Development of a 2-pyrrolidone biosynthetic pathway in Corynebacterium glutamicum by engineering an acetyl-CoA balance route

被引:3
|
作者
Xu, Meijuan [1 ]
Gao, Hui [1 ]
Ma, Zhenfeng [1 ]
Han, Jin [1 ]
Zheng, Keyi [2 ]
Shao, Minglong [1 ]
Rao, Zhiming [1 ]
机构
[1] Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, 1800 Lihu Rd, Wuxi 214122, Jiangsu, Peoples R China
[2] Meihua Biotechnol Grp Co, Wujiaqu 831300, Peoples R China
关键词
Corynebacterium glutamicum; 2-Pyrrolidone; 5 ' Untranslated region (UTR); Acetyl-CoA regeneration; Metabolic engineering; ESCHERICHIA-COLI; GLUTAMATE-DECARBOXYLASE; EXPRESSION; GAD;
D O I
10.1007/s00726-022-03174-0
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
2-Pyrrolidone is widely used in the textile and pharmaceutical industries. Here, we established a 2-pyrrolidone biosynthesis pathway in Corynebacterium glutamicum, by expressing glutamate decarboxylase (Gad) mutant and beta-alanine CoA transferase (Act) which activates spontaneous dehydration cyclization of GABA to form 2-pyrrolidone. Also, the 5 ' untranslated regions (UTR) strategy was used to increase the expression of protein. Furthermore, considering the importance of acetyl-CoA in the 2-pyrrolidone synthesis pathway, the acetyl-CoA synthetase (acsA) gene was introduced to convert acetate into acetyl-CoA thus achieving the recyclability of the economy. Finally, the fed-batch fermentation of the final strain in a 5 L bioreactor produced 10.5 g/L 2-pyrrolidone within 78 h, which increased by 42.5% by altering the level of gene expression. This is the first time to build the basic chemical 2-pyrrolidone from glucose in one step in C. glutamicum.
引用
收藏
页码:1437 / 1450
页数:14
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