Screening of novel carbohydrate phosphorylase using metagenomics

Research Project

Project/Area Number	16K07688
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Applied biochemistry
Research Institution	Niigata University
Principal Investigator	NAKAI Hiroyuki 新潟大学, 自然科学系, 准教授 (00400002)
Project Period (FY)	2016-04-01 – 2019-03-31
Project Status	Completed (Fiscal Year 2018)
Budget Amount *help	¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000) Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords	ホスホリラーゼ / オリゴ糖 / 酵素 / 糖
Outline of Final Research Achievements	Phosphorylases are exolytic enzymes catalyzing phosphorolysis of particular glycosides to produce sugar 1-phosphate with strict substrate specificity. The reaction is reversible, enabling the practical synthesis of oligosaccharides. However, there is little variation among phosphorylases and this limits their utilization for the production of oligosaccharides. Therefore, it would be beneficial to identify phosphorylases with previously unreported substrate specificities. In this study, we developed a metagenomics screening method specialized for novel phosphorylase using E. coli with altered carbohydrate utilization.
Academic Significance and Societal Importance of the Research Achievements	生体内糖質代謝に関与する糖質加リン酸分解酵素（ホスホリラーゼ）によるオリゴ糖合成の収率は高く、産業上有用な酵素になり得る。今回新規ホスホリラーゼの探索に特化したタゲノミクススクリーニング手法を開発できたことで、今後これまで報告例のない新たな基質特異性を有する酵素の発見および合成可能なオリゴ糖のバリエーション拡大が期待される。また、新規ホスホリラーゼの発見は当該酵素が関与する新たな生体内糖質代謝機構の解明にも繋がり、生物学的意義からも学術的に重要な研究となる。