between “model” and “non-model” organisms and accordingly has provided
unique opportunities to elucidate the biosynthetic mechanisms of many untapped
phytochemicals in medicinal plants. Sunflower (or Asteraceae) family is one of theunder-studied plant families, despite that it constitutes ~8% of all angiosperms(~24,000 species). Asteraceae has evolved from the South America at about 50
million years ago and has rapidly radiated to other continents to become adominant plant family on earth. Among many specialized metabolites, two mainchemical constituents in Asteraceae are sesquiterpene lactone (STL) and natural
rubber (NR). However, their biosynthesis and regulation remain unknown. We
conjectured that the studies of STL and NR metabolism in 50 million years oftime-frame will help us better understand how specialized metabolisms evolveand diversify in plant. Using the NGS data, the genes coding for the central
oxidizing enzyme in the STL biosynthesis were identified from several subfamilies of Asteraceae, including a fossil plant, Barnadesia spinosa and a recently
evolved species, Artemisia annua. Comparative studies of their enzymatic
activities have shown that the primordial enzyme from B. spinosa displays abroader range of substrate specificity than recently emerged enzymes. Novelterpene compounds could be synthesized by pairing the primordial enzyme withvarious sesquiterpene substrates. Genomics analysis in lettuce further identified
a cytochrome P450 which catalyzes the formation of the simplest STL,
costunolide, but its homolog in sunflower catalyzes distinct regio- andstereoselective hydroxylation on the same sesquiterpene substrate. Homology
structural modeling of these two P450 enzymes revealed a small number ofresidues differing in their substrate-binding sites. However, simple exchange of
those residues resulted in the loss of activities but did not swap their activities.
This result implies that an additional layer of residues contribute to the specificity.
In addition to STL, lettuce and sunflower can synthesize terpene-polymer NR withdifferent polymer lengths. Genomics and reverse genetics approach in lettuce toelucidate NR biosynthesis will be also discussed.
[Keywords] Next generation
Cytochrome P450, Asteraceae

32

sequencing,

Sesquiterpene,

Naturalrubber,Advances in PhytoGenomics

Cassava molecular breeding and functional genomics–Challenges and opportunities–

Wed 24, OCT

2Fa05○Jarunya NARANGAJAVANA
Center for Cassava Molecular Biotechnology, Department of Biotechnology,
Faculty of Science, Mahidol University, Bangkok, Thailand[Keywords] Cassava, Energy crop, Genome, Molecular breeding, Functional
genomics

33

Abstracts

Cassava (Manihot esculenta Crantz) is the third most important source of calories
in the tropics, after rice and maize. The high starch content makes cassava adesirable energy source for human consumption as well as for various industrial
bio-products including biofuel applications. Cassava breeding efforts throughout
the world have made significant impact on cassava production, however it isanticipated that additional gains in breeding efficiency, which would translate into
genetic gain, could be made through the application of advanced molecular
breeding technologies. The cassava whole genome sequence, recently wasdeveloped by the US-DOE Joint Genome Institute Community Sequencing
Program, facilitates the implementation of molecular tools. It allowed theidentification of candidate simple sequence repeat and single nucleotide
polymorphism markers which can be used for genetic mapping and marker
assisted selection for further improvement. The availability of a well-annotated,
full-length cassava reference genome has been instrumental in facilitating
valuable transcriptome, proteome, and functional genomics applications incassava. We focus on identifying genes and quantitative trait loci controlling
economically important traits. Better understanding of how genes play suchimportant roles involved in several traits has been investigated. It is critical forpolicy-makers, the research community, and donors to understand the challenges
and opportunities of developing this crop for the future. The regional cooperation
should help in clearly defining the target problems and challenges to rise up to
the expectations of stake holders.

Wed 24, OCT

2Fa06

Advances in PhytoGenomics

KOMICS: metabolomics databases and tools for biotechnology○Daisuke SHIBATA, Takeshi ARA, Hideyuki SUZUKI, Nozomu SAKURAI

Abstracts

Kazusa DNA Research Institute, Chiba, Japan

Metabolomics approaches have been prevailing in biotechnology research during
the past decade. However, archiving information of metabolites found in variousorganisms and integrating the metabolome data to the transcriptome data of eachorganism are still a challenging field in bioinformatics. With recent advance intechnologies of mass spectrometry (MS), low molecular weight metabolites areanalyzed both in a high resolution and sensitivity, which leads to reliable
quantification of each metabolite, with help of advanced software for handlingchromatogram data, which is prerequisite for archiving metabolite data as
metabolomics research. Due to the large chemical diversity found in metabolites,
various MS machines with state-of-the-art technologies have prevailed formetabolite analysis. Over the past decade our laboratory has been analyzing
metabolites from various organisms and authentic chemicals for metabolite
identification using eight MS machines and accumulated more than 100,000
chromatograms in our servers. With intension to provide the whole datasets topublic in a unified manner to accelerate the metabolomics science, we havedesigned archives for data depository, as the databases MassBase for MS
chromatogram datasets and KomicMarket for annotated metabolite datasets.
Furthermore, we have been devoting to the development of genetic information
databases and tools. One of the databases, the KaPPA-View systems are foroverviewing the metabolome and transcriptome data on the pathway maps, as an
option, with the gene-to-gene or metabolite-to-metabolite co-relationships. These
databases and tools are available through our web portal KOMICS (Kazusa
Metabolomics Database) (http://www.kazusa.or.jp/komics/en/).
[Keywords] Metabolome
spectrometry

34

analysis,

Transcriptome,

Bioinformatics,

Mass

Advances in PhytoGenomics

Dammarene-type ginsenoside metabolic engineering–Characterization of genes involved in dammarenediol saponinbiosynthesis–

Wed 24, OCT

2Fa07

Jung-Yeon HAN, ○Yong-Eui CHOI
Department of Forest Resources, College of Forest and Environmental Science,
Kangwon National University, Chunchon 200-701, South Korea[Keywords] Panax ginseng,Dammarane-type ginsenoside

Tetracyclic

triterpene,

Saponinbiosynthesis,

35

Abstracts

Ginseng (Panax ginseng C.A. Meyer) is one of the most popular medicinal herbs
and contains pharmacologically active components, ginsenosides, in their roots.
Ginsenosides, a class of tetracyclic triterpene saponins, are synthesized fromdammarenediol-II after hydroxylation by the cytochrome P450 (CYP) enzymes
and then glycosylation by glycosyltransferases. The first step in biosynthesis ofdammarane-type ginsenosides is cycling of 2, 3-oxidosqualene to dammarenediol-II,a reaction that is catalyzed by an enzyme from dammarenediol synthase. We
functionally characterized the dammarenediol synthases (PgDDS) from P. ginseng.
Dammarenediol-II is thought to be converted to two ginsenoside aglycones(protopanaxadiol and protopanaxatriol) after hydroxylation by cytochrome P450
(CYP) enzymes. Two CYP genes are thought to be involved in dammarene-typeginsenoside biosynthesis. One of these genes is involved in protopanaxadiol
production by hydroxylation of dammarenediol at the C-12 position. Another geneis protopanaxatriol synthase which involved in protopanaxadiol hydroxylation at
the C-6 position conferring protopanaxatriol production. We reported that
CYP716A47 is involved in the hydroxylation of dammarenediol-II at the C-12
position to yield protopanaxadiol (Han et al. 2011). The next step of protopanaxatriol
production (protopanaxdiol 6-hydroxylase) is also characterized by our group
recently. We will discuss the possible metabolic engineering using those genes
involved in dammarene-type ginsenoside saponin biosynthesis in Panax ginseng.

Wed 24, OCT

2Fa08

Advances in PhytoGenomics

Production of plant triterpenoids in engineered yeast○Toshiya MURANAKA, Hikaru SEKI

Abstracts

Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1
Yamadaoka, Suita, Osaka 565-0871, Japan

Plant triterpenoids represent a large and structurally diverse class of natural
products. Among these chemicals, glycyrrhizin, a triterpenoid saponin derived
from underground parts of Glycyrrhiza plants (G. uralensis and G. glabra,
Fabaceae; licorice), is one the most important crude drugs in the world.
Glycyrrhizin and its aglycon glycyrrhetinic acid exhibit various pharmacologicalactivities, including anti-inflammatory and hepatoprotective activities. Glycyrrhizin
is also known to be 150 times sweeter than sucrose. Therefore, a large amount oflicorice and its extracts are on the world drug market as medicinal materials andsweetening agents, although their production largely depends on the collection ofwild licorice plants, and this has caused a decrease in licorice reserves and anincrease in desertification where it is harvested. Here, we show the engineeringof Saccharomyces cerevisiae to produce glycyrrhetinic acid. This was achievedby redirecting a portion of the native 2,3-oxidosqualene pool to heterologous
pathway leading to glycyrrhetinic acid, by expressing three pathway enzymes,
β-amyrin synthase (bAS), which constructs the triterpene skeleton β-amyrin, andtwo cytochrome P450 monooxygenases (CYP88D6 and CYP72A154) these
perform subsequent oxidations of β-amyrin at positions C-11 and C-30,
respectively. Furthermore we could isolate another type of P450 categorized into
CYP716A subfamily, which functions as multifunctional oxidase in triterpenod
2biosynthesis. CYP716A12 was able to generate oleanolic acid, ursolic acid andbetulinic acid coexpressed with bAS, α-amyrin synthase and lupeol synthase inyeast, respectively. Our results provide proof-of-concept for the engineering of the
production of high-value triterpenoid products in yeasts.
[Keywords] P450, Metabolic engineering, Synthetic biology, Triterpenoid36

Bio-Based Production

Living microbes as selective and efficient redox biocatalysts○Andreas SCHMID, Katja BÜHLER, Mattijs JULSING, Bruno BÜHLER

Wed 24, OCT

2Ap01

Laboratory of Chemical Biotechnology, TU Dortmund University, Germany[Keywords] Whole cell biocatalysis, Multistep reactions, Asymmetric epoxidation,
Microreactors, Productivity

37

Abstracts

Redox biocatalysis enables a large and steadily growing number of industriallyinteresting highly specific reactions. Technically demanding reactions, e.g.
oxyfunctionalizations of hydrocarbons, make use of complex enzyme systems likeoxygenases. Here, the application of living microbial cells is most attractive, as
they allow efficient cofactor regeneration, self-renewal (enzyme synthesis andgrowth), and handling of reactive oxygen species. Furthermore, multiple reactions
can be coupled inside the cell functioning as microreactor. Such whole-cellsystems pose challenges with respect to catalyst optimization and processcontrol. Beside technical aspects such as mass transfer and reaction kinetics, alarge number of concurrent and highly cross-linked biological processes such asrecombinant gene expression, energy metabolism, and toxification have to beconsidered.
With the aim to construct the ideal microbial cell for recombinant oxygenasecatalysis, we follow a systems biotechnology approach also considering technicalimplications. The coupling of specific hydrocarbon oxyfunctionalization andtransaminase catalysis will be presented as an example for orthologous reactionengineering. Finally, three approaches for overcoming the inherent problem ofsubstrate and O2 mass transfer limitations will be introduced: substrate uptake
systems, catalytic biofilms in microreactors and regiospecific, O2-independent,
aromatic hydroxylation.

Wed 24, OCT

2Ap02

Bio-Based Production

Toward biological replacement of petroleum○James C. LIAO

Abstracts

Department of Chemical and Biomolecular Engineering, University of California, Los
AngelesPetroleum is the major source that provides energy and chemicals used today.
Unfortunately, because of its unreliable supply, fluctuation in price, and negativeenvironmental impacts, the need to replace petroleum with alternative sources
has become increasingly important. Current biorefining schemes use plantbiomass or algae to produce ethanol, biodiesel, and chemicals. While these are
currently the leading processes, next-generation biorefinery should consider otheroptions as the starting materials and manufacturing technology. For example,
direct photosynthetic production of fuel or chemicals from CO2 is attractive, as
this scheme bypasses the need for lignocellulose deconstruction and algal lipid
processing. Non-photosynthetic, electricity-powered CO2 reduction to fuel orchemicals further by-pass the need for the need for light exposure surface areasin photo-bioreactor, and represents a feasible option for biochemical and biofuel
production as well as electricity storage. Finally, decomposing waste proteins toammonia and fuels or chemicals provides the possibility of closing the nitrogen
cycle and increase the yield and productivity of biorefining.
[Keywords] Metabolic engineering, Synthetic biology, Biorefining, Photosynthesis

38

Bio-Based Production

Bacillus subtilis cell factory for production of scyllo-inositol
promising for Alzheimer’s disease

Wed 24, OCT

2Ap03○Ken-ichi YOSHIDA1, Chumsakul ONUMA 2, Shu ISHIKAWA 2,
Naotake OGASAWARA 2
1

Department of Agrobioscience, Graduate School of Agricultural Science, Kobe
University
2
Graduate School of Biological Sciences, Nara Institute of Science and Technology(NAIST)[Keywords] Bacillus subtilis, Inositol, Bioconversion, Alzheimer’s disease39

Abstracts

Inositol stands for a class of compounds forming nine stereoisomers throughepimerization of the six hydroxyl-groups. myo-Inositol (MI) is one of the isomers,
most abundant in nature, and supplied cheap from phytin in rice-bran. Some ofthe other isomers are rare and thus very expensive but reported to possessinteresting biological functions for the treatment of diseases difficult to treat. For
instance, another isomer scyllo-inositol (SI) was shown to disperse the highmolecular-weight oligomeric aggregates of beta-amyloid peptide in brain, andthus appears promising for Alzheimer’s disease.
We demonstrated that manipulating the inositol metabolism in Bacillus subtilis
enabled the artificial interconversion among inositol stereoisomers secreted into
the culture medium, materializing a B. subtilis cell factory for simple andinexpensive bioconversion of cheap MI to valuable SI. We investigated the effectof medium components on the SI bioconversion efficiency to find that increased
concentrations of Bacto soytone, one of the major nutrients of the bioconversion
medium, improved the efficiency without significant effect on the cell growth. We
thus successfully defined an optimized medium composition for SI bioconversion,by which almost 50% of MI initially contained in the medium was converted to
SI after 48-h culture of bioconversion. Effects of medium optimization ontranscriptome were examined to understand the reasons for the efficientbioconversion, implying that a global change in metabolic pathways might occur
to fulfill the demand of required coenzyme regeneration.

Wed 24, OCT

2Ap04

Bio-Based Production

Engineering of signal transduction for enhanced production ofbiopharmaceuticals and bioenergy○Jian-Jiang ZHONG1, Yang-Chun YONG2
1

Abstracts

State Key Laboratory of Microbial Metabolism, and Lab. of Molecular Biochemical
Engineering, School of Life Sciences and Biotechnology, Shanghai Jiao Tong
University, 800 Dong-Chuan Road, Shanghai 200240, China
2
Biofuels Institute, School of the Environment, Jiangsu University, 301 Xuefu Road,
Zhenjiang 212013, Jiangsu Province, China

Signal transduction engineering has been successfully applied to industrial andenvironmental biotechnology as illustrated in our previous work. Here we report
new progresses in production of biopharmaceuticals and bioenergy by using thisinteresting approach. In liquid fermentation of Ganoderma lucidum, a famous
traditional medicinal mushroom, which produces interesting antitumor ganoderic
acids, calcium signal transduction was found to be significant to the ganodericacid biosynthesis. A possible model on the effect of external calcium ion on theganoderic acid biosynthesis via calcineurin signal transduction pathway was
proposed. Similarly, in bioactive ginsenoside biosynthesis, signal transduction
under heavy metal elicitation led to higher accumulation of ginsenosides in Panax
ginseng cell cultures. In bacterial world, quorum sensing is a wide-conserved
cell-cell communication system and plays important roles in various biologicalprocesses. But it is yet poorly understood what its role is in a bioelectricity
generation process. In a microbial fuel cell system, which is promising for
simultaneous clean energy generation and pollutant treatment, by manipulation of
rhl quorum sensing expression, we found that Pseudomonas aeruginosa could
use different electron shuttles. Interestingly, the electron shuttle substitution (byphenazines) increased the maximum current output of the rhl overexpressedstrain over the wild-type one. Further investigation is under way.
[Keywords] Bioprocess engineering, Signal transduction engineering, Quorum
sensing, Biomedicine, Bioelectricity40

Bio-Based Production

Synthetic biology for the comprehension of biomolecular networks○Masahiro OKAMOTO1,2

Wed 24, OCT

2Ap05

1

Department of Bioinformatics, Graduate School of Systems Life Sciences, Kyushu
University
2
Synthetic Systems Biology Research Center (SSBRC), Kyushu University[Keywords] Synthetic metabolic engineering, Micro fluid engineering, Synthetic
genetic circuit, Evolutional molecular engineering, Embryological engineering

41

Abstracts

In order to make the paradigm shift from the concept of “watched and analyzed
biology” to that of “synthetic and analyzed or utilized biology”, the innovative
research named Synthetic Biology was started from 2000 in US, such as
designing synthetic genetic circuit by combining known interrelated biomaterials,
realizing a certain bio-functional behaviors such as switch, oscillation in vivo,
designing synthetic metabolic pathways by incorporating enzyme coded genes
from other origins into the cells. However, these attempts have been done on asmall scale and with a trial-and-error method. The objectives of this research
project is to establish the coordination between the fundamental technologies forsynthetic biology in order to comprehend biomolecular networks by integrating thefollowing three missions: 1) design synthetic genetic circuit or metabolic pathway
with using the methods of computational science, 2) construct the circuit in vitro
with using the method of engineering, 3) construct the circuit in vivo or in the cellwith using the methods of molecular biology. In order to construct and control alarge scale of dynamic and complex synthetic genetic circuit or metabolic
pathways, the following fundamental technologies for synthetic biology are
essential: Biochemical Engineering, Embryological Engineering, Molecular
Biology, Evolutional Molecular Engineering, Micro Fluid Engineering, Biomolecular
Chemistry, Simulation Engineering, and Knowledge-based Engineering. In thefirst stage (2–3 years), our mission is to construct dynamic and multi-elements
synthetic genetic circuit, followed by the construction of differentiation-induced
system against stem cell and by the realization of cell factory, in which cells can
produce the target metabolites by themselves according to the cell environment in
the last 2-years. The research project is composed of the following four sub
sections: (A01) fundamental technologies of molecular biology, (B01) fundamental
technologies of engineering, (C01) fundamental technologies of computational
science, and (X01) integrated section of A01, B01 and C01.

Wed 24, OCT

2Ap06

Bio-Based Production

Strategies and technologies towards tailor made fuels from biomass○Jochen BUECHS1, Tobias KLEMENT1, Martin KUNZE1, Gernot JAEGER1,
Sandra WEWETZER1, Frederike CARSTENSEN2, Matthias WESSLING2,
Philipp GRANDE3, Pablo Domínguez DE MARIA3, Antje SPIESS4
AVT - Biochemical Engineering, 2 AVT - Chemical Process Engineering, 3Institute ofTechnical and Macromolecular Chemistry, 4AVT - Enzyme Process Technology,
RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany

Abstracts

1

Fuel and chemical industry is facing a large challenge, as its raw material has tobe shifted from fossil to renewable resources in the coming years. Completely
new logistic concepts and technologies to convert biomass to intermediates andfinally to fuels and chemicals have to be developed. In the centre of excellence“Tailor Made Fuels from Biomass (TMFB)”, funded by the German Research
Foundation, the conversion of different kinds of biomass to intermediates for
subsequent fuels production is investigated. Solid biomass, pre-treated bymechanical means, is directly chemically fractionated or dissolved in ionic liquids
and partially chemically hydrolysed to oligomers. Subsequently, the oligomers canenzymatically be converted to glucose. Enzymes are studied which retain theiractivity even in the presence of high concentrations of ionic liquids. This
combined chemo/enzymatic route is completed in much shorter time thanconventional methods. Another possibility to utilize the chemically fractionatedmaterial is a combined enzymatic hydrolysis coupled with a fermentation process.
The robustness of the fermentation in the presence of the residual chemicals
from the preceding pre-treatment step is crucial for an over-all economic process.
In TMFB the itaconic acid production with the smut fungus Ustilago mayadis isinvestigated. This microorganism is relatively insensitive against different stressfactors. With a wild type strain itaconic acid concentrations of 35 g/l wereobtained. Currently, newly developed recombinant strains are investigated. This
fermentation process is designed to be operated in continuous mode with cellretention, using microfiltration membranes. A new pulsed diafiltration technique isused to prevent membrane fouling.
[Keywords] Lignocellulose, Integrated process, Continuous culture, Cell retention,
Membranes

42

Bio-Based Production

Actinobacteria tyrosinase and its applications to biomaterials○Byung-Gee KIM1,2,3, Nahum LEE1,3, SangHyuk LEE2,3, ChangHyun SONG1,3

Wed 24, OCT

2Ap07

1

School of Chemical and Biological Engineering, Seoul National University, Seoul, Korea
Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea
3
Institute of Bioengineering, Seoul National University, Seoul, Korea
2[Keywords] Tyrosinase, Cross-linking of biomaterials, ortho-Dihydroxylation,
Streptomyces

43

Abstracts

Most Streptomyces species produce and secrete melanin, a dark pigment
involved in the protection against UV rays and reactive oxygen species. The key
enzyme to synthesize melanin is tyrosinase, which uses phenolic compounds andcatechol derivatives. Here, we demonstrate the secreted tyrosinase from
Strepomyces avermitilis involved in not only hydroxylation of catechol derivatives
but also degradation of catechol derivatives. Generally, ortho-hydroxylated catechol
derivatives such as 3’ODI and piceatannol (a kind of stilbene compounds) show
higher antioxidant activity than their corresponding monophenolic compounds. To
produce such ortho-hydroxylated catechol derivatives, their hydroxylation has
been done by whole cell reaction of Streptomyce avermitilis by inhibiting
extracellular tyrosinase activity using catechol as an inhibitior as well as a
reducing reagent. Since the tyrosinase secreted by S. avermitilis has both
functions of monooxygenase and dioxygenase, the kinetics of such hydroxylation
reaction of tyrosinase were elucidated more in detail. We have expressed thetyrosinase (MelC2) from S. avermitilis in E. coli with the help of chaperon GroEL/
GroES and its tyrosinase activity to produce ortho-hydroxylated catechol
derivatives was confirmed. In addition, to enhance the ratio of monooxygenase
function to dioxygenase function of tyrosinase, site directed mutagenesis of thehelper protein melC1 and melC2 were attempted. We have compared theactivities of monooxygenase and dioxygenase of the tyrosinase (MelC2) from S.
avermitilis with those of a commercial tyrosinase from Agaricus bisporus(mushroom) using several substrates, and explained their structures based upon
their substrate specificities. We will discuss on how the structures of tyrosinase
affect their applications to biomaterials.

Wed 24, OCT

2Ap08

Bio-Based Production

Cyberinfrastructure for metabolomics and synthetic biology○Masanori ARITA1,2
1

Abstracts

Department of Biophysics and Biochemistry, Graduate School of Science, The
University of Tokyo
2
RIKEN Plant Science Center

A cyberinfrastructure, i.e., web-based research environment, for metabolomics
and related areas is introduced. When a group of people collaboratively work on
the same scope, web-based archives such as Wikipedia or the Registry ofStandard Biological Parts (http://partsregistry.org/) are ideal information
resources. The advantages of such wiki-based sites are the flexibility in its
management and the less psychological barriers to join the community. The
often-said drawbacks of such archives are the lack of security and of data-miningor visualization services due to unorganized formats or missing description. Since
it is difficult to find a full-time administrator or curator for such collaborative
projects, the latter drawback may be fatal for scientific data. To overcome thisdrawback, a MediaWiki-based data repository is designed and multiple extensions
are prepared in programming languages such as Lua and JavaScript (the original
language of MediaWiki is PHP). We standardize data inputs by using templates,and prepare functional pages so that compiled data are automatically subject tostatistical or mining processes. Although our main focus is mass spectra fromdifferent biological sources, these data can be integrated with variety of otherresources such as compounds, plant species, and gene structures. In this talk,
we introduce several examples of automated data processing and explain benefitsof such open data archives. The website is accessible at http://metabolomics.jp/.
[Keywords] Metabolomics, Database, Mass spectra44Bio-Based Production

Bioprocess engineering on biofuels and bio-based chemicals
production from microalgae

Wed 24, OCT

2Ap09○Jo-Shu CHANG1,2,3, Chun-Yen CHEN2
1

Department of Chemical Engineering, National Cheng Kung University, Taiwan
Center for Bioscience and Biotechnology, National Cheng Kung University, Taiwan
3
Research Center for Energy Technology and Strategy, National Cheng Kung
University, Taiwan
2[Keywords] Microalgae, Mitigation of CO2 emissions, Lipid, Carbohydrate, Protein,
Biofuels, Flue gas, Biorefinery

45

Abstracts

The future resources for food, fuels, and chemicals will potentially come fromocean. One of the major marine resources is microalgae, which have the ability to
fix carbon dioxide at a much faster rate than that of terrestrial plants. The fixed
carbon dioxide is converted to microalgal biomass, which has potential
applications in producing biofuels, animal feed, health food, pharmaceuticals, andother high-value products. Therefore, using microalgae to mitigate CO2 emissions
is a promising strategy for CO2 storage and re-utilization. We have identified
various indigenous microalgae strains that utilize flue gas of a steel-making
factory for growth with excellent CO2 biofixation ability. We are able to adjust thecomposition of resulting microalgal biomass (lipid, carbohydrates, proteins,pigments, etc.) by using different cultivation strategies to meet the needs ofdownstream applications. To make the concept of microalgae industry a reality,
new technologies and engineering approaches are developed; for instance,
outdoor large-scale cultivation, biomass harvesting, product conversion technology,
and so on. Some key technologies required for realizing commercialization of
microalgae-based CO2 emission mitigation and biofuels/chemicals production will
be introduced.

