| 论文题目: | Metabolic engineering and flux analysis of for Corynebacterium glutamicum for L-serine production |
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| 作者: | Lai, SJ; Zhang, Y; Liu, SW; Liang, Y; Shang, XL; Chai, X; Wen, TY |
| 联系作者: | Wen, TY |
| 刊物名称: | SCIENCE CHINA-LIFE SCIENCES |
| 期: | 4 |
| 卷: | 55 |
| 页: | 283-290 |
| 年份: | 2012 |
| 影响因子: | 2.024 |
| 论文下载: | 下载地址 |
| 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. |
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