期刊
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
卷 85, 期 7, 页码 1772-1781出版社
OXFORD UNIV PRESS
DOI: 10.1093/bbb/zbab074
关键词
Escherichia coli; 3-fucosyllactose; cofactors; sorbitol; fructose-6-phosphate
类别
资金
- Science and Technology Service Network Initiative of Chinese Academy of Science [KFJ-STS-QYZD-122]
- Science and Technology Planning Project of Wuhan City [2020020602012119]
The metabolically engineered Escherichia coli effectively produced 3-fucosyllactose through the modified alpha-1,3-fucosyltransferase and the enhanced regeneration of cofactors (NADPH and GTP).
3-fucosyllactose (3-FL) and 2'-fucosyllactose (2'-FL), are two important fucosylated oligosaccharides in human milk. Extensive studies on 2'-FL enabled its official approval for use in infant formula. However, development of 3-FL has been somewhat sluggish due to its low content in human milk and poor yield in enlarged production. Here, an alpha-1,3-fucosyltransferase mutant was introduced into an engineered Escherichia coli (E. coli) capable of producing GDP-L-fucose, leading to a promising 3-FL titer in a 5.0-L bioreactor. To increase the availability of cofactors (NADPH and GTP) for optimized 3-FL production, zwf, pntAB, and gsk genes were successively overexpressed, finally resulting in a higher 3-FL level with a titer of 35.72 g/L and a yield of 0.82 mol 3-FL/mol lactose. Unexpectedly, the deletion of pfkA gene led to a much lower performance of 3-FL production than the control strain. Still, our strategy achieved the highest 3-FL level in E. coli to date. [GRAPHICS] The metabolically engineered Escherichia coli effectively produced 3-fucosyllactose through the modified alpha-1,3-fucosyltransferase and the enhanced regeneration of cofactors (NADPH and GTP).
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