Wed 24, OCT

2Ap10

Bio-Based Production

Systems metabolic engineering –Rational design of microbial cell
factories○Hiroshi SHIMIZU1, Chikara FURUSAWA1,2, Takashi HIRASAWA1, Naoaki ONO3,
Katsunori YOSHIKAWA1, Yoshihiro TOYA1
1

Department of Bioinformatic Engineering, Osaka University
Quantitative Biology Center (QBiC), RIKEN
3
Graduate School of Information Science, Nara Institute of Science and Technology(NAIST)

Abstracts

2

In order to establish sustainable societies, production processes should shift frompetrochemical-based processes to bioprocesses. Recently integration of in silico
and experimental approaches for creation of microbial cell factrories is highly
desired. Recently, on the basis of whole-genome information, the genome-scale
metabolic reaction models (GSMs) of cells have been reconstructed for manyorganisms. Using the GSMs, a reliable prediction of metabolic fluxes is possible
by using Flux Balance Analysis (FBA). The constructed GSMs are applied to
prediction of metabolic fluxes under several environmental conditions and todesign of genetic modification for valuable compounds production. Metabolic flux
analysis (MFA) based on quantification of 13C-labeling patterns of metabolites bymass spectroscopy (MS) is also a powerful tool to quantify fluxes experimentallyin a metabolic network of microorganisms. In order to obtain stress tolerant strain,
experimental evolution and multiomics analysis of evolved strains are alsopresented. These analyses provide deep understanding of cellular processes andupgrade designability of cell factories. Systems metabolic engineering gives riseto new knowledge about metabolic systems that can facilitate engineering ofstrains suitable for industrial applications.
[Keywords] Genome scale model, Metabolic flux analysis, Evolution engineering,Multiomics analysis, Metabolic engineering46

Biomolecular Engineering ~ Young Asian Biotechnologist Prize~

Reaction evaluation and new process design in composting of
biological wastes

Thu 25, OCT

3Fa01○Jingchun TANG1, Arata KATAYAMA 2
1

College of Environmental Science and Engineering, Nankai University

2

EcoTopia Science Institute, Nagoya University[Keywords] Composting, Quinone profile method, Denaturing gradient gelelectrophoresis, Alginate degradation

47

Abstracts

A combination of quinone profile method and denaturing gradient gelelectrophoresis (DGGE) of 16S ribosomal RNA genes was applied to characterize
the microbial community of composting process. A higher quinone content andmicrobial diversity was found during thermophilic composting process ascompared to mesophilic composting process. The mesophilic microbialcommunity was characterized by the predominance of ubiquinones andmenaquinone (MK)-8, which correspond to Proteobacteria and fungi. The
thermophilic microbial community was characterized by the dominance of MK-7 in
the initial period, and increases in the amounts of menaquinones with long andpartially-saturated side chains in the later period, corresponding to Firmicutes andActinobacteria, respectively. DGGE analysis also confirmed that diversity of
microbial communities increased but differently in the two processes. Compostingprocess was also applied to dispose marine seaweed waste. Alginate-degrading
bacterium A7 was isolated from wakame (Undaria pinnatifida) compost andconfirmed to belong to the genus Gracilibacillus by partial 16S rDNA analysis.
Alginase was purified from Gracilibacillus A7 and evaluated for its ability to
produce elicitor-active oligosaccharides. Composting of the Undaria pinnatifida(wakame) seaweed was conducted after inoculation with Gracilibacillus sp. A7
and halotolerant bacterium Halomonas sp. AW4. Inoculation with strains A7 andAW4 resulted in 27.8 and 24.7% degradation of wakame dry mass after 168 h ofcomposting, whereas only 17.5% degradation occurred in the uninoculated
control. Inoculation with A7 resulted in 2.8 times faster degradation of alginate
and 1.2 and 1.6 times higher levels of reducing sugars and unsaturated sugars
than inoculation with AW4. The compost produced from the inoculation with A7
had low plant toxicity as measured by germination experiment. Inoculation of
wakame with alginate-degrading bacteria not only shortened the length ofcomposting but also created seaweed compost with good fertilizer qualities.

Thu 25, OCT

3Fa02

Biomolecular Engineering

Organic solvent stable enzymes○Hiroyasu OGINO

Abstracts

Department of Chemical Engineering, Osaka Prefecture University

Enzymes catalyze specific reactions without producing by-product under mild
conditions. It is possible to construct environmentally-friendly sustainablebioprocesses, when enzymes are used as catalysts for chemical processes.
Especially, chemical processes producing fine chemicals consume much energy
and resource and produce much by-product. Although enzymes are good
catalysts for these processes, enzymes are not stable in the presence of organic
solvents used for production of fine chemicals. By the addition of organic solventsto the enzymatic reaction solution, the solubility of substrates and the reaction
rate can often be markedly increased. In the presence of organic solventshydrolytic enzymes can also perform synthetic reactions, which are the reverse
reactions of hydrolysis. A further advantage of non-aqueous reaction solutions isthat the risk of unwanted microbial contamination is normally very low. Therefore
organic solvent-stable enzymes were developed. First, microorganisms which
produce organic solvent stable enzymes were isolated from natural source. Using
organic solvent stable enzymes, some reasons of organic solvent stability ofenzymes were understood. Furthermore, high stable and high active enzymes
were developed from organic solvent stable enzymes by protein engineering.
[Keywords] Organic solvent, Stability, Tolerance, Lipase, Protease48

Biomolecular Engineering

Engineering of aminoacyl-tRNA synthetases for residue-specificincorporation of amino acid analogues into proteins

Thu 25, OCT

3Fa03○Tae Hyeon YOO
Department of Molecular Science and Technology, Department of Applied Chemistry
and Biological Engineering, Ajou University, Suwon, Korea

In the past several decades, there have been significant advances inunderstanding of protein sequences, structures and functions which in turn haveaided engineering of proteins with novel structures and/or functions. In addition,
recent work from several laboratories has shown that an expanded repertoire ofamino acid analogues can be incorporated into proteins. The recruitment of new
amino acid constituents provides the protein engineering field with new tools andraises the prospects for creating novel proteins. In this talk, I will present a highthroughput screening method for engineering aminoacyl-tRNA synthetases toactivate noncanonical amino acids and show examples how the engineered
enzymes are utilized to study biology, especially to interrogate cellular protein
synthesis.

49

Abstracts[Keywords] Amino acid analogues, Protein engineering, Aminoacyl-tRNA
synthetase

Thu 25, OCT

3Fa04

Biomolecular Engineering

Production of the bio-butanol based on the continuous in situ
fermentation and adsorption (ISFA) technology○Hanjie YING1,2, Yong CHEN2,3, Jinglan WU2,3
1

State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing, China
College of Biotechnology and Pharmaceutical Engineering, Nanjing University ofTechnology, Nanjing, China
3
National Engineering Technique Research Center for Biotechnology, Nanjing, China

Abstracts

2

Since butanol is an important chemical and a potential energy-substitute,
production of butanol based on the biological methodology has being obtained
comprehensive attention from academic and industrial institutions all of the world.
However, similar to the other bulk chemicals manufactured by biotechnology, thepreparation of bio-butanol nowadays exists several problems, i.e., use of thefoodstuffs as substrate, low economic efficiency, high energy-consumption as
well as high environmental pollution.
This report focuses on the following two questions. The corresponding solutions
were proposed and the main results were presented subsequently. Firstly,
production of bio-butanol commonly suffered from poor strain tolerance, low
productivity and butanol yield, which cause low butanol titers, large investment inequipment and consumption of raw materials. A novel method for cell
immobilization by surface adsorption and strategies of cofactor manipulation weredeveloped to address these problems. The fermentation time was significantly
shortened from 40–50 h to 10–14 h while the butanol titer was increased from
12–13 g/L to 15–16 g/L, which resulted in a three-fold increase in productivity.
Secondly, due to the high energy-consumption and large amount waste-waterdisposal in the traditional distillation process, an innovative in situ productrecovery (ISPR) technology was developed incorporation with the processsimulation to recovery bio-butanol from the fermentation broth. As a result, thebio-butanol concentration was improved from 1.2% to 15–20%, the energyconsumption was reduced 25–30%, and the waste-water disposal was decreasedaround 50%, which provided the fundamental technology to realize the highefficiency production of bio-butanol in an industrial scale.
[Keywords] Bio-butanol, Cofactor manipulation, Adsorption immobilization, ISPR

50

Biomolecular Engineering

Functional properties of diet ginger (Zingiber officinale Roscoe,
Zingiberaceae) for antioxidant, anti-pigmentation, anti-cancer,
anti-bacterial and wound repair

Thu 25, OCT

3Fa05

Pei-Fang WU, Yi-Ting CHOU, ○Hui-Min WANG
Department of Fragrance and Cosmetic Science, Kaohsiung Medical University,
Kaohsiung, Taiwan, ROC

Ginger, the rhizomes of the herb Zingiber officinale Roscoe (Zingiberaceae), is aperennial herb and is used widely as a spice throughout the world. There aremany species of ginger, mainly cultivated in tropical countries of Asia. We
published that biologically active compounds found in diet ginger has been shownto have a number of pharmacological activities including anti-oxidant, antipigmentation, anti-cancer, anti-bacterial and to reduce the metabolic syndrome. It
has important economic value and a wide range of pharmacological effects.

[Keywords] Ginger, Anti-oxidant, Anti-pigmentation, Anti-cancer, Anti-bacterial

51

Abstracts• Chung-Yi Chen, Chien-Chih Chiu, Chi-Peng Wu, Yi-Ting Chou, Hui-Min Wang*
Enhancements on skin cell proliferations and migrations via 6-dehydrogingerdione
2012, submitted to PLoS ONE• Chung-Yi Chen, Kuo-Chen Cheng, Andy Y Chang, Ying-Ting Lin, You-Cheng
Hseu, Hui-Min Wang*
10-Shogaol, an Antioxidant from Zingiber officinale for Skin Cell Proliferationand Migration Enhancer
International Journal of Molecular Sciences. 13, 1762-1777, 2012
• Hui-Min Wang, Chung-Yi Chen, Hsi-An Chen, Wan-Chun Huang, Wei-Ru Lin,
Tun-Chieh Chen, Chun-Yu Lin, Hsin-Ju Chien, Po-Liang Lu, Chiu-Mei Lin, YenHsu Chen*
Zingiber officinale (ginger) Compounds with Tetracycline Have Synergistic
Effects against Clinical Extensively-Drug Resistant Acinetobacter baumannii
Phytotherapy Research. 24, 1825-1830, 2010
• Bing-Hung Chen, Pei-Yu Wu, Kuan-Ming Chen, Tzu-Fun Fu, Hui-Min Wang*,
Chung-Yi Chen*
Antiallergic Potential on RBL-2H3 Cells of Some Phenolic Constituents of Zingiber
officinale (Ginger)
Journal of Natural Products. 72, 950–953, 2009

Thu 25, OCT

3Fa06

Biomolecular Engineering

Enzyme engineering with the aid of ionic liquids○Noriho KAMIYA1,2
1

Center for Future Chemistry, Kyushu Univeristy
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University

2

Abstracts

For more than a decade, ionic liquids (ILs) have been extensively exploited as
new reaction media for biocatalysts. The great potential of ILs is well recognizedbecause the physicochemical characteristics can be tuned by altering the pair ofcation/anions or by introducing functional substituents into the IL components.
Indeed, a range of enzymes has proven to exhibit catalytic activity in ILs with little
or only a few percent water content. Recently, the potential of ILs as uniquesolvents for the pretreatment of lignocellulosic biomass has been focused, which
opens a new avenue to the utilization of renewable bioresources for enzyme
engineers. Previous reports including ours clearly indicate that ILs can be
employed not only as simple reaction media but also as potent chemicals foraltering substrate characteristics in enzymatic catalysis. Here we report ourongoing efforts on how to utilize ILs in enzymatic saccharification of cellulosic
biomass. The latest trial for the construction of new IL-based reaction media forbiotransformation will be presented as well.
[Keywords] Ionic liquids, Lignocellulosic biomass, Solvent engineering

52

SBJ/JST Joint Symposium I -General Session-

Introduction of the JST CREST/PRESTO program research area
“Creation of basic technology for improved bioenergy production
through functional analysis and regulation of algae and otheraquatic microorganisms”

Thu 25, OCT

3Ap01○Tadashi MATSUNAGATokyo University of Agriculture and Technology[Keywords] Biofuel, Algae, Aquatic microorganisms, Omics, Metabolic pathways

53

Abstracts

Biological energy production from solar energy has been a human dream for along time. Ethanol produced from corn and sugar cane by yeast has already beencommercialized as a biofuel. However, the use of these sources for bioethanol
competes with food production. Therefore, the use of non-edible crop components
and wood waste has become an important research target. Recently, biofuel
production using algae and other aquatic microorganisms has attracted attention,
because these organisms cannot only be used to produce ethanol but also to
produce biodiesel fuel and other hydrocarbons. Use of the marine environment is
an important advantage because the oceans cover 70% of the Earth. This
Research Area aims to create innovative technologies for bioenergy production
through the functional analysis of algae and other aquatic microorganisms basedon recently developed “omics” technologies, using marine and freshwaterorganisms and specifically focuses on the production of biofuels, including
biodiesel fuel, light oil (alkane and alkene), ethanol, methane, hydrogen, etc., aswell as lipids and sugars that can be converted to biofuels. For that purpose,as well as the collaboration among researchers from various fields, including
marine biotechnology, phycology, microbiology, bioinformatics, marine biology,
biochemistry, genetic engineering, plant physiology, chemistry, chemicalengineering, etc., challenging projects with creative ideas from new and active
researchers are necessary. This Research Area is jointly operated with multiple
CREST and PRESTO researchers. This Research Area is expected to generatebasic technology for the low cost production of biofuels. The use of fossil fuels
including crude oil will be decreased with the use of new biofuel production
technologies. Furthermore, the establishment of new metabolic pathways in algae
and other aquatic microorganisms that lead to the production of chemicals,
including precursors of plastics, may change the current dependency on oil
among chemical industries. Active mutual information exchange of each research
progress enhances synergy between these projects and promotes further
achievements, like new technologies within five to ten years through large-scale
pilot plant experiments upon completion of this Research Area.

Thu 25, OCT

3Ap02

SBJ/JST Joint Symposium I -General Session-

Can biotechnology address the global energy crisis?
○Jonathan D. TRENT 1,2
1

Bioengieering Branch, NASA Ames Research Center
Biomolecular Engineering, University of California Santa Cruz

Abstracts

2

There are claims that biofuels produced by microalgae can provide a sustainable
substitute for fossil fuels. Indeed, some known species of microalgae are fast
growing and provide the highest yields of biodiesel. There are however,
formidable challenges to meet the demands of large-scale production within theconstraints of economics and without competing with agriculture for freshwater,
fertilizer, or land. To meet these challenges, we have proposed a system called
OMEGA (Offshore Membrane Enclosures for Growing Algae). In the OMEGA
system, oil-producing freshwater algae are grown in flexible, clear plastic
photobioreactors (PBRs) attached to a floating infrastructure anchored offshore ina protected bay. Domestic wastewater (sewage) and CO2 from coastal facilities
provide water and nutrients. The surrounding seawater controls the temperatureinside the PBRs. The salt gradient between seawater and wastewater kills algaethat escape from the system and drives forward osmosis to concentrate nutrientsand facilitate algae harvesting. The OMEGA infrastructure supports traditional
aquaculture and alternative energy (solar, wave and wind). Integrating algae
cultivation, wastewater treatment, CO2 sequestration, aquaculture, and alternative
energy supports the economics of the system as a whole. By using domestic
wastewater for water and fertilizer and by operating offshore, OMEGA does notcompete with agriculture for water, fertilizer, or land. By treating wastewater,
sequestering CO2, and creating a “floating reef,” OMEGA has a positive impact on
the local environment. Progress into the feasibility of OMEGA will be discussed.
[Keywords] Biofuels, Algae, Photobioreactor, Wastewater treatment, Aquaculture54SBJ/JST Joint Symposium I -General Session-

Food and fuel from microalgae: Integration of photosynthesis andphotovoltaics allows to achieve a positive energy balance of algae
biomass production

Thu 25, OCT

3Ap03○Mario R TREDICI
Dipartimento di Biotecnologie Agrarie, Università degli Studi di Firenze[Keywords] Microalgae, Energy return on investment, Green wall panel (GWP),
Photobioreactor, Photosynthesis and photovoltaics

55

Abstracts

In spite of the potential of microalgae, no company seems to possess a maturetechnology able to produce algae biomass at low cost and compete withtraditional sources of food, feed and fuel. The high capital and operating costs of
microalgae farming, the non-sufficiently positive energy balance and the not yet
established sustainability still prevent the development of this technology tocommercial scale. But new designs are emerging, which may contribute to closing
the gap between algae feedstock and current alternatives.
Among the barriers that need to be cleared before fuel, feed and food from algae
may become a reality, there is the sustainable cultivation of selected strains atlarge scale and the low EROI (Energy Return on Investment) of the process. The
main reasons for the preference of photobioreactors (PBR) over ponds for algae
cultivation is that they provide a close and more controllable environment, thus
limiting the risk of contamination and ensuring a higher and more sustainableproduction. However, no PBR design has been developed and tested at the (large)
scale necessary for a complete economic and energetic evaluation of theprocess, and at pilot scale PBR show a lower EROI compared to ponds.
The Green Wall Panel (GWP), a low-cost, flat PBR developed at the University of
Florence and commercialized by Fotosintetica & Microbiologia S.r.l. (Italy), needs
about 350 KJ m-2 d-1 to operate (mostly for mixing and cooling). In the summer,with an average productivity of 20–25 g m -2 d-1 (equivalent to an energy output of
about 400 KJ m-2 d-1), the biomass production process in this system achieves an
EROI near unity, while an EROI of more than three would be necessary. An
integrated GWP has been recently developed to reduce energy costs andenhance the EROI. Called SOLO, this stand-alone system uses part of theimpinging photons for photovoltaic generation of about 1 MJ m -2 d-1 of electricenergy, which are sufficient to cover all the needs of the process (including
cultivation, harvesting and drying). In a particular application of the SOLO tested
outdoors, it has been shown that a significant fraction (30–50%) of the impinging
photons can be diverted to the photovoltaic elements without reducing algal
productivity.
• Rodolfi L., Chini Zittelli G., Bassi N., Padovani G., Biondi N., Bonini G., Tredici M.R.
(2009) Microalgae for oil: strain selection, induction of lipid synthesis andoutdoor mass cultivation in a low-cost photobioreactor. Biotechnol. Bioeng. 102:
100-112.
• Tredici M. R. (2010) Photobiology of microalgae mass cultures: understanding
the tools for the next green revolution. Biofuels 1(1): 143-162.

Thu 25, OCT

3Ap04

SBJ/JST Joint Symposium I -General Session-

Biorefinery of microalgae○René H. WIJFFELS

Abstracts

Wageningen University, Bioprocess Engineering, P.O. Box 8129, 6700, EV
Wageningen, the Netherlands

Our mission is to develop sustainable biobased production strategies in whichphototrophic microalgae are used for a single step conversion of light energy intofunctional products without depletion of natural resources (1,2).
Microalgae are considered one of the most promising feedstocks for sustainableproduction of commodities such as food, feed, chemicals, materials and biofuels.
The technology for production is still immature, but if developed it is expectedthat biomass can be produced at commercial scale for a cost price less than
0,68 €/kg dry biomass (3). If the different biomass components are collected
the total value for commodities in algal biomass is higher than 1.65 €/kg dry
biomass (4).
Recently, we started the project AlgaePARC (www.AlgaePARC.com). AlgaePARC
is a pilot facility with which we intend to bridge the gap between basic researchand demonstration projects. In AlgaePARC we will compare state of the art
technologies and develop new reactor concepts and process control strategies.
As a follow up projects are started to develop technology for disruption of algal
biomass and fractionation of the biomass into different compounds withmaintenance of their functionality. The technologies we develop should be mild,
effective with respect to extraction efficiency and product purity and require aminimal amount of energy and costs. Algal biomass produced at AlgaePARC will
be used for biorefining.
“Biorefinery” is a facility that integrates cell disruption, extraction, conversion andseparation technologies of biomass. The biorefinery strategy is analogous totoday’s petroleum refinery, in which multiple fuel products and chemicals areproduced from crude petroleum. Biorefinery includes the selective isolation of
products (proteins, carbohydrates, lipids) from crude biomass.
The co-production of multiple products from microalgae is a new challenge.
Research and development needs to be done to explore scalable mild cell
disruption and extraction technologies that keep the functionality of the different
cell components.
1. Wijffels R.H., Barbosa M.J. (2010) Science, 379: 796-799
2. Wijffels R.H., Janssen M., Barbosa M.J. (2011) Microbial Biotechnol, 4: 132134
3. Norsker N.H., Barbosa M.J., Vermuë M.H., Wijffels R.H. (2010) Biotechnology
Advances, 29: 24-27
4. Wijffels R.H., Barbosa M.J., Eppink M.H. (2010) Biofuels, Bioproducts, &Biorefining, 4: 287-295[Keywords] Microalgae, Biorefinery56

Japan-Korea Biomass Session

Improvement of ethanol yield from xylose by breeding of industrial
yeast

Thu 25, OCT

3Ap05○Haruyo HATANAKA, Tomohide MAEDA, Yuri YASUE, Kazue NISHIYAMA
Suntory Business Expert Limited, Institute for Microbial Science[Keywords] Bioethanol, Xylose, NAD kinase, Glycerol channel

57

Abstracts

Xylose is one of main sugars abundant in biomass like switch grass, corn cobs,
rice straws and wood chips. A xylose fermenting yeast has been constructed byrecombining genes of xylose reductase (XR) and xylitol dehydrogenase (XDH)from Pichia stipitis, and a gene of xylulose kinase (XKS) from Saccharomyces
cerevisiae under the control of strong promoter. When this recombinant yeast
ferments xylose to ethanol, substantial quantities of xylitol and glycerol are also
produced as by-products, resulting in less efficient ethanol production, comparedwith that from glucose. It is considered that cofactor imbalances might be thereason for the inefficient ethanol production, because XR requires NADPH as a
cofactor, while XDH uses NAD as a cofactor. S. cerevisiae uses NAD for theoxidative reactions and NADPH for the reductive reactions in order to synthesis
biomass. S. cerevisiae reoxidizes NADH to NAD by the reaction to make glycerol
under anaerobic condition. S. cerevisiae has three NAD(H) kinases, Utr1p andYef1p in the cytosol, and Pos5p in the mitochondria. These NAD kinases havedifferent preferences for cofactors as substrates. In this study, we tested whether
the deletions or overexpressions of these NAD kinase genes change the cofactor
balance and improve the ethanol yields from xylose. The strain disrupted UTR1
and YEF1 showed the highest ethanol yield and produced the lowest xylitol andglycerol among the bred strains. It indicates that the shortage of NAD is one of
the reasons for a high accumulation of xylitol. Moreover, we found that xylitol was
excreted by glycerol channel, Fps1p, at some extent, and the disruption of Fps1p
decreased xylitol production and resulted in higher ethanol yield. Industrial yeast
often has the ability of high ethanol production, or high stress tolerance. However
most of them are multiploid and have no spolulation ability. We tried to delete
each set of UTR1, YEF1 and FPS1 from one industrial diploid strain by using amarker recycling system, and confirmed these breeding for the fermentation fromxylose were also available for industrial yeast of diploid.

Thu 25, OCT

3Ap06

Japan-Korea Biomass Session

Applications of microbial factory technology to production ofbioethanol from cellulosic biomass
Jung-Hyun JO1, Sang-Min JUNG1, Yong-Cheol PARK 2, Yong-Su JIN3,○Jin-Ho SEO1
1

Abstracts

Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742,
Republic of Korea
2
Department of Advanced Fermentation Fusion Science and Technology, Kookmin
University, Seoul 136-702, Republic of Korea
3
Department of Food Science and Human Nutrition, University of Illinois at
Urbana-Champaign, Urbana, IL 61801, USA

Microbial factory technology (MFT) is composed of engineering of the key
metabolic pathways, enforcement of cofactor biosynthetic capacities and global
optimization from genetic modification at a gene level to fermentative strategies at
a process scale. MFT has been used to produce bioethanol from cellulose
biomass by metabolically engineered Saccharomyces cerevisiae.
As cellulosic biomass hydrolyzates are composed pentoses (C-5 sugar) andhexoses (C-6 sugar), economic production of cellulosic bioethanol requires
utilization of both sugars, primarily glucose and xylose. Wild type Saccharomyces
cerevisiae cannot convert xylose to ethanol because the yeast does not have axylose metabolizing activity. One of the key problems with using the recombinant
S. cerevisiae transformed with the Scheffersomyces stipitis derived genes coding
for NADPH-dependent xylose reductase (XR) and NAD+-dependent xylitol
dehydrogenase (XDH) as a cellulosic bioethanol producer includes the cofactor
mismatch between XR and XDH.
Protein engineering was employed to make NADH-preferring XR and NADP+dependent XDH. A synthetic isozyme system was generated by expressing bothwild and mutant XR or both wild and mutant XDH and characterized for the ability
of xylose metabolism in an effort to construct a metabolic highway for xylose
utilization.
S. cerevisiae has transporters (proteins responsible for sugar transport) with highaffinity for glucose relative to xylose. Xylose transport is strongly inhibited by thepresence of glucose in the fermentation broth. A mixture of cellobiose and xylose
was used instead of a mixture of glucose and xylose to avoid competitivetransport between glucose and xylose. A recombinant S. cerevisiae wasconstructed for simultaneous utilization of both cellobiose and xylose byintroducing the two genes encoding a cellobiose-specific transporter and
β–glucosidase able to hydrolyze cellobiose to glucose. Engineering optimization
was done to determine the optimum profiles of cellobiose and xylose withfermentation time for the maximum productivity of ethanol.
[Keywords] Cellulosic biomass, Bioethanol, Recombinantcerevisiae, Synthetic isozyme system, Sugar transport

58

Saccharomyces

Japan-Korea Biomass Session

Potential in bioethanol production from waste paper sludge inpulp-based biorefineryThu 25, OCT

3Ap07○Enoch Y. PARK1,2, Joni PRASETYO1, Kazuya NARUSE2, Tatsuya KATO2
1

Laboratory of Biotechnology, Graduate School of Science and Technology, Shizuoka
University
2
Laboratory of Biotechnology, Faculty of Agriculture, Shizuoka University[Keywords] Paper sludge, Biorefinery, Bioethanol, Cellulase, Saccharification

59

Abstracts

Ethanol production from waste papers or paper sludge (PS) of pulp industry is
considered to be the most appropriate way to the biorefinery process, because it
does not require any delignification process. In this study, cellulase production
from cellulase producer, saccharification of paper wastes using produced
cellulase and ethanol production from hydrolysate of waste papers were
investigated. Especially, a simultaneous saccharification and fermentation (SSF)
was conducted using a cellulase produced from PS by the hyper cellulase
producer, Acremonium cellulolyticus C-1 for PS saccharification, and athermotolerant ethanol producer Saccharomyces cerevisiae TJ14 for ethanol
production. Using cellulase of PS origin minimizes biofuel production costs,
because the culture broth containing cellulase can be used directly. When 50 g
PS cellulose/l was used in SSF, the ethanol yield was 0.23 g ethanol/g PS
cellulose, and was two times higher than that in separated hydrolysis andfermentation (SHF). Cellulase activity throughout in SSF remained around 50% to
the initial activity. The ethanol yield in the range from 50 to 150 g PS cellulose/l in
SSF was 0.22–0.24 g ethanol/g PS cellulose. Ethanol production was 40 g/l
under the condition of 161 g/l of PS cellulose using 15 (filter paper unit/g PS
cellulose) at 80 h in SSF. This indicates that the utilization of PS as raw material
for bioethanol production offers a good prospect. On the other hand, one-pot
bioethanol production including both productions of cellulase and ethanol was
conducted using the hyper cellulase producer, Acremonium cellulolyticus C-1 andan ethanol producer S. cerevisiae with Solka flock (SF), crystal cellulose as asole carbon source. Cellulase activity remained 8–12 FPU/ml during one-pot
process. Ethanol concentration increased with the SF concentration. When 50–
300 g SF/l was used in 500 ml Erlenmeyer flask scale, the ethanol concentrationand yield based on initial SF were as 8.7–46.3 g/l and 15.4–17.5% (g ethanol/gSF), respectively. This indicates that one-pot ethanol process, producing cellulase
and ethanol with two different microorganisms, is an option for bioethanol
production.

