Enhanced 3-Hydroxypropionic Acid Production From Acetate via the Malonyl-CoA Pathway in Corynebacterium glutamicum.
Enhanced 3-Hydroxypropionic Acid Production From Acetate via the Malonyl-CoA Pathway in Corynebacterium glutamicum.
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通过谷氨酸棒杆菌中的丙二酰辅酶A途径增强乙酸盐的3-羟基丙酸生产
DOI：
10.3389/fbioe.2021.808258
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发表时间：
2021
影响因子：
5.7
通讯作者：
Chen T
中科院分区：
文献类型：
--
作者：
Chang Z;Dai W;Mao Y;Cui Z;Zhang Z;Wang Z;Ma H;Chen T
醋酸盐是一种经济环保的碳源替代品。本文探讨了利用谷氨酸棒状杆菌作为宿主从乙酸中提取3-羟基丙酸（3-HP）的潜力。首先，通过多种策略优化金银花丙二酰辅酶a还原酶的表达水平，菌株Cgz2/sod-N-C*在瓶中MCR酶活性为63 nmol/mg/min， 3 hp滴度为0.66 g/L。接下来，通过削弱Cgz2/sod-N-C*中柠檬酸合成酶的表达，减少TCA循环中乙酰辅酶a的消耗，得到的菌株Cgz12/sod-N-C*从9.32 g/L乙酸中产生2.39 g/L 3-HP。然而，随后在Cgz12/sod-N-C*中乙酰辅酶a羧化酶基因表达的解除导致细胞内脂肪酸积累增加，而不是3-HP。因此，利用蓝草苷抑制Cgz14/sod-N-C*中脂肪酸的合成，其3-HP滴度进一步提高到4.26 g/L，产率为0.50 g 3-HP/g-acetate。最后，工程菌株在不添加蓝绿蛋白的情况下，在生物反应器中积累了17.1 g/L 3-HP，这是以乙酸为底物获得的最高滴度。结果表明，谷氨酸棒状杆菌是醋酸生产3-HP的理想宿主。
Acetate is an economical and environmental-friendly alternative carbon source. Herein, the potential of harnessing Corynebacterium glutamicum as a host to produce 3-hydroxypropionic acid (3-HP) from acetate was explored. First, the expression level of malonyl-CoA reductase from Chloroflexus aurantiacus was optimized through several strategies, strain Cgz2/sod-N-C* showed an MCR enzyme activity of 63 nmol/mg/min and a 3-HP titer of 0.66 g/L in flasks. Next, the expression of citrate synthase in Cgz2/sod-N-C* was weakened to reduce the acetyl-CoA consumption in the TCA cycle, and the resulting strain Cgz12/sod-N-C* produced 2.39 g/L 3-HP from 9.32 g/L acetate. However, the subsequent deregulation of the expression of acetyl-CoA carboxylase genes in Cgz12/sod-N-C* resulted in an increased accumulation of intracellular fatty acids, instead of 3-HP. Accordingly, cerulenin was used to inhibit fatty acid synthesis in Cgz14/sod-N-C*, and its 3-HP titer was further increased to 4.26 g/L, with a yield of 0.50 g 3-HP/g-acetate. Finally, the engineered strain accumulated 17.1 g/L 3-HP in a bioreactor without cerulenin addition, representing the highest titer achieved using acetate as substrate. The results demonstrated that Corynebacterium glutamicum is a promising host for 3-HP production from acetate.
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