| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| 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.
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