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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| Host Researcher |
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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| Foreign Research Fellow |
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 |
Granted (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 | 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 |
In this year we have mainly worked with the alanine:glyoxylate aminotransferase, because we discovered some intrinsically disordered regions on the protein surface. In detail we have performed several investigation on these regions from a bioinformatic (molecular dynamics), biochemical (library screening and production and mutants purification as well) and cellular point of view. In the meantime we are also working in the preparation of a new system to study enzymes behavior, generating enzymatically active protocells using the liquid-liquid phase separation.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The paper on the disordered regions of AGT is ready for submission and the other one of the enzymatically active protocells as well. We are just working on the final details before submission.
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| Strategy for Future Research Activity |
The project about DPAT did not give the expected results, we are still optimizing the assay and the engineering part of the protein. We are also currently working with the enzymatically active protocells , in detail we are studying the effects of enzymatic activity on the protocells environment.
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Report
(2 results)
Research Products
(2 results)
