Wu Yan, Chu Wanying, Yang Jiayao, Xu Yudong, Shen Qi, Yang Haoning, Xu Fangxu, Liu Yefei, Lu Ping, Jiang Ke, Zhao Hongxin
Zhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Key Laboratory of Urban Agriculture by Ministry of Agriculture of China, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
Front Microbiol. 2021 Oct 8;12:754306. doi: 10.3389/fmicb.2021.754306. eCollection 2021.
Biotechnological production of 2,3-butanediol (2,3-BD), a versatile platform bio-chemical and a potential biofuel, is limited due to by-product toxicity. In this study, we aimed to redirect the metabolic flux toward 2,3-BD in () by increasing the intracellular NADH pool. Increasing the NADH/NAD ratio by knocking out the NADH dehydrogenase genes (/) enhanced 2,3-BD production by up to 67% compared with wild-type . When lactate dehydrogenase () was knocked out, the yield of 2,3-BD was increased by 71.2% compared to the wild type. Metabolic flux analysis revealed that upregulated expression of the sRNA RyhB led to a noteworthy shift in metabolism. The 2,3-BD titer of the best mutant Ea-2 was almost seven times higher than that of the parent strain in a 5-L fermenter. In this study, an effective metabolic engineering strategy for improved 2,3-BD production was implemented by increasing the NADH/NAD ratio and blocking competing pathways.
2,3-丁二醇（2,3-BD）是一种用途广泛的平台生物化学品和潜在生物燃料，其生物技术生产因副产物毒性而受到限制。在本研究中，我们旨在通过增加细胞内NADH库，将代谢通量重定向至（）中的2,3-丁二醇。与野生型（）相比，通过敲除NADH脱氢酶基因（/）提高NADH/NAD比率，使2,3-丁二醇产量提高了67%。当敲除乳酸脱氢酶（）时，与野生型相比，2,3-丁二醇产量提高了71.2%。代谢通量分析表明，sRNA RyhB的上调表达导致了代谢的显著转变。在5-L发酵罐中，最佳突变体Ea-2的2,3-丁二醇滴度几乎比亲本菌株高七倍。在本研究中，通过提高NADH/NAD比率和阻断竞争途径，实施了一种有效的代谢工程策略来提高2,3-丁二醇的产量。
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