Metabolic channeling with enzyme stapler

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K18276
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Kobe University
• Principal Investigator: Matsumoto Takuya 神戸大学, 科学技術イノベーション研究科, 特命助教 (40727161)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 大腸菌 / 代謝工学 / 酢酸 / 代謝チャネリング / 酵素

Outline of Final Research Achievements

We demonstrate metabolic enzyme ligation using a sortase A in the microbial cytoplasm for the redirection of metabolic flux through metabolic channeling. Here, sortase A expression was controlled by the lac promoter to trigger metabolic channeling by the addition of isopropyl-β-d-thiogalactopyranoside (IPTG). We tested covalent linking of pyruvate-formate lyase and phosphate acetyltransferase by sortase A-mediated ligation and evaluated the production of acetate. The time point of addition of IPTG was not critical for facilitating metabolic enzyme ligation, and acetate production increased upon expression of sortase A. These results show that sortase A-mediated enzyme ligation enhances an acetate-producing flux in E. coli. We have validated that sortase A-mediated enzyme ligation offers a metabolic channeling approach to redirect a central flux to a desired flux.

Free Research Field

生物化学工学