Development of E.coli cell for alkane production using a visualizing method of in vivo production of fatty aldehyde

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K20986
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry; Chemical biology
• Research Institution: The University of Tokyo
• Principal Investigator: Hayashi Yuuki 東京大学, 大学院総合文化研究科, 助教 (90444059)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: タンパク質 / 進化分子工学 / バイオエネルギー / アルカン合成酵素

Outline of Final Research Achievements

To practical alkane production in E.coli cells, we need to explore alkane synthesis enzymes, AAR and ADO, with higher activity, engineer a pathway to enhance a substrate (acyl-ACP) production and delete genes competing alkane production. At first, we compared catalytic activity of AAR and ADO derived from various cyanobacteria and successfully identified the highest active AAR and ADO. In parallel, we genetically engineered E.coli cells to emit bioluminescence depending on yield of aldehyde production in the cells. Using this engineered cells and a high-sensitive chemi-luminescence imager, we can evaluate amounts of aldehyde production in cells in real-time by monitoring bio-luminescent intensities emitted from over 1000 colonies on agar plate. This method enables high through-put screening to find gene disrupted strains, which enhanced acyl-ACP production, and/or decreased aldehyde consumption in competitive enzyme reaction, among library of strains deleted genes randomly.

Free Research Field

進化分子工学