Thu 25, OCT

3Ap08

Japan-Korea Biomass Session

Application of Metabolomics to high resolution phenotype analysis○Eiichiro FUKUSAKI

Abstracts

Department of Biotechnology, Graduate School of Engineering, Osaka University.

Metabolomics is applicable without genome sequence information. Therefore,
Metabolomics would be powerful analytical tool for commercially available plantand microorganisms. According to the above context, Metabolomics means
exhaustive relationship analysis between ‘data matrix of metabolic profile’ and‘data matrix of performance of samples’. Among those operation of Metabolomics,
metabolic fingerprinting, in which metabolic profile is used as fingerprint for
samples’ performance prediction and classification, is one of the most important
application of Metabolomics. On this occasion, practical operation of
Metabolomics for high resolution phenotype analysis would be presented. To
reveal a power of Metabolomics, a possibility of Metabolomics based prediction of“Life span”, which is highly integrated and complicated quantitative phenotype,was studied to find strong relationship between metabolome and lifespan bymeans of PCA. Orthogonal projection on latent structure (OPLS) based model
can predict yeast life span using metabolome as fingerprint. Using information ofprincipal component vector in PCA life span related metabolites were nominatedand life span related genes were speculated. The result implied Metabolomics
would be applicable for high resolution analysis of a complicated quantitative
phenotype, and metabolome information might be useful for strategicenhancement of useful quantitative phenotype for biomass production.
[Keywords] Metabolomics, Metabolic profiling, Metabolome, GC/MS, Multi variate
analysis60

Japan-Korea Biomass Session

Biological production of 3-hydroxypropionic acid from glycerol○Sunghoon PARK

Thu 25, OCT

3Ap09

School of Chemical and Biomolecular Engineering, Pusan National University[Keywords] 3-Hydroxypropionicengineeringacid,

1,3-Propanediol,

Glycerol,Metabolic61

Abstracts

For every 9 kg of biodiesel produced, about 1 kg of crude glycerol is left behind
which poses a challenge and opportunity to find the best use of it. Here we
present the production of a valuable platform chemical, 3-hydroxypropionic acid
(3-HP), from glycerol using genetically engineered Escherichia coli and Klebsiellapneumoniae strains. In one approach, we screened and expressed various
aldehyde dehydrogenases (ALDHs) along with glycerol dehydratase (GDHt) in E.
coli and demonstrated that a recombinant bearing GDHt and an ALDH produced
3-HP above 50 g/L in 72 h with an average yield of 33% on glycerol. To improve
the yield, the recombinant E. coli strain was further developed by disrupting thegene glpK (glycerol kinase), ldhA (lactate dehydrogenase), frd (fumarate
reductase AB). The resting cells of the recombinant mutant produced 3-HP withan average yield of 80.5% on glycerol. In the other approach, NAD+-dependent
gamma-glutamyl-gamma-aminobutyraldehyde dehydrogenase (PuuC) of K.
pneumoniae DSM 2026 which oxidizes 3-hydroxypropionaldehyde to 3-HP washomologously overexpressed in various K. pneumoniae DSM 2026 and a newly
isolated K. pneumoniae J2B. The K. pneumoniae strains have been engineered
to different genetic background such as ∆dhaT, ∆yqhD, ∆ldhA ∆adhE, ∆glpK. etc
individually and in combination. Recombinant K. pneumoniae could successfully
produce 3-HP and 1,3-propanediol (1,3-PDO) separately or simultaneously. This
talk will summarize our recent work on the production of 3-HP and 1,3-PDO fromglycerol.

Thu 25, OCT

3Ap10Japan-Korea Biomass Session

Biodiesel production by enzymatic process and the utilization of
crude glycerol
Hah Young YOO, Laxmi Prasad THAPA, Ja Hyun LEE, Sang Jun LEE,
○Seung Wook KIM

Abstracts

Department of Chemical and Biological Engineering, Korea University, Seoul, Korea

Currently environmental, economical and security issues demanded alternative
energy sources of fossil fuel and petroleum products. Therefore, nowadays
biofuels like biodiesel and bioethanol became more attractive sources of energy
due to its renewable and environmental benefits. In this work, advanced
enzymatic immobilization method and optimal reaction conditions for biodiesel
production were investigated. Non-edible oil sources, including waste oil were
utilized as biodiesel feedstock for environmental friendly production process.
Circulation and continuous processes by using co-immobilized lipase were
investigated for the improvement of biodiesel production in a packed-bed reactor.
Secondly, crude glycerol which is by-product of biodiesel production was used forcephalosporin C (CPC) and bioethanol production. Acremonium chrysogenum
M35 showed high levels of differentiation of hyphal fragments into arthrospores
when cultured in the presence of glycerol. The glycerol utilization led tosignificantly higher CPC production as compared to rice oil.
Similarly, bioethanol was produced continuously from Enterobacter aerogenes
using immobilized cells in bioreactor. To enhance the production of bioethanolfrom crude glycerol we optimized various process parameters by using statisticalmethod. We also developed a glycerol utilizing Escherichia coli BL21 (DE3) strain
for the production of bioethanol in the presence of ethanol producing gene set ofEnterobacter aerogenes.
The results presented here suggest that low-priced glycerol takes the place of
natural oil and methionine for CPC and bioethanol production in the industrial
scale.
[Keywords] Biodiesel, Bioethanol, Cephalosporin C, Enzyme immobilization

62

SBJ/JST Joint Symposium II -Biorefinery-

Understanding and altering the unique metabolism ofhyperthermophilic archaea

Fri 26, OCT

4Aa01○Haruyuki ATOMI1,2, Kunio MIKI2,3, Tadayuki IMANAKA 2,4
1

Department of Synthetic Chemistry and Biological Chemistry, Graduate School ofEngineering, Kyoto University
2
CREST, Japan Science and Technology Agency (JST)3
Department of Chemistry, Graduate School of Science, Kyoto University
4
Department of Biotechnology, College of Life Sciences, Ritsumeikan University[Keywords] Archaea, Hyperthermophile, Metabolism, Regulation

63

Abstracts

Archaea represent the third domain of life and display unique biological properties
not found in Bacteria and Eucarya. Their membrane lipids consist of isoprene
molecules covalently bound to glycerol molecules through ether bonds. The
methanogens, chemoautotrophs that rely on the conversion of hydrogen andcarbon dioxide to methane as an energy source, are all members of the archaea.
Our studies are mainly focused on the hyperthermophilic archaeon,
Thermococcus kodakarensis. This organism is an obligate anaerobe andheterotroph, and grows at extremely high temperatures with an optimum at 85˚C.
We have determined its genome sequence and have also established a genetic
system through which we can disrupt, insert and modify genes of interest. The
genetic system allows us to directly examine and confirm gene function in vivo.
We are presently interested in the unique metabolism of this archaeon and its
regulation. The organism can degrade a variety of sugar polymers with enzymes
and pathways distinct to eukaryotes/bacteria. T. kodakarensis can utilizeelemental sulfur as a terminal electron acceptor, but can also dispense electrons
via hydrogenases, resulting in the generation of molecular hydrogen. Here we will
present our recent findings on the unique metabolism of this organism, giving
attention to enzymes/pathways related to glycolysis/gluconeogenesis, hydrogenproduction, pentose metabolism and coenzyme A biosynthesis. We will alsopresent some initial attempts to alter the metabolism of T. kodakarensis utilizing
the gene manipulation system developed in this archaeon.

Fri 26, OCT

4Aa02

SBJ/JST Joint Symposium II -Biorefinery-Blue Ocean for Blue Biotechnology – Marine Bioenergy Production○Choul-Gyun LEE1,2
1

Department of Biotechnology, Inha University, Incheon 402-751, Korea
Korea National Consortium on Marine Bioenergy Research

Abstracts

2

Algal biotechnology is drawing increasing interest due to its potential as a sourceof sustainable clean liquid energy. Microalgal biotechnology has also shown
enormous potentials in valuable pharmaceuticals, pigments, carbohydrates, andother fine chemicals. Its application has been extended to the areas of wastewater
treatment and agriculture. Recent development in various algal biotechnology
found microalgal mass culture can be a useful solution in treatment of wastewater,
fixation of carbon dioxide and production of biofuel. However, despite all theseadvantages of microalgal biofuels, there are quite a number of challenges toovercome before economic production of microalgal biofuel can be achieved: (i)finding/constructing algae strain(s) suitable for mass culture and for wide range ofclimate; (ii) maximizing solar conversion efficiency in mass culture; (iii) achieving
both high oil content and high productivity in mass culture; (iv) designing andengineering of cost effective sustainable mass culture systems; (v) harvesting
microalgae and extracting microalgal oils with minimal use of energy; (vi) finding
cheap (and renewable) sources for methanol and nutrients (such as phosphateand nitrate). Most of the culture systems and bioprocesses available today would
be suitable for the products that cost over $20 USD/kg. Bioenergy must be
produced much cheaper than most of the biologically-driven products. One of thepossible solutions for some of these challenges is mass culturing microalgal inlarge ocean area. All the opportunities and challenges of microalgal biofuels will
be discussed. Possible ranges for different biofuels will be estimated using the
maximum energy conversion efficiency from Sun to biomass and/or biofuels.
Then, the feasibility of various bioenergy production technologies will becompared by economic analysis.
[Keywords] Marine microalgae, Ocean culture, Photobioreactor, Microalgal
biodiesel64SBJ/JST Joint Symposium II -Biorefinery-

Establishment of innovative technology to create new microalgal
strains increasing biofuel production by polyploidization andheavy-ion beam irradiation

Fri 26, OCT

4Aa03○Shigeyuki KAWANO1,2, Koichi WATANABE1,2, Shuhei OTA1,2
1

Department of Integrated Biosciences, Graduate School of Frontier Sciences, The
University of Tokyo
2
CREST, Japan Science and Technology Agency (JST)[Keywords] Chlorella,
Astaxanthin

Haematococcus,

3D

reconstruction,

Sulfur-deficient,

65

Abstracts

To make the biofuel production using microalgae practicable, it is necessary to
breed their strains which can be mass-produced just like grains and horticultural
crops. However, there was no conception of breeding in the microalgae so far, or
their genomes have not been decoded in most of species. In the present study,
the microalgae irradiated with the heavy-ion beams, which have the successful
results by the selective breeding of horticultural crops, will be selected and bredbased on the quantitative data concerning their forms. We aim for theestablishment of the breeding scheme, which is innovative, advanced andspecialized in microscopic algae, based on complete genome information.
The green microalgae, Chlorella and Haematococcus, have been focusing on
their own natural height-productivity of starch and oil. Chlorella species undersulfur-deficient conditions promoted transient accumulation of starch followed bya steady increase in lipid storage. Transmission electron microscopy (TEM)
indicates an increase and decrease in starch granules and subsequentenlargement of lipid droplets under sulfur-deficient conditions. They accumulate
1.5–2.7-fold higher amounts of starch and 1.5–2.4-fold higher amounts of lipid
under sulfur-deficient conditions than under sulfur-sufficient conditions.
Haematococcus pluvialisi is known by accumulating “astaxanthin” in the stressfulenvironment like light and oligotrophic conditions, and it is used widely in thebiomass industry in recent years. Ultrastructural 3D reconstruction based on
300–350 serial sections for TEM has applied to analyze the dynamics ofastaxanthin-accumulation and chloroplasts-degradation during encystment under
the continuous light. There are no developing chloroplasts but many large starchgrains surrounded by small fragmented residues of the thylakoid membrane. They
decrease gradually whereas many large oil drops, which seemed to accumulate
astaxanthin, appeared in the cytoplasm outside the chloroplast. They occupied
finally not less volume than 52% of cell. We are now isolating many mutant strainsof Chlorella and Haematococcus by polyploidization and heavy-ion beamirradiation.

Fri 26, OCT

4Aa04

SBJ/JST Joint Symposium II -Biorefinery-

New dimensions in microalgae biomass production for bioenergy○Otto PULZ
IGV, Nuthetal, Germany

Abstracts

Recent developments in mass production of microalgae as phototrophic
microorganisms are demonstrated.
In recent years there arose an urgent need for new solutions for phototrophic
biotechnology out of the development of technologies for carbon capture (CO2)and bioenergy.
The most widespread technology for the mass production of microalgae inraceway ponds is with a productivity of 15–20 g/m2×d not productive enough to fit
in the demands of CO2-emitting companies like power plants.
Two goals are in focus for the emerging phototrophic technologies:- high productivities in mass cultivation (up to 80 g/m2×d)- low cost area demand
The key issue is to find effective ways to introduce light into photobioreactors.
The most important designs of photobioreactors with their R&D demands aredescribed.
[Keywords] Microalgae, Mass cultivation, Photobioreactors

66SBJ/JST Joint Symposium II -Biorefinery-

Development of sustainable biorefinery based on microalgae○Akihiko KONDO1,2

Fri 26, OCT

4Aa05

1

Department of Chemical Science and Engineering, Graduate School of Engineering,
Kobe University
2
Core Research for Evolutional Science and Technology (CREST), Japan Science andTechnology Agency (JST)[Keywords] Bio-refinery, Bio-ethanol, Microalgae

67

Abstracts

Bio-refinery, the utilization of biomass as the starting material for the productionof fuels and chemicals, has received considerable attention in recent years.
Polysaccharides present in lignocellulosic materials are of great interest asalternative feedstocks for second-generation ethanol production, because they
are abundant, inexpensive, and renewable sources of sugars for microbial
fermentation. However, the use of terrestrial plants will soon be limited by theunavailability of vegetated regions on earth and fresh water required for growing
plants. Microalgae have attracted attention as an alternative feedstock because
they convert solar energy to algal biomass more efficiently than higher plants.
Furthermore, microalgae grown in an aquatic environment provide additional
benefits of year-round cultivation without requiring arable land. In particular, theuse of salt-tolerant microalgae grown in seawater as a feedstock has theadvantage of eliminating the load on freshwater resources for their cultivation.
The halophilic cyanobacterium Arthrospira (Spirulina) platensis is a potential
microalgal feedstock because it accumulates a large amount of glycogen andgrows faster than other microalgae. Moreover, A. platensis cultures have theadvantage of being resistant to bacterial contamination because they can growunder conditions of high pH and high salinity. Accordingly, A. platensis is
expected to be a promising supplier of carbohydrate in the form of glycogen. Also,
we developed direct ethanol production from microalgal carbohydrate by using
one of our core technologies, cell surface engineering of Saccharomyces
cerevisiae. In this symposium, we will introduce our project, development ofbioethanol production from marine microalgae, which is constructed from 1)
development of mass cultivation system of A. platensis under marine watercondition, 2) development of a novel genome modification technology for
microalgae, 3) system biology analysis of microalga, 4) improvement ofphotosynthetic capacity of microalgae through genetic engineering, 5)development of ethanol production system from microalgal glycogen by using cellsurface engineering.

Fri 26, OCT

4Aa06

SBJ/JST Joint Symposium II -Biorefinery-

Chlamydomonas reinhardtii - A powerful host for the production of
biofuels and high value products○Olaf KRUSE

Abstracts

Bielefeld University; Department of Biology/ Center for Biotechnology
Algae Biotechnology & Bioenergy GroupThe increasing interest on the application of phototrophic microorganisms as a
production host for sun energy based bio-products is reflected by a variety ofworldwide established collaborative research initiatives working in the field ofalgae biotechnology.
This presentation will summarize recent advances achieved in our group with themicroalga C. reinhardtii and includes new molecular engineering approaches formore efficient sun-to-biomass conversion efficiency and the establishment of amechanism for the production/secretion of a high value product. In addition it will
be demonstrated that C. reinhardtii with its cellulose-free cell wall has the abilityof cellulose degradation and assimilation for growth, a phenomenon which has
been previously only shown for heterotrophic bacteria and fungi. Phototrophic
microbes like C. reinhardtii may thus serve as bio-catalysts in cellulosic biofuel
production approaches.
Finally, the identification of a new microalgal candidate for an efficient bio-refinery
concept including rapid cell growth, high lipid content and good biogas production
yields will be presented.
[Keywords] Microalgae, C. reinhardtii, Biofuels, Cellulose, Light harvesting68

SBJ/JST Joint Symposium II -Biorefinery-

Alkenone biosynthesis by marine Haptophytes as biorefinery forrenewable energy production

Fri 26, OCT

4Aa07○Yoshihiro SHIRAIWA1,2
1

Faculty of Life & Environmental Sciences, University of Tsukuba, Japan
CREST, Japan Science and Technology Agency (JST), Japan

2[Keywords] Alkenone biosynthesis, Biofuel, Biorefinery,
Haptophyte algae, Lipid, Long-chain ketone, Metabolome

Coccolithophorid,

69

Abstracts

Marine microalgae have contributed to change in global environment by reducing
carbon dioxide concentration and increasing oxygen concentration. Such changes
have been the motive force for the evolution of microalgae during three billion
years. Coccolithophorids (Haptophytes), marine unicellular calcifying algae
producing calcium carbonate crystals as cell covering and lipid droplet as storage
compound, are known to be sources for lime stones and petroleum produced at
the Cretaceous era. According to literatures, the white cliff in Dover and petroleum
in the Middle East countries were produced from huge mass of inorganic andorganic cell components of such coccolithophorids, respectively. Coccolithophorids
are still producing a huge bloom even in the present ocean especially at highlatitude ocean such as the North Atlantic Ocean and the Bering Sea. In this talk, I
will focus on “the potential of haptophytes” for the reduction of atmospheric CO2
and especially the production of renewable energy source. Some of haptophyte
algae do not produce triglycerides but produce oil droplets such as long-chain
ketones, namely alkenones and alkens, which are associated with bio-oil
production, in the cells. These metabolites occupy ca. 30% of cell dry mass andare candidates for liquid and gaseous hydrocarbons as biofuels. Our researches
on alkenone producing haptophytes, their growth regulation, mechanism formetabolic analysis of alkenone biosynthesis and efficient production of usefulmetabolites and the development of lipid metabolomics will be introduced.

Fri 26, OCT

4Ap01

SBJ/JST Joint Symposium III -Biofuel Production by Microalgae-

Characterization of hydrocarbon biosynthesis and secretionmechanisms by the green microalga, Botryococcus braunii tocontrol biofuel production○Shigeru OKADA1,5, Ikuro ABE2,5, Tetsuko NOGUCHI3,5, Takeshi OHAMA4,5
1

Abstracts

Department of Aquatic Biosciences, Graduate School of Agricultural and Life
Sciences, The University of Tokyo
2
Graduate School of Pharmaceutical Sciences, The University of Tokyo
3
Department of Biological Sciences, Faculty of Science, Nara Women’s University
4
Department of Environmental Systems Engineering, Kochi University of Technology
5
CREST, Japan Science and Technology Agency (JST)The green microalga Botryococcus braunii produces unusually large amounts ofliquid hydrocarbons and secretes them outside of cells. One of the chemicalraces of this alga produces specific triterpenes, botryococcenes andmethylsqualenes, that are promising as an alternative fuel source since they arechemically very similar to petroleum. The genes coding for enzymes responsible
for the synthesis of these triterpenes, squalene synthase-like proteins (SSL)-1, -2,
and -3, were recently identified (Niehaus et al., 2011). The objective of ourresearch is to uncover “why and how” these triterpenes are produced andsecreted by this alga not only at the cellular but also the molecular level by four
strategies; 1) Characterization of the mechanism for gene expression related totriterpene biosynthesis in order to regulate hydrocarbon production, 2) Analysis of
the reaction mechanisms for SSL-1, -2, and -3 based on X-ray crystallography tomodify their functions for more effective hydrocarbon production, 3) Microscopic
observations of intercellular organelles in terms of hydrocarbon production andsecretion to understand the unique system of hydrocarbon secretion by the alga,
and 4) Establishment of techniques for transformation of the B. braunii in order to
identify functions of unknown genes or to generate new algal strains moresuitable for biofuel production. Information on expression patterns of the genes
concerned with hydrocarbon production during culture periods, the importantamino acid residues in SSL-1 that would recognize carbon chain length of thesubstrate, and the relation between hydrocarbon secretion and cell division cycles
were accumulated. Techniques that may enhance introduction of exogenous
genes were also developed.
T. D. Niehaus, S. Okada, T. P. Devarenne, D. S. Watt, V. Sviripa, and J. Chappell(2011) PNAS, 108: 12260-12265[Keywords] Microalgae, Botryococcus, Hydrocarbon, Biosynthesis, Transformation70

SBJ/JST Joint Symposium III -Biofuel Production by Microalgae-

Metabolic engineering of cyanobacteria for the light-powered
production of hydrogen from water

Fri 26, OCT

4Ap02○Matthias RÖGNER
Plant Biochemistry, Faculty of Biology & Biotechnology, Ruhr-University Bochum,
Bochum, Germany

Esper, B., Badura, A. & Rögner, M. (2006) Trends in Plant Sci. 11, 543-549
Bernat, G., Waschewski, N. & Rögner, M. (2009) Photosynth. Res. 99, 205-216
Waschewski, N., Bernat, G. & Rögner, M. (2010) in: “Biomass to Biofuels –Strategies for Global Industries” (Vertes, A., Qureshi, N., Yukawa, H.,
Blaschek, H.P. eds.) John Wiley & Sons, Chichester, UK, p. 387-401
Imashimizu, M., Bernát, G., Isato, K., Broekmans, M., Konno, H., Sunamura, E.-I.,
Rögner, M., Hisabori, T. (2011) J. Biol. Chem. 286, 26595-26602
Badura, A., Kothe, T., Schuhmann, W. & Rögner, M. (2011) Energy Environ. Sci. 4,3263-3274[Keywords] Biohydrogen production, Design cell, Cyanobacteria, Photosynthesis,
Biobattery, Photoelectrochemistry71

Abstracts

In order to produce hydrogen as potential future renewable energy source fromwater, we propose to engineer cyanobacterial photosynthesis towards increased
bioenergy instead of biomass production. Besides the implementation of a highly
active, oxygen tolerant hydrogenase from other organisms such as green algae(this requires a special strategy...), especially the photosynthetic electron
metabolism has to be engineered in many individual steps towards this goal. The
result of each single engineering step (such as antenna size reduction, partial
uncoupling of the thylakoid membrane, re-routing of electrons at the photosystem
1 acceptor site) has to be monitored by both functional (for instance spectroscopy)
as well as metabolic characterization on the whole cell level (for instance by an in
depth quantitative proteome, lipidome- and metabolome analysis). The directionof engineering is supported by studying protein-protein-interactions in isolated
model systems – for instance the affinity of Ferredoxin (Fd) for Ferredoxin-NADPOxidoreductase (FNR) vs. hydrogenase, which is a decisive step for re-routing
electrons from water for hydrogen production instead of CO2-fixation.
Performance of such engineered cells has to be optimized by improving
fermentation conditions and by an optimal photobioreactor design. Optimal culture
conditions can be found and kept constant for several months by using continuous
flow fermentation techniques which allow the systematic optimization of eachindividual parameter. Provided such systems are optimized both on the individual
cell level and on the systems level, a more than 100-fold increase of hydrogenproduction in comparison with the most productive natural systems existing todate can be estimated, which would be a promising basis for an economicallycompetitive H2-production.

Fri 26, OCT

4Ap03

SBJ/JST Joint Symposium III -Biofuel Production by Microalgae-

Selection strategies for the promising microalgal strains foralgae-fuel production○Hideaki MIYASHITA1, Norihide KURANO2
1

Abstracts

Department of Interdisciplinary Environment, Graduate School of Human andEnvironmental Studies, Kyoto University
2
Research laboratories, DENSO CORPORATION

Bio-fuel production using microalgae is a sustainable system based on oxygenic
photosynthesis using solar irradiation, CO2 and water. Therefore, the algae-fuelhas been actively investigated as the alternative source of energy against the oilcrisis in the future, as well as for a CO2 recovering process to prevent the green
house effects on the Earth. However, only a few large-scale systems forcommercial production have been established, since the algae-fuel is uneconomic
since the production cost is very high at present. Interdisciplinary researches
including phycology, physiology, molecular biology, chemistry, chemicalengineering and so on are required to cut down the production cost. Strain
selection of microalgae is one of the effective strategies for the cost reduction.
Even though more than 35,000 species of algae have already been described,
genera that have been evaluated for large-scale production are not so many evennow. Moreover, the number of species described until recently was reported to bejust several % of estimated algal species on the Earth. Therefore, continuous
efforts to isolate new algae from natural environments as well as the systematic
selection of promising oil-producers are extremely important. We have continued
to isolate and evaluate microalgae which are suitable for large-scale fuelproduction using open pond system, and successfully obtained several strains
which are efficient for fuel production. On the way of the selection, we set
microalgal abilities required for open pond system. Those are high biomass
productivity at low pH under high (>35˚C) or low (<15˚C) temperatures, and oil
productivity. In this presentation, we would like to introduce our strategies foralgae isolation and selection for fuel production using open pond system, in
addition to the growth characteristics of selected microalgae.
[Keywords] Microalgae-fuel, Strain selection strategy, Open pond72

SBJ/JST Joint Symposium III -Biofuel Production by Microalgae-

Elucidation of novel triterpene pathways in Botryococcus andengineering plants for high-value oil production

Fri 26, OCT

4Ap04

Tom D. NIEHAUS1, Scott KINISON1, Shigeru OKADA 2, Shuiqin WU1,
Timothy P. DEVARENNE3, David S. WATT4, Vitaliy SVIRIPA4, ○Joe CHAPPELL1
1

Plant Biology Program, University of Kentucky, Lexington, KY, 40546-0312, USA
Department of Aquatic Biosciences, University of Tokyo, Tokyo 113-8657, Japan3
Department of Biochemistry and Biophysics, Texas A&M University, College Station,
TX, 77843-2128, USA
4
Department of Cellular and Molecular Biochemistry, University of Kentucky, Lexington,
KY, 40536, USA
2[Keywords] Triterpenes, Botryococcene, Metabolic engineering

73

Abstracts

Assuming biofuels generated via the fermentation of sugars derived from
cellulosic and non-cellulosic constituents of biofuels crops will provide asubstantial contribution to our future energy needs, augmenting and amending
the productivity of these biofuel crops is now a major research thrust worldwide.
One way of enhancing these biofuels crops will be to engineer them for valueadded components such as oils that can be used for efficient fuel production andthe manufacturing of other high-value products currently derived from petroleum
oils. Towards this end, we have developed an engineering strategy for optimized
production of long, branched-chain hydrocarbon biosynthesis in plants. Branchedchain hydrocarbons, like methylated triterpenes, are readily cracked into paraffins
and naphthenes that can either be distilled to combustible fuels (gasoline, jet anddiesel), or can be used directly for the synthesis of plastics, nylons, paints andother oil-derived products manufactured by diverse chemical industries. Our
working hypothesis has been that success in generating high level production
platforms for triterpene oils in plants can be accomplished by targeting thismetabolism to unique compartments within the cell, thus eliminating the regulatory
mechanisms that normally operate to control this metabolism occurring in thecytoplasm, and providing a means for the direct channeling of photosynthetically
fixed CO2 to the biosynthesis of novel, value-added products. Preliminary
experiments suggests that we have been able to create plants yielding up to 1%of their dry weight as triterpenes. These results have important ramifications forfurthering our understanding of basic metabolism in plants, as well as thedevelopment of novel chemical production platforms in plants.

Fri 26, OCT

4Ap05

SBJ/JST Joint Symposium III -Biofuel Production by Microalgae-

The CyanofactoryTM~ a novel cyanobacterial bioprocess based on the synthetic biology
concept ~○Koji SODE1,2
1

Abstracts

Tokyo University of Agriculture & Technology, 2-24-16 Naka-cho, Koganei-shi, Tokyo,
184-8588, Japan
2
Japan Science and Technology Agency, CREST, 2-24-16 Naka-cho, Koganei-shi,
Tokyo, 184-8588, Japan

Our research group is engaged in the development of a novel bioprocess basedon the synthetic biology concept, for bioenergy production, designated as the“Cyanofactory TM”. The Cyanofactory TM is composed of 1) synthetic marine
cyanobacterial host strains, 2) synthetic operons for the production of biofuelrelated compounds, and 3) the employment of ionic liquids for downstream
processing.
“Synthetic marine cyanobacterial host strains” are the engineered cyanobacteria
whose functions are highly controlled by the artificial signal transduction system,
which is composed of the light sensors / histidine kinases, the response regulator
and the riboregulator / riboswitch systems. This system counts light stimulation
frequency and controls cell growth, biofuel-related compound production, selfaggregation and auto-lysis by regulation of the gene expression necessary foreach process. Finally, the produced biofuel-related compounds are extracted byionic liquids (ILs), which is recognized as a “Green solvents”. The combination ofdownstream processes employing ILs realizes the sustainable production of
biofuel-related compounds based on synthetic cyanobacterial processes withminimal energy and waste.
[Keywords] Cyanofactory, Cyanobacteria, Synthetic biology, Ionic liquid, Signaltransduction

74

SBJ/JST Joint Symposium IV -System & Synthetic Biotechnology-

Bio-refinery from microalgae through fermentation and metabolicengineering

Fri 26, OCT

4Ap06○Tomohisa HASUNUMA 1,21

Organization of Advanced Science and Technology, Kobe University
PRESTO, Japan Science and Technology Agency (JST)2[Keywords] Microalgae, Metabolic engineering, Bio-refinery, Bio-ethanol

75

Abstracts

Bio-refinery, the utilization of biomass as the starting material for the productionof fuels and chemicals, has received considerable attention in recent years.
Environmental concerns and the depletion of oil reserves have resulted ingovernmental actions and incentives to promote research on environmentallybenign and sustainable bio-fuels such as bio-ethanol. Cyanobacteria has
emerged as a feedstock for producing bio-fuels since many microalgal species
have the ability to assimilate CO2 and accumulate large amounts of carbohydrate
in the form of starch or glycogen in cells.
The halophilic cyanobacterium Arthrospira (Spirulina) platensis was investigatedfor its feasibility to serve as feedstock for fermentative production of bio-ethanol.
The use of salt-tolerant microalgae grown in seawater as a feedstock has theadvantage of eliminating the load on freshwater resources for their cultivation.
Moreover, A. platensis cultures have the advantage of being resistant to bacterial
contamination because they can grow under conditions of high pH and highsalinity. With optimal control of both light intensity and nitrate supply, glycogenproduction of A. platensis reached nearly 1.03 g/L (a glycogen productivity of 0.29
g/L/d), which is, to the best of our knowledge, the highest α-polyglucan production
performance ever reported in microalgae. In the present study, glycogenproduced by A. platensis was directly converted to ethanol by a recombinant
Saccharomyces cerevisiae strain expressing both Streptococcus bovis α-amylase
and Rhizopus oryzae glucoamylase with more than 80% of theoretical yield.
For the enhancement of glycogen productivity in cyanobacteria, glycogenmetabolism would be preferred to be modified through metabolic engineering.
Glycogen is significantly accumulated in the cyanobacterial cell by the depletionof nitrate. We have developed an analytical system of dynamic metabolic profilingin cyanobacteria with metabolomics and in vivo 13C-labeling technique. In thepresent study, metabolic intermediates involved in the Calvin cycle, glycolysis,
TCA cycle, amino acid biosynthesis have determined during the perturbation bynitrate depletion. Based on the metabolic profiling system, putative mechanism of
glycogen biosynthesis was successfully assumed under the nitrate depleted
condition.

Fri 26, OCT

4Ap07

SBJ/JST Joint Symposium IV -System & Synthetic Biotechnology-

Synthetic biology approaches to produce C3-C5 alcohols frommicroorganisms○Shota ATSUMI

Abstracts

Department of Chemistry, University of California Davis

Biofuels synthesized from renewable resources are of increasing interest becauseof global energy and environmental problems. Compared to the traditional biofuel,ethanol, higher alcohols such as isobutanol and 1-butanol offer advantages suchas higher energy density, lower hygroscopicity, lower vapor pressure, andcompatibility with existing transportation infrastructure. Some Clostridia species
are known to naturally produce isopropanol and 1-butanol. However, these fuels
are not synthesized economically using native organisms. Additionally, other C3C5 alcohols are not produced in large quantities by natural microorganisms.
Synthetic biology offers an alternative approach in which synthetic pathways areengineered into user-friendly hosts for the production of these fuel molecules. We
built up unnatural synthetic pathways to produce higher-order alcohols. Moreover,
we improved the productivity by combining gene deletion and overexpression
techniques. Our demonstration shows that the strategy enables the exploration of
biofuels beyond those naturally accumulated to high quantities in microbial
fermentation.
[Keywords] Isobutanol, Biofuel, Synthetic biology76

SBJ/JST Joint Symposium IV -System & Synthetic Biotechnology-

Synthetic metabolic engineering –A novel, simple technology fordesigning a chimeric metabolic pathway–

Fri 26, OCT

4Ap08○Kohsuke HONDA1,2, Xiaoting YE1, Kenji OKANO1, Hisao OHTAKE1
1
2

Department of Biotechnology, Graduate School of Engineering, Osaka University
PRESTO, Japan Science and Technology Agency (JST)[Keywords] Synthetic metabolic engineering, Thermophile, Embdem-Meyerhof
pathway

77

Abstracts

A novel, simple technology, designated as “synthetic metabolic engineering”, forfreely designing a chimeric metabolic pathway for bio-based production was
proposed. In this technology, genes encoding thermophilic enzymes were
overexpressed in mesophilic host. The excellent thermal stability of thermophilic
enzymes allows us to obtain whole-cell biocatalysts as highly selective as purified
enzymes by simply heating the resulting recombinant mesophiles. The in vitro
synthetic metabolic pathways can be constructed by rational combination of these
biocatalytic modules since this approach is applicable to all thermophilic enzymes
as long as they can be functionally expressed in heterologous mesophiles. Owing
to their independence from biological activity of living cells, synthetic pathways nolonger play the physiological roles of natural metabolism involving an energy
generation (catabolism) and energy consuming synthesis of biomolecules(anabolism), and thus a bespoke pathway specialized to chemical conversion canbe constructed. In this work, we constructed a chimeric glycolytic pathway, inwhich the consumption and regeneration rates of ATP and ADP are balanced, by
combinatorially using 7 glycolytic enzymes of Thermus thermophilus, the cofactorindependent phosphoglycerate mutase of Pyrococcus horikoshii, and the nonphosphorylating glyceraldehyde 3-phosphate dehydrogenase of Thermococcus
kodakarensis. By coupling this pathway with a thermophilic malate/lactate
dehydrogenase, a stoichiometric amount of lactate could be produced fromglucose with an overall ATP turnover number of 32.

Fri 26, OCT

4Ap09

SBJ/JST Joint Symposium IV -System & Synthetic Biotechnology-

Production of BioIsopreneTM Monomer○Derek H. WELLS

Abstracts

DuPont Industrial Biosciences

DuPont Industrial Biosciences and The Goodyear Tire & Rubber Company aredeveloping a bio-based alternative to petroleum-derived isoprene, a key chemical
required to produce a diverse range of products including specialty elastomers,
motor mounts and fittings, rubber bands, golf balls, shoes and tires. The annual
world market potential for polymer grade isoprene is approximately 2 billion
dollars. Biological production of isoprene (BioIsopreneTM monomer) is a nonpetrochemical process that directly addresses the lack of supply and priceinstability of this valuable commodity.
The biosynthetic production of isoprene presents significant metabolic engineering
challenges, such as pathway enzyme regulation, carbon to product channeling,redox and energy balancing, and selection of appropriate enzymes. High titers of
BioIsopreneTM monomer have been achieved by the introduction of an isoprenoid
pathway and an engineered isoprene synthase into Escherichia coli. Analysis has
demonstrated that the BioIsoprene™ product is >99% pure prior to recovery andpurification. Pre-pilot work has led to the manufacture of prototype passenger car
tires demonstrating the functional process from start to finish.
[Keywords] DuPont, Isoprene, Isoprenoid, Metabolic engineering, Escherichia coli78SBJ/JST Joint Symposium IV -System & Synthetic Biotechnology-

Evolutionary pathway engineering: toward the fast-track
construction of long-step, but high-fidelity pathway for non-naturalcompounds

Fri 26, OCT

4Ap10○Daisuke UMENO1,2, Maiko FURUBAYASHI1, Kyoichi SAITO1
1

Department of Applied Chemistry and Biotechnology, Graduate School ofEngineering, Chiba University
2
PRESTO, Japan Science and Technology Agency (JST)[Keywords] Directed evolution, Secondary
regulatory networks, Carotenoids, Isoprenoids

metabolism,

Specificity,

Gene

79

Abstracts

Rapid expansion of available biosynthetic gene is enabling synthetic biologists tostart constructing various pathways for non-natural compounds. However,targeted synthesis of non-natural compounds remains a challenge for two
reasons. First, non-natural pathways fatally contain non-existing biochemical
steps, and we have to create all and each of them. Second, due to the promiscuity
or imperfectness of biosynthetic parts, simple assemblage of biosynthetic genes
generates hyper-branched pathway that divert precursors into numerous
unwanted compounds. Here, we demonstrate the construction of long-step
pathway to a series of carotenoids and steroids with non-natural backbones
through the iterative rounds of directed evolution to create novel biochemical
steps, generates diversity in substrate/ product specificity, followed by thestrategic assemblage of the variant enzymes. We show that this way we canconstruct highly-specific pathways toward a targeted molecule out of thepromiscuous enzymes.

Index

Authors IndexA

ABE Ikuro
ARA Takeshi
ARITA Masanori
ATOMI Haruyuki
ATSUMI Shota

B

BUECHS JochenBÜHLER BrunoBÜHLER KatjaCCARSTENSEN Frederike
CHANG Jo-Shu
CHAPPELL Joe
CHEN Chun-Yen
CHEN Yong
CHOI Yong-Eui
CHOU Yi-Ting

D

DE MARIA Pablo Domínguez
DEVARENNE Timothy P.

