2013 Fiscal Year Final Research Report
Construction of in vitro artificial metabolic pathway using thermo-tolerant enzymes
| Project/Area Number |
23780078
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied microbiology
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| Research Institution | Osaka University |
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Principal Investigator |
HONDA Kohsuke 大阪大学, 工学(系)研究科(研究院), 准教授 (90403162)
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| Project Period (FY) |
2011 – 2013
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| Keywords | 酵素反応 / 発酵 / 生体機能利用 / 代謝工学 |
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| Research Abstract |
An artificial pyruvate decarboxylation pathway, through which pyruvate can be converted to acetyl-CoA, was constructed by using a coupled enzyme system consisting of thermo-tolerant pyruvate decarboxylase and CoA-acylating aldehyde dehydrogenase. To demonstrate the applicability of the artificial pathway for chemical production, a cofactor-balanced and CoA-recycling synthetic pathway for N-acetylglutamate production was designed by coupling the acetyl-CoA-forming pathway with the glutamate dehydrogenase and N-acetylglutamate synthetase. N-Acetylglutamate could be produced from an equimolar mixture of pyruvate and alpha-ketoglutarate with a molar yield of 55% through the synthetic pathway consisting of the four thermo-tolerant enzymes. The overall recycling number of CoA was calculated to be 27.
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