Chemoenzymatic synthesis of fucosylated oligosaccharides using Thermosynechococcus α1-2-fucosyltransferase and their application in the regulation of intestinal microbiota.

ABSTRACT: A novel bacterial α 1-2-fucosyltransferase (α 2FT) from Thermosynechococcus sp. NK55a (Ts2FT) has been discovered and characterized. It shares 28-62% protein sequence homology to α 2FTs reported previously. The Ts2FT was cloned as an N-terminal His₆-tagged recombinant protein (His₆-Ts2FT) and expressed in E. coli BL21 (DE3). It was expressed at a level of 6.2 mg/L culture after induction with 0.05 mM of isopropylβ-d-1-thiogalactoside (IPTG) at 16 °C for 20 h. It showed the optimal activity at a reaction temperature of 40 °C and pH of 7.0. The presence of a Mg²⁺ improved its catalytic efficiency. Ts2FT displayed a strict acceptor specificity and could recognize only β1-3-galatoside acceptors. It was used efficiently

SUBMITTER: Zhong R 

PROVIDER: S-EPMC8593560 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature