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首页> 外文期刊>Applied Microbiology and Biotechnology >Engineering Escherichia coli with acrylate pathway genes for propionic acid synthesis and its impact on mixed-acid fermentation
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Engineering Escherichia coli with acrylate pathway genes for propionic acid synthesis and its impact on mixed-acid fermentation

机译：利用丙烯酸途径基因工程化丙酸合成的大肠杆菌及其对混合酸发酵的影响

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摘要

Fermentation-derived products are in greater demand to meet the increasing global market as well as to overcome environmental problems. In this work, Escherichia coli has been metabolically engineered with acrylate pathway genes from Clostridium propionicum for the conversion of d-lactic acid to propionic acid. The introduced synthetic pathway consisted of seven genes encoding the enzymes propionate CoA-transferase (Pct), lactoyl-CoA dehydratase (Lcd) and acryloyl-CoA reductase (Acr). The engineered strain synthesised propionic acid at a concentration of 3.7 ± 0.2 mM upon fermentation on glucose. This low production level could be attributed to the low activity of the recombinant enzymes in particular the rate-limiting enzyme, Acr. Interestingly, the recombinant pathway caused an increased lactate production in E. coli with a yield of 1.9 mol/mol of glucose consumed along with a decrease in other by-products. Down-regulation of the pfl (pyruvate formate lyase) genes and a possible inhibition of Pfl activity by the acrylate pathway intermediate, acryloyl-CoA, could have reduced carbon flow to the Pfl pathway with a concomitant increase in lactate production. This study reports a novel way of synthesising propionic acid by employing a non-native, user-friendly organism through metabolic engineering.
机译：为了满足不断增长的全球市场以及克服环境问题，对源自发酵的产品有更大的需求。在这项工作中，大肠杆菌已经用丙酸梭菌的丙烯酸酯途径基因进行了代谢改造，以将D-乳酸转化为丙酸。引入的合成途径由七个编码丙酸CoA转移酶（Pct），乳酰辅酶A脱水酶（Lcd）和丙烯酰辅酶A还原酶（Acr）的基因组成。在葡萄糖上发酵后，该工程菌株合成了浓度为3.7±0.2 mM的丙酸。这种低产量水平可归因于重组酶特别是限速酶Acr的低活性。有趣的是，重组途径导致大肠杆菌中乳酸产量增加，消耗的葡萄糖产量为1.9 mol / mol，同时其他副产物减少。 pfl（丙酮酸甲酸酯裂解酶）基因的下调和由丙烯酸酯途径中间体丙烯酰辅酶A可能对Pfl活性的抑制可能会减少流向Pfl途径的碳，同时增加乳酸的产生。这项研究报告了通过代谢工程采用非天然，用户友好的生物体来合成丙酸的新方法。

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