Redirection of pyruvate flux toward desired metabolic pathways through substrate channeling between pyruvate kinase and pyruvate-converting enzymes in Saccharomyces cerevisiae

被引：16

作者：

Kim, Sujin ^{[1
]}

Bae, Sang-Jeong ^{[1
]}

Hahn, Ji-Sook ^{[1
]}

机构：

[1] Seoul Natl Univ, Inst Chem Proc, Sch Chem & Biol Engn, 1 Gwanak Ro, Seoul 08826, South Korea

来源：

SCIENTIFIC REPORTS | 2016年 / 6卷

基金：

新加坡国家研究基金会;

关键词：

D-LACTIC ACID; LACTATE-DEHYDROGENASE; EFFICIENT PRODUCTION; ESCHERICHIA-COLI; YEAST; 2,3-BUTANEDIOL; SCAFFOLD; BIOSYNTHESIS; ORGANIZATION; IMPROVES;

D O I：

10.1038/srep24145

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Spatial organization of metabolic enzymes allows substrate channeling, which accelerates processing of intermediates. Here, we investigated the effect of substrate channeling on the flux partitioning at a metabolic branch point, focusing on pyruvate metabolism in Saccharomyces cerevisiae. As a platform strain for the channeling of pyruvate flux, PYK1-Coh-Myc strain was constructed in which PYK1 gene encoding pyruvate kinase is tagged with cohesin domain. By using high-affinity cohesin-dockerin interaction, the pyruvate-forming enzyme Pyk1 was tethered to heterologous pyruvate-converting enzymes, lactate dehydrogenase and a-acetolactate synthase, to produce lactic acid and 2,3-butanediol, respectively. Pyruvate flux was successfully redirected toward desired pathways, with a concomitant decrease in ethanol production even without genetic attenuation of the ethanol-producing pathway. This pyruvate channeling strategy led to an improvement of 2,3-butanediol production by 38%, while showing a limitation in improving lactic acid production due to a reduced activity of lactate dehydrogenase by dockerin tagging.

引用

页数：9

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