Metabolic Engineering of Escherichia coli for High-Level Production of Lacto-N-neotetraose and Lacto-N-tetraose

• DOI: 10.1021/acs.jafc.3c02997
• Publication Date: 2023-07-19T21:36:03Z
• Authors: Yingxue Liao; Jinyong Wu; Zhongkui Li; Jin Wang; Lixia Yuan; Caiwen Lao; Xiangsong Chen; Jianming Yao
• Keywords: Metabolic Engineering; Galactosyltransferase; Strain (injury)

Lacto-N-neotetraose (LNnT) and lacto-N-tetraose (LNT) are important oligosaccharides found in breast milk commonly used as nutritional supplements infant formula. We metabolic engineering techniques to optimize the modified Escherichia coli BL21 star (DE3) strain for efficient synthesis of LNnT LNT using β-1,4-galactosyltransferase (HpgalT) from Helicobacter pylori β-1,3-galactosyltransferase (SewbdO) Salmonella enterica subsp. salamae serovar, respectively. Further, we optimized expression three key genes, lgtA, galE, HpgalT (SewbdO), synthesize or deleted several genes (ugd, ushA, agp, wcaJ, otsA, wcaC) block competition UDP-galactose pathway. The produced with a titer 22.07 48.41 g/L, respectively, supplemented batch culture, producing 0.41 0.73 g/L/h, strategies this study contribute development cell factories high-level their derivatives.