2019 Fiscal Year Final Research Report

Elucidation of an ionic-liquid resistance mechanism of an ionic-liquid resistant bacterium and development of a novel microbial platform

Research Project

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Project/Area Number	17K07736
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Applied microbiology
Research Institution	Kindai University
Principal Investigator	Kurata Atsushi 近畿大学, 農学部, 准教授 (10416000)
Project Period (FY)	2017-04-01 – 2020-03-31
Keywords	イオン液体 / Bacillus 属細菌 / イオン液体耐性細菌 / 多剤排出トランスポーター / 浸透圧ストレス / ゲノム解析 / イオン液体耐性酵素 / RNA-seq
Outline of Final Research Achievements	Ionic liquids have physical properties for use as reaction solvents. To solve the problems of growth inhibition of microorganisms by ionic liquids, we isolated the ionic liquid-tolerant Bacillus sp. CMW1. The first objective of this study is to identify the gene which are responsible for ionic-liquid tolerant. The second objective is to develop the novel bacterial host which exhibits tolerance to ionic liquids. Consequently, I identified MFS transporter gene from strain CMW1 and showed that the gene is responsible for an ionic-liquid tolerant. The gene of CMW1 was transformed into Brevibacillus sp. bacterium and the recombinant strain could grow in the presence of an ionic liquid. Using tetracycline, pHY300PLK was successfully electroporated into strain CMW1 with the efficiency. I obtained the mutants which were able to grow in the presence of hydrophobic ionic liquids. The results will provide a basis for the future work in biocatalysis with various ionic liquids in industry.
Free Research Field	応用微生物学
Academic Significance and Societal Importance of the Research Achievements	イオン液体耐性宿主を開発できれば、イオン液体存在下でイオン液体耐性酵素を菌体外に生産できる。イオン液体で難分解性バイオマスを溶解できるため、イオン液体耐性酵素により、溶解したバイオマスを分解できる。イオン液体存在下で本宿主細菌は、得られた分解産物から有用化合物を発酵生産できる。これまで工業的に利用可能なBacillus属細菌が育種されているが、イオン液体はこれらの微生物の生育を阻害するため、実用的ではない。世界中でイオン液体耐性細菌の探索が進む中、独自のイオン液体耐性Bacillus属細菌とイオン液体を活用した有用物質変換系を開発することは、バイオマスの利用技術の開発につながる。