Metabolic engineering and flux analysis of for Corynebacterium glutamicum for L-serine production

被引:28
作者
Lai ShuJuan [1 ,2 ]
Zhang Yun [1 ]
Liu ShuWen [1 ,2 ]
Liang Yong [1 ]
Shang XiuLing [1 ,2 ]
Chai Xin [1 ,2 ]
Wen TingYi [1 ]
机构
[1] Chinese Acad Sci, Inst Microbiol, Dept Ind Microbiol & Biotechnol, Beijing 100101, Peoples R China
[2] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
基金
中国国家自然科学基金;
关键词
Corynebacterium glutamicum; L-serine; intracellular metabolites; metabolic engineering; elementary mode analysis; ESCHERICHIA-COLI; SACCHAROMYCES-CEREVISIAE; METHIONINE PRODUCTION; HYDROXYMETHYLTRANSFERASE; BIOSYNTHESIS; PATHWAY; IDENTIFICATION; ACCUMULATION; INHIBITION; SEQUENCE;
D O I
10.1007/s11427-012-4304-0
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
L-Serine plays a critical role as a building block for cell growth, and thus it is difficult to achieve the direct fermentation of L-serine from glucose. In this study, ATCC 13032 was engineered by blocking and attenuating the conversion of L-serine to pyruvate and glycine, releasing the feedback inhibition by L-serine to 3-phosphoglycerate dehydrogenase (PGDH), in combination with the co-expression of 3-phosphoglycerate kinase (PGK) and feedback-resistant PGDH (PGDH(r)). The resulting strain, SER-8, exhibited a lower specific growth rate and significant differences in L-serine levels from Phase I to Phase V as determined for fed-batch fermentation. The intracellular L-serine pool reached (14.22 +/- 1.41) mu mol g(CDM)(-1), which was higher than glycine pool, contrary to fermentation with the wild-type strain. Furthermore, metabolic flux analysis demonstrated that the over-expression of PGK directed the flux of the pentose phosphate pathway (PPP) towards the glycolysis pathway (EMP), and the expression of PGDH(r) improved the L-serine biosynthesis pathway. In addition, the flux from L-serine to glycine dropped by 24%, indicating that the deletion of the activator GlyR resulted in down-regulation of serine hydroxymethyltransferase (SHMT) expression. Taken together, our findings imply that L-serine pool management is fundamental for sustaining the viability of , and improvement of C-1 units generation by introducing the glycine cleavage system (GCV) to degrade the excessive glycine is a promising target for L-serine production in C. glutamicum.
引用
收藏
页码:283 / 290
页数:8
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