2023 Fiscal Year Research-status Report
Data-driven discovery of novel terpene synthases and its utilization for efficient production of structurally divergent compounds through a light control system in bacteria
| Project/Area Number |
23K13898
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Vo NgocQuynhNhu 国立研究開発法人理化学研究所, 環境資源科学研究センター, 特別研究員 (60888543)
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| Project Period (FY) |
2023-04-01 – 2026-03-31
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| Keywords | Terpene synthases / Drimenol / Sesquiterpenes / Marine bacteria / Gene clusters / Blue-light switch / Metabolic engineering |
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| Outline of Annual Research Achievements |
The goal of this study is to understand the diversity and biosynthetic potential of terpenoids in bacteria, especially marine bacteria. To achieve this goal, during the FY2023, as originally planned, I have investigated and found a number of promising bacterial terpene synthase genes, using our optimized genomic data mining method. I have characterized several terpene synthase enzymes by in vitro enzymatic assays and gas chromatography-mass spectrometry, and discovered novel enzymatic activities.
Moreover, I have successfully found the biosynthetic genes clustered with drimenol synthases encoded in marine bacterial genomes for drimenol synthesis, which are uniquely conserved. I have also verified the drimenol production in marine bacteria and suggested the evolutionary features of the drimenol gene cluster in marine bacteria. We have summarized these findings in a scientific paper and we will submit it for publication.
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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
As stated in the "Summary of Research Achievements", I have investigated and found more unknown terpene synthase genes of bacterial origin by genomic data mining and biochemical validation. In addition, I have elucidated the existence of core biosynthetic genes involved in drimenol biosynthesis and their distribution across marine bacterial genomes, verifying the accumulation of drimenol, a highly active sesquiterpene, in bacteria (manuscript in preparation). With those achievements, the status to date of my research project is progressing smoothly, which is approaching to the goal of advancing our understanding of the diversity and biosynthetic origin of terpenoids derived from bacteria including marine bacteria.
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| Strategy for Future Research Activity |
As for future work, I will further perform functional analysis using in vitro enzymatic assays and gas chromatography-mass spectrometry of promising candidate terpene synthases harvested from data mining approach to identify more novel enzymes with unique structures and reaction mechanisms that can make novel active terpenoids. In addition, I will elucidate biosynthetic genes, with a focus on prenyltransferases and monooxygenases, clustered with those characterized terpene synthases to make structurally divergent compounds. At the same time, I will construct and characterize the new bacterial blue-light switch platform to be ready for the introduction of novel bacterial terpene synthase genes and their clustered genes to enhance the production of target compounds.
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| Causes of Carryover |
During FY2023, I have applied new genomic data mining strategies that led me to harvest more unknown promising terpene synthase genes hidden in the bacterial genomes. I have successfully identified several novel enzymatic activities, so I have characterized those new enzymes in detail and prepared for the manuscript for publication. Therefore, I have decided to analyze the functions of other candidate genes and construct the blue-light switch system in the next FY2024. For these reasons, I have the remaining budget to be used for the next fiscal year. I plan to combine the "Amount to be used next fiscal year" with the grant requested for the FY2024 to purchase reagents and synthetic genes for characterizing other candidate genes in detail and performing blue-light experiments.
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Research Products
(2 results)
