Development of an efficient bio-production system using thermostable enzyme modules

2015 Fiscal Year Final Research Report

• Project/Area Number: 25820402
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Osaka University
• Principal Investigator: Okano Kenji 大阪大学, 工学(系)研究科(研究院), 助教 (40623335)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: In vitro代謝工学 / 耐熱性酵素 / 非リン酸化型Entner-Doudoroff経路 / 乳酸 / 人工オペロン

Outline of Final Research Achievements

Novel and efficient bio-production system using thermostable enzyme modules was developed. In this system, recombinant Escherichia coli strains having a heterologous thermophilic enzyme were directly used as catalysts at high temperatures. The denaturation of indigenous enzymes at high temperatures resulted in a one-step preparation of highly selective thermotolerant biocatalytic modules. The combination of these modules enabled to construct in vitro synthetic pathways specialized for chemical manufacture.
By using this technique, in this study, the non-phosphorylated Entner-Doudoroff glycolysis pathway, which was found in some hyperthermophile, was constructed. By coupling this pathway with lactate dehydrogenase, lactate was produced from glucose with 100% molar yield. Moreover, an artificial operon including the genes encoding eight thermophilic enzymes was successfully constructed and co-expression of eight enzymes in a single recombinant E. coli strains was achieved.

Free Research Field

生物化学工学