Metabolic Engineering of Enterobacter aerogenes for Improved 2,3-Butanediol Production by Manipulating NADH Levels and Overexpressing the Small RNA RyhB.
Metabolic Engineering of Enterobacter aerogenes for Improved 2,3-Butanediol Production by Manipulating NADH Levels and Overexpressing the Small RNA RyhB.
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DOI：
10.3389/fmicb.2021.754306
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发表时间：
2021
影响因子：
5.2
通讯作者：
Zhao H
中科院分区：
文献类型：
--
作者：
Wu Y;Chu W;Yang J;Xu Y;Shen Q;Yang H;Xu F;Liu Y;Lu P;Jiang K;Zhao H
2,3-丁二醇（2,3- bd）是一种多用途的生物化学平台和潜在的生物燃料，由于副产品毒性而受到限制。在这项研究中，我们旨在通过增加细胞内NADH池，将产气肠杆菌（E. aerogenes）的代谢通量转向2,3- bd。通过敲除NADH脱氢酶基因（nuoC/nuoD）提高NADH/NAD+比值，与野生型产气大肠杆菌相比，2,3- bd产量可提高67%。敲除乳酸脱氢酶（ldh）后，2,3- bd的产量比野生型提高了71.2%。代谢通量分析显示，上调sRNA RyhB的表达导致了代谢的显著变化。在5-L发酵条件下，最佳突变株Ea-2的2,3- bd滴度比亲本菌株高出近7倍。在本研究中，通过提高NADH/NAD+比率和阻断竞争途径，实现了一种有效的代谢工程策略，以提高2,3- bd的产量。
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 Enterobacter aerogenes (E. aerogenes) by increasing the intracellular NADH pool. Increasing the NADH/NAD+ ratio by knocking out the NADH dehydrogenase genes (nuoC/nuoD) enhanced 2,3-BD production by up to 67% compared with wild-type E. aerogenes. When lactate dehydrogenase (ldh) 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.
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影响因子：
3.2
作者：
BLOMQVIST, K;NIKKOLA, M;PENTTILA, ME
通讯作者：
PENTTILA, ME
影响因子：
3.9
作者：
Lyu, Yu;Wu, Jihui;Shi, Yunyu
通讯作者：
Shi, Yunyu
DOI：
10.1007/s004490000210
发表时间：
2000-12-01
期刊：
BIOPROCESS ENGINEERING
影响因子：
--
作者：
Perego, P;Converti, A;Canepa, P
通讯作者：
Canepa, P
影响因子：
6.7
作者：
Acuna, Lillian G.;Jose Barros, M.;Calderon, Ivan L.
通讯作者：
Calderon, Ivan L.
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL