Identification of the genes encoding biomass-degrading enzymes by microbial screening based on microdroplet deformability
| Project/Area Number |
18K05330
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 37020:Chemistry and chemical methodology of biomolecules-related
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Iizuka Ryo 東京大学, 大学院理学系研究科(理学部), 助教 (90541954)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | 液滴 / マイクロ流体デバイス / 酵素 / スクリーニング |
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| Outline of Final Research Achievements |
Environmental microbes secrete enzymes capable of degrading macromolecules, such as agarose, into smaller molecules for cell growth. These enzymes can benefit the effective use of renewable natural resources such as biomass. However, most microbial cells remain unculturable and thus are inaccessible by culture-based methods. We then developed a culture-independent method for screening microbial cells that secrete polysaccharide hydrogel-degrading enzymes using deformability-based microfluidic microdroplet sorting. In this method, microbial cells are encapsulated as single cells in water-in-oil (W/O) microdroplets with hydrogel, whose shape becomes deformable as the hydrogel is progressively degraded into smaller molecules. Screening is achieved using a microfluidic device that passively sorts the deformed W/O microdroplets. Using this method, we successfully isolated single bacterial cells that hydrolyze agarose from seawater to identify agarase genes.
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| Academic Significance and Societal Importance of the Research Achievements |
世界で6番目の広さの海洋に囲まれる我が国では，安定的に大量供給が可能で食糧と競合しないリグノセルロース系バイオマス（セルロース，ヘミセルロース，リグニンなど）とともに，海藻バイオマス（アガロース，アルギン酸，フコイダンなど海藻多糖類）の有効利用が重要である．海藻多糖類はエタノールなどのエネルギー原料のみならず，付加価値の高い生理活性分子に変換可能である．しかし，海藻多糖類を効率よく分解できる酵素の報告は少ない．本研究で確立したスクリーニング法を駆使することで，優れた海藻多糖類分解酵素の取得が可能となる．
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Report
(5 results)
Research Products
(53 results)
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[Journal Article] Construction of integrated gene logic-chip2018
Author(s)
Takeya Masubuchi, Masayuki Endo, Ryo Iizuka, Ayaka Iguchi, Dong Hyun Yoon, Tetsushi Sekiguchi, Hao Qi, Ryosuke Iinuma, Yuya Miyazono, Shuichi Shoji, Takashi Funatsu, Hiroshi Sugiyama, Yoshie Harada, Takuya Ueda, Hisashi Tadakuma
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Journal Title
Nat. Nanotechnol.
Volume: 13
Issue: 10
Pages: 933-940
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Presentation] Feasibility study of the method to obtain peptide agonists for G protein-coupled receptors using water-in-oil microdroplets2019
Author(s)
Anna Matsueda, Takashi Sakurai, Ryo Iizuka, Yasuyuki Nakamura, Jun Ishi, Akihiro Kondo, Dong Hyun Yoon, Tetsushi Sekiguchi, Syuichi Syoji, Soichiro Tsuda, Takashi Funatsu
Organizer
第57回日本生物物理学会年会
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[Presentation] Droplet-based microfluidic screening for obtaining microbes producing macromolecule-degrading enzymes2018
Author(s)
Ryo Iizuka, Kai Saito, Eiji Shigihara, Wataru Kawakubo, Daiki Tanaka, Dong Hyun Yoon, Tetsushi Sekiguchi, Shuichi Shoji, Mei Ito, Yuji Hatada, Takashi Funatsu
Organizer
第56回日本生物物理学会年会
Related Report
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[Presentation] High-throughput in vitro selection method for obtaining peptide agonists of G protein-coupled receptors2018
Author(s)
Anna Matsueda, Takashi Sakurai, Ryo Iizuka, Yasuyuki Nakamura, Jun Ishii, Akihiko Kondo, Ayaka Iguchi, Dong Hyun Yoon, Tetsushi Sekiguchi, Shuichi Shoji, Yuu Fujimura, Jin Akagi, Masayuki Ishige, Takashi Funatsu
Organizer
第56回日本生物物理学会年会
Related Report
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