Metabolic Engineering of Escherichia coil for Efficient Biosynthesis of Lacto-N-tetraose Using a Novel β-1,3-Galactosyltransferase from Pseudogulbenkiania ferrooxidans

被引:30
作者
Zhu, Yingying [1 ]
Li, Zeyu [1 ]
Luo, Guocong [1 ]
Wu, Hao [1 ]
Zhang, Wenli [1 ]
Mu, Wanmeng [1 ,2 ]
机构
[1] Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China
[2] Jiangnan Univ, Int Joint Lab Food Safety, Wuxi 214122, Jiangsu, Peoples R China
基金
中国国家自然科学基金;
关键词
biosynthesis; beta-1,3-galactosyltrans erase; human milk oligosaccharides; lacto-N-tetraose; lacto-N-triose II; HUMAN-MILK OLIGOSACCHARIDES; FORMULA; GROWTH; ENZYME; COLI;
D O I
10.1021/acs.jafc.1c04059
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
Human milk oligosaccharides (HMOs) attract considerable interest in recent years because of their particular role in infant health. Lacto-N-tetraose (LNT), one of the most abundant HMOs, has been commercially added in the infant formula as a functional fortifier. In this study, a novel LNT-producing beta-1,3-galactosyltransferase (beta-1,3-GalT) from Pseudogulbenkiania ferrooxidans was screened from 14 putative candidates, and a highly LNT-producing metabolically engineered Escherichia coli strain was constructed based on a previously constructed lacto-N-triose II (LNT II)-producing strain, by strengthening UDP-galactose synthesis and introduction of P. ferrooxidans beta-1,3-GalT. The engineered strain produced 3.11 and 25.49 g/L LNT in shake-flask and fed-batch cultivation, with the molar conversion ratio of LNT II to LNT of 88.15 and 85.09%, respectively. The productivity and specific yield of LNT in fed-batch cultivation were measured to be 0.61 g/L.h and 0.76 g/g dry cell weight, respectively. To the best of our knowledge, it is the highest LNT yield ever reported.
引用
收藏
页码:11342 / 11349
页数:8
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