Creation of new light-dependent enzymes using cyanobacteria with high transformation efficiency

2017 Fiscal Year Final Research Report

• Project/Area Number: 26660084
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: Nagoya University
• Principal Investigator: FUJITA Yuichi 名古屋大学, 生命農学研究科, 教授 (80222264)
• Research Collaborator: TSUJIMOTO Ryoma ; YAMAKAWA Hisanori ; HIRAIDE Yuto ; HASHIMOTO Kazuki
• Project Period (FY): 2014-04-01 – 2018-03-31
• Keywords: 光依存型プロトクロロフィリド還元酵素 / シアノバクテリア / クロロフィル生合成 / 酵素 / 進化

Outline of Final Research Achievements

Light-dependent protochlorophyllide reductase (LPOR), which requires light for catalysis, operates in chlorophyll biosynthesis. LPOR belongs to the short chain dehydrogenase (SDR) family and has an intrinsic loop structure that provides the structural basis for the light-driven catalytic function in addition to the common structure of the SDR family. In this study, we tried to re-create a new LPOR from a library constructed using synthetic semi-random DNA in the cyanobacterial mutant, in which shows a lethal phenotype under high light conditions in Synechocystis sp. PCC 6803. However, at the moment, a mutant carrying such a new LPOR has not yet isolated. We consider two main technical hurdles that should be solved in future; 1) semi-random DNA fragments have length variation (±2 bases) in current synthetic technology. 2) the parental strain contains a cell population bearing high-light tolerant revertants, which may be generated by high mutation rate in the recJ mutant.

Free Research Field

植物生化学