F

FUKUSAKI Eiichiro
FURUBAYASHI Maiko
FURUSAWA Chikara

G

GRANDE Philipp

HAN Jung-Yeon
HASUNUMA Tomohisa
HATANAKA Haruyo
HIRASAWA Takashi
HONDA Kohsuke

I

IKEZAWA Nobuhiro
IMANAKA Tadayuki
ISHIKAWA Shu
ITAYA Mitsuhiro

2Ap06
2Ap01
2Ap01

2Ap06
2Ap09
4Ap04
2Ap09
3Fa04
2Fa07
3Fa05

2Ap06
4Ap04

3Ap08
4Ap10
2Ap10

J

JAEGER Gernot
JIN Yong-Su
JO Jung-Hyun
JULSING Mattijs
JUNG Sang-Min

K

KAMIYA Noriho
KATAYAMA Arata
KATO Tatsuya
KAWANO Shigeyuki
KIM Byung-Gee
KIM Seung Wook
KINISON Scott
KLEMENT Tobias
KONDO Akihiko
KRUSE Olaf
KUNZE Martin
KURANO Norihide

L

LEE Choul-Gyun
LEE Ja Hyun
LEE Nahum
LEE Sang Jun
LEE SangHyuk
LIAO James C.

M
2Ap06

2Fa07
4Ap06
3Ap05
2Ap10
4Ap08

2Fa04
4Aa01
2Ap03
2Fa03

MAEDA Tomohide
MATSUNAGA Tadashi
MIKI Kunio
MITSUDA Nobutaka
MIYASHITA Hideaki
MURANAKA Toshiya

N

NARANGAJAVANA Jarunya
NARUSE Kazuya
NGUYEN Don Trinh
NIEHAUS Tom D.
NISHIYAMA Kazue
NOGUCHI Tetsuko

2Ap06
3Ap06
3Ap06
2Ap01
3Ap06

3Fa06
3Fa01
3Ap07
4Aa03
2Ap07
3Ap10
4Ap04
2Ap06
4Aa05
4Aa06
2Ap06
4Ap03

4Aa02
3Ap10
2Ap07
3Ap10
2Ap07
2Ap02

3Ap05
3Ap01
4Aa01
2Fa02
4Ap03
2Fa08

2Fa05
3Ap07
2Fa04
4Ap04
3Ap05
4Ap01

83

Index

H

4Ap01
2Fa06
2Ap08
4Aa01
4Ap07

O

OGASAWARA Naotake
OGINO Hiroyasu
OHAMA Takeshi
OHME-TAKAGI Masaru
OHTAKE Hisao
OKADA Shigeru
OKADA Shigeru
OKAMOTO Masahiro
OKANO Kenji
ONO Naoaki
ONUMA Chumsakul
OTA Shuhei

2Ap03
3Fa02
4Ap01
2Fa02
4Ap08
4Ap01
4Ap04
2Ap05
4Ap08
2Ap10
2Ap03
4Aa03

SONG ChangHyun
SPIESS Antje
SUZUKI Hideyuki
SVIRIPA Vitaliy

T

TANG Jingchun
THAPA Laxmi Prasad
TOYA Yoshihiro
TREDICI Mario R
TRENT Jonathan D.

U

UMENO Daisuke

P

PARK Enoch Y.
PARK Sunghoon
PARK Yong-Cheol
PRASETYO Joni
PULZ Otto

Q

QU VinceR

RO Dae-Kyun
RÖGNER Matthias

Index

S

SAITO Kyoichi
SAKURAI Nozomu
SCHMID Andreas
SCHUBERT Ingo
SEKI Hikaru
SEO Jin-Ho
SHIBATA Daisuke
SHIMIZU Hiroshi
SHIRAIWA Yoshihiro
SODE Koji

84

3Ap07
3Ap09
3Ap06
3Ap07
4Aa04

2Fa04

2Fa04
4Ap02

W

WANG Hui-Min
WATANABE Koichi
WATT David S.
WELLS Derek H.
WESSLING Matthias
WEWETZER Sandra
WIJFFELS René H.
WU Jinglan
WU Pei-Fang
WU Shuiqin

Y
4Ap10
2Fa06
2Ap01
2Fa01
2Fa08
3Ap06
2Fa06
2Ap10
4Aa07
4Ap05

YASUE Yuri
YE Xiaoting
YING Hanjie
YONG Yang-Chun
YOO Hah Young
YOO Tae Hyeon
YOSHIDA Ken-ichi
YOSHIKAWA KatsunoriZ

ZHONG Jian-Jiang

2Ap07
2Ap06
2Fa06
4Ap04

3Fa01
3Ap10
2Ap10
3Ap03
3Ap02

4Ap10

3Fa05
4Aa03
4Ap04
4Ap09
2Ap06
2Ap06
3Ap04
3Fa04
3Fa05
4Ap04

3Ap05
4Ap08
3Fa04
2Ap04
3Ap10
3Fa03
2Ap03
2Ap10

2Ap04

Keywords Index
A

Adsorption immobilization
3Fa04
Algae
3Ap01, 3Ap02
Alginate degradation
3Fa01
Alkenone biosynthesis
4Aa07
Alzheimer’s disease
2Ap03
Amino acid analogues
3Fa03
Aminoacyl-tRNA synthetase
3Fa03
Anti-bacterial
3Fa05
Anti-cancer
3Fa05
Anti-oxidant
3Fa05
Anti-pigmentation
3Fa05
Aquaculture
3Ap02
Aquatic microorganisms
3Ap01
Archaea
4Aa01
Artificial chromosomes
2Fa01
Astaxanthin
4Aa03
Asteraceae
2Fa04
Asymmetric epoxidation
2Ap01

B

C. reinhardtii
4Aa06
Carbohydrate
2Ap09
Carotenoids
4Ap10
Cassava
2Fa05
Cell retention
2Ap06
Cellulase
3Ap07
Cellulose
4Aa06
Cellulosic biomass
3Ap06
Centromere maintenance
2Fa01
Cephalosporin C
3Ap10
Chimeric repressor
2Fa02
Chlorella
4Aa03
Chromosome biology
2Fa01
Coccolithophorid
4Aa07
Cofactor manipulation
3Fa04
Composting
3Fa01
Continuous culture
2Ap06
Cross-linking of biomaterials
2Ap07
Cyanobacteria
4Ap02, 4Ap05
Cyanofactory
4Ap05
Cytochrome P450
2Fa04

D

Dammarane-type ginsenoside
Database
Denaturing gradient gelelectrophoresis
Design cell
Directed evolution
3D reconstruction
DuPont

2Fa07
2Ap08
3Fa01
4Ap02
4Ap10
4Aa03
4Ap09

E

Embdem-Meyerhof pathway
4Ap08
Embryological engineering
2Ap05
Energy crop
2Fa05
Energy return on investment
3Ap03
Enzyme immobilization
3Ap10
Escherichia coli
4Ap09
Evolution engineering
2Ap10
Evolutional molecular engineering
2Ap05

85

Index

Bacillus subtilis
2Ap03
Biobattery
4Ap02
Bio-butanol
3Fa04
Bioconversion
2Ap03
Biodiesel
3Ap10
Bioelectricity
2Ap04
Bioethanol
3Ap05, 3Ap06, 3Ap07, 3Ap10
Bio-ethanol
4Aa05, 4Ap06
Biofuel
3Ap01, 4Aa07, 4Ap07
Biofuels
2Ap09, 3Ap02, 4Aa06
Biohydrogen production
4Ap02
Bioinformatics
2Fa06
Biomedicine
2Ap04
Bioprocess engineering
2Ap04
Biorefinery
2Ap09, 3Ap04, 3Ap07, 4Aa07
Bio-refinery
4Aa05, 4Ap06
Biorefining
2Ap02
Biosynthesis
4Ap01
Botryococcene
4Ap04
Botryococcus
4Ap01

C

F

Flue gas
Functional genomics

G

GC/MS
Gene expression
Gene regulation
Gene regulatory networks
GenomeGenome design
Genome scale model
GingerGlycerol
Glycerol channel
Green wall panel (GWP)

H

Haematococcus
Haptophyte algae
Hydrocarbon
3-Hydroxypropionic acid
Hyperthermophile

I

3Ap08
2Fa03
2Fa02
4Ap10
2Fa05
2Fa03
2Ap10
3Fa05
3Ap09
3Ap05
3Ap03

4Aa03
4Aa07
4Ap01
3Ap09
4Aa01

Mass spectrometry
2Fa06
Membranes
2Ap06
Metabolic engineering 2Fa08, 2Ap02,
2Ap10, 3Ap09, 4Ap04,
4Ap06, 4Ap09
Metabolic flux analysis
2Ap10
Metabolic pathways
3Ap01
Metabolic profiling
3Ap08
Metabolism
4Aa01
Metabolome
3Ap08, 4Aa07
Metabolome analysis
2Fa06
Metabolomics
2Ap08, 3Ap08
Micro fluid engineering
2Ap05
Microalgae
2Ap09, 3Ap03, 3Ap04,
4Aa04, 4Aa05, 4Aa06,
4Ap01, 4Ap06
Microalgae-fuel
4Ap03
Microalgal biodiesel
4Aa02
Microreactors
2Ap01
2Ap09
Mitigation of CO2 emissions
Molecular breeding
2Fa05
Multi variate analysis
3Ap08
Multiomics analysis
2Ap10
Multistep reactions
2Ap01

2Ap03
2Ap06
2Fa01
4Ap05
3Fa06
4Ap07
4Ap09
4Ap09
4Ap10
3Fa04

N

Light harvesting
4Aa06
Lignocellulose
2Ap06
Lignocellulosic biomass
3Fa06
Lipase
3Fa02
Lipid
2Ap09, 4Aa07
Long-chain ketone
4Aa07

P

InositolIntegrated process
Interphase chromatin dynamics
Ionic liquid
Ionic liquids
Isobutanol
Isoprene
Isoprenoid
Isoprenoids
ISPR

Index

2Ap09
2Fa05

L

M

Marine microalgae
Mass cultivation
Mass spectra

86

4Aa02
4Aa04
2Ap08

NAD kinase
Natural rubber
Next generation sequencing

O

Ocean culture
Omics
Open pond
Organic solvent
ortho-Dihydroxylation

3Ap05
2Fa04
2Fa04

4Aa02
3Ap01
4Ap03
3Fa02
2Ap07

P450
2Fa08
Panax ginseng
2Fa07
Paper sludge
3Ap07
Photobioreactor 3Ap02, 3Ap03, 4Aa02
Photobioreactors
4Aa04
Photoelectrochemistry
4Ap02
Photosynthesis
2Ap02, 4Ap02
Photosynthesis and photovoltaics
3Ap03
Productivity
2Ap011,3-Propanediol
Protease
ProteinProtein engineering

Q

Quinone profile method
Quorum sensing

3Ap09
3Fa02
2Ap09
3Fa03

3Fa01
2Ap04

R

Recombinant Saccharomyces
cerevisiae
Regulation
Ribosomal RNA

S

Sulfur-deficient
4Aa03
Synthetic biology
2Fa08, 2Ap02, 4Ap05, 4Ap07
Synthetic centromeres
2Fa01
Synthetic genetic circuit
2Ap05
Synthetic isozyme system
3Ap06
Synthetic metabolic engineering
2Ap05, 4Ap08

T
3Ap06
4Aa01
2Fa03

Saccharification
3Ap07
Saponin biosynthesis
2Fa07
Secondary metabolism
4Ap10
Sesquiterpene
2Fa04
Signal transduction
4Ap05
Signal transduction engineering 2Ap04
Solvent engineering
3Fa06
Specificity
4Ap10
Stability
3Fa02
Strain selection strategy
4Ap03
Streptomyces
2Ap07
Sugar transport
3Ap06

Tetracyclic triterpene
Thermophile
Tolerance
Transcription factor
Transcriptome
Transformation
Triterpenes
Triterpenoid
Tyrosinase

W

Wastewater treatment
Whole cell biocatalysis
Whole genome cloning

X

Xylose

2Fa07
4Ap08
3Fa02
2Fa02
2Fa06
4Ap01
4Ap04
2Fa08
2Ap07

3Ap02
2Ap01
2Fa03

3Ap05

Index

87

Journal of

Bioscience andBioengineering
114
1

NCE A
ND

JOURN
AL OF
BIOSC
IE

FOUNDED 1923

http://w

ww .sbj.or.jpPublished
The Socie Monthly byty for
Biotechnology,
Japan

July 20

12 Vo
lume 114 Issu
e1

RING

INEE

BIOEN
G

Journal of

12

JULY 20e 114

V olumBioscie
Bioeng nce andineeringIssue
1
P ages
1–
122

IER

ELSEV

http://
www.elsevier
Print ISSN 1389 .com/locate/jbioscOnline-1723
IS
CODEN: SN 1347-442
1
JBBIF6

Published by the Society for Biotechnology, Japan/ Distributed outside Japan by Elsevier

http://www.elsevier.com/locate/jbiosc
Print ISSN 1389-1723 Online ISSN 1347-4421 CODEN: JBBIF6

The90 th Anniversary MeetingThe

90 th Anniversary Meeting

The Society for Biotechnology, Japan
International Symposium on

Program & Abstracts

Program & Abstracts

October 24-26, 2012
Kobe International Conference Center

The Society for Biotechnology, Japan

Kobe, Japan

Organized by
The Society for Biotechnology, Japan (SBJ)
Japan Science and Technology Agency (JST)Supported by
Kobe Convention & Visitors Association
Nakauchi Tsutomu Convention Shinko Zaidan Foundation
Suntory Institute for Bioorganic Research