非天然型アミノ酸生産のためのPLP依存性アミノ基転移酵素の改変
| Project/Area Number |
19F19764
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| Research Category |
Grant-in-Aid for JSPS Fellows
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| Allocation Type | Single-year Grants |
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| Section | 外国 |
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| Review Section |
Basic Section 43020:Structural biochemistry-related
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| Research Institution | Okinawa Institute of Science and Technology Graduate University |
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Principal Investigator |
LAURINO Paola 沖縄科学技術大学院大学, タンパク質工学・進化ユニット, 准教授 (90812256)
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| Co-Investigator(Kenkyū-buntansha) |
DINDO MIRCO 沖縄科学技術大学院大学, タンパク質工学・進化ユニット, 外国人特別研究員
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| Project Period (FY) |
2019-11-08 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2021: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2020: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2019: ¥100,000 (Direct Cost: ¥100,000)
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| Keywords | Phase separation / enzymes / enzyme kinetic / protein engineering / protein plasticity / Disordered regions / AGT / protocells / Liquid phase separation / Biochemistry / enzyme / library / screening |
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| Outline of Research at the Start |
The DPAT enzyme will be fully characterised from structural and functional perspectives, in order to understand its unusual activity in detail. Laurino group already has an established collaboration with the Jackson Lab at ANU, Australia and they will help to solve the structure of this enzyme. Then, BLAST searches will help us to find proteins structurally related to DPAT, in order to identify possible homologs with the same or similar functions and to expand our knowledge of this interesting class of enzymes. Finally the recombinant DPAT will be engineered by rounds of directed evolution.
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| Outline of Annual Research Achievements |
・The production/generation of liquid-liquid phase separated membrane-less droplets with novel features as the ability to compartmentalize enzymes efficiently, high protein crowding (mM protein content) to study in detail the effects of crowding on proteins and especially on the enzymatic activity (using length scale of living systems micrometer scale as cells).
・We also demonstrated how the enzymatic activity in presence of crowding generated flow similar to the cytoplasmatic streaming of the cell. This work has been published by Nature Communications it is amongst the 2021 top 25 most downloaded Chemistry and Material Science Articles in 2021 published in Nature communication.
・In addition, we have discovered how protein plasticity shape the function and fitness. The work has been published in Protein Science.
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| Research Progress Status |
令和3年度が最終年度であるため、記入しない。
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| Strategy for Future Research Activity |
令和3年度が最終年度であるため、記入しない。
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Report
(3 results)
Research Products
(4 results)
