Synthetic redesign of central carbon and redox metabolism for high yield production of N-acetylglucosamine in Bacillus subtilis

枯草芽孢杆菌 产量(工程) 碳纤维 氧化还原 生物化学 化学 N-乙酰氨基葡萄糖 新陈代谢 生物 细菌 计算机科学 有机化学 材料科学 复合数 遗传学 冶金 算法
作者
Yang Gu,Xueqin Lv,Yanfeng Liu,Jianghua Li,Guocheng Du,Jian Chen,Rodrigo Ledesma‐Amaro,Long Liu
出处
期刊:Metabolic Engineering [Elsevier BV]
卷期号:51: 59-69 被引量:68
标识
DOI:10.1016/j.ymben.2018.10.002
摘要

Abstract One of the primary goals of microbial metabolic engineering is to achieve high titer, yield and productivity (TYP) of engineered strains. This TYP index requires optimized carbon flux toward desired molecule with minimal by-product formation. De novo redesign of central carbon and redox metabolism holds great promise to alleviate pathway bottleneck and improve carbon and energy utilization efficiency. The engineered strain, with the overexpression or deletion of multiple genes, typically can’t meet the TYP index, due to overflow of central carbon and redox metabolism that compromise the final yield, despite a high titer or productivity might be achieved. To solve this challenge, we reprogramed the central carbon and redox metabolism of Bacillus subtilis and achieved high TYP production of N-acetylglucosamine. Specifically, a “push–pull–promote” approach efficiently reduced the overflown acetyl-CoA flux and eliminated byproduct formation. Four synthetic NAD(P)-independent metabolic routes were introduced to rewire the redox metabolism to minimize energy loss. Implementation of these genetic strategies led us to obtain a B. subtilis strain with superior TYP index. GlcNAc titer in shake flask was increased from 6.6 g L−1 to 24.5 g L−1, the yield was improved from 0.115 to 0.468 g GlcNAc g−1 glucose, and the productivity was increased from 0.274 to 0.437 g L−1 h−1. These titer and yield are the highest levels ever reported and, the yield reached 98% of the theoretical pathway yield (0.478 g g−1 glucose). The synthetic redesign of carbon metabolism and redox metabolism represent a novel and general metabolic engineering strategy to improve the performance of microbial cell factories.
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