| Project/Area Number |
17K15248
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied microbiology
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| Research Institution | Toyama Prefectural University |
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Principal Investigator |
Toda Hiroshi 富山県立大学, 工学部, 講師 (60608321)
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2018: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 有機溶媒耐性菌 / Kocuria rhizophila / キラル化合物 / シャトルベクター / 有機溶媒耐性微生物 / 代謝工学 / 物質生産 / ゲノム編集 |
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| Outline of Final Research Achievements |
1) We constructed novel E. coli-Kocuria shuttle vector pKITE301 and its derivatives using cryptic pKPAL3 plasmid from K. palustris IPUFS-1. Various enzymes including styrene monooxygenase, alcohol dehydrogenase, and flavonoid methyltransferase were expressed in K. rhizophila DC2201 using pKITE301 for producing chiral compounds and methylated flavonoids.
2) K. rhizophila DC2201 was cultured in the presence of an organic solvent, and the transcribed mRNA was comprehensively analyzed by next-generation sequencing. When K. rhizophila DC2201 cells were cultured in the presence of organic solvent, transcription level of several genes coding response regulator and transporter were markedly increased. Additionally, transcription level of several glycosyl transferases and mannose biosynthesis genes also increased. It was suggested that these genes were involved in synthesis of extracellular polysaccharide to improve organic solvent tolerance of Kocuria cells.
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| Academic Significance and Societal Importance of the Research Achievements |
有用化合物生産の宿主として有望な有機溶媒耐性菌K. rhizophila DC2201で利用な新規シャトルベクターを開発し、様々なバイオプロセスへの応用の可能性が見出された。特に細胞毒性が強い各種有機化合物の生産に応用可能なことから、医薬品中間体等の効率的な生産への応用が期待される。次世代シーケンスを利用したRNA転写解析を行った結果、K. rhizophila DC2201を有機溶媒に曝露した時に複数の糖転移酵素やマンノース生合成酵素の転写量が増加した。これらは細胞外多糖の合成に関与すると予測されることから、これらの機構を利用した有機溶媒耐性宿主の創製に応用可能であると期待される。
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