Functional analyses of enzymes and the accessory proteins involved in industrial production of amide compounds

2020 Fiscal Year Final Research Report

• Project/Area Number: 17H03962
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied molecular and cellular biology
• Research Institution: University of Tsukuba
• Principal Investigator: HASHIMOTO Yoshiteru 筑波大学, 生命環境系, 准教授 (00323254)
• Co-Investigator(Kenkyū-buntansha): 小林 達彦 筑波大学, 生命環境系, 教授 (70221976) ; 白木 賢太郎 筑波大学, 数理物質系, 教授 (90334797)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 酵素 / 微生物

Outline of Final Research Achievements

Pseudomonas chlororaphis B23 yields nitrile hydratase (NHase) used for the production of 5- cyanovaleramide at the industrial level.
Here, we constructed several plasmids for the expression of NHases (and these mutant enzymes) in the presence and absence of each accessory protein of the NHase. Besides the expression of each of these NHases in E. coli carrying the resultant plasmid, we purified and characterized them. Based on the results of a series of experiments, we clarified the mechanisms for post-translational activation of NHase.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

今回の研究で対象としたB23株のニトリル水和酵素は、かつて(アクリロニトリルからの)アクリルアミド工業生産の第二世代触媒として使用され、また、シアノバレルアミド工業生産にも利用されてきた実用酵素である。本研究でのニトリル水和酵素のアクセサリータンパク質の解析により、実用酵素の活性化機構が明らかとなり、基礎的研究の見地からのみならず、（産業上有用な技術開発に貢献できる）応用的見地からも極めて意義深い。