An iterative glycosyltransferase EntS catalyzes transfer and extension of O- and S-linked monosaccharide in enterocin 96

Abstract

Glycosyltransferases are essential tools for in vitro-glycoengineering. Bacteria harbor an unexplored variety of protein glycosyltransferases. Here, we describe a peptide glycosyltransferase (EntS) encoded by ORF0417 of Enterococcus faecalis TX0104. EntS di-glycosylates linear peptide of enterocin 96- a known antibacterial, in vitro. It is capable of transferring as well as extending the glycan onto the peptide in an iterative sequential dissociative manner. It can catalyze multiple linkages: Glc/Gal(-O)Ser/Thr, Glc/Gal(-S)Cys and Glc/Gal(β)Glc/Gal(-O/S)Ser/Thr/Cys, in one pot. Using EntS generated glycovariants of enterocin 96 peptide, size and identity of the glycan are found to influence bioactivity of the peptide. The study identifies EntS as an enzyme worth pursuing, for in vitro peptide glycoengineering.

Bacteriocin ASM1 is an O/S‐diglycosylated, plasmid‐encoded homologue of glycocin F

“…The two Enterococcus glycocins are modified by O‐linked sugars only , the two Bacillus and the Aeribacillus glycocins by an S‐linked glucose , and glycocin F is both O‐ and S‐GlcNAcylated . Protein sidechain glycosylation is generally considered to fulfill physicochemical and/or molecular recognition roles .…”

Using Chemical Synthesis to Probe Structure–Activity Relationships of the Glycoactive Bacteriocin Glycocin F

“…15 The only other bacteriocins that are unequivocally glycoactive are enterocin F4-9, which requires glycosylation on serines 46 and 38 for activity 20 and enterocin 96. 23 The NMR structure of GccF was determined in 2011, 25 followed by that of sublancin 168 in 2014, 26 confirming that these two glycocins share a similar core peptide scaffold, despite their different modes of action.…”

Protein Glycosylation in Bacterial Cells and