A Novel β-1,4-Galactosyltransferase from <i>Histophilus somni</i> Enables Efficient Biosynthesis of Lacto-<i>N</i>-Neotetraose via Both Enzymatic and Cell Factory Approaches
Guocong Luo, Yingying Zhu, Jiawei Meng, Li Wan, Wenli Zhang, Wanmeng Mu
Abstract
Human milk oligosaccharides (HMOs) attract particular attention because of their health benefits for infants. Lacto-N-neotetraose (LNnT) is one of the most abundant neutral core structures of HMOs. Bacterial β-1,4-galactosyltransferase (β-1,4-GalT) displays an irreplaceable role in the practical application of LNnT biosynthesis. In this study, a novel β-1,4-GalT from Histophilus somni was identified to efficiently synthesize LNnT from UDP-Gal and lacto-N-triose II (LNT II). The optimum pH and temperature were determined to be pH 6.0 and 30 °C, respectively. The enzyme showed both transgalactosylation and hydrolysis activity, with a specific activity of 3.7 and 6.6 U/mg, respectively. LNnT was synthesized using H. somni β-1,4-GalT via both enzymatic and cell factory approaches, and both approaches provided an LNnT ratio with the remaining LNT II at approximately 1:2 when reactions attained a balance. These findings indicated that H. somni β-1,4-GalT has a potential in biosynthesis of LNnT and derivatives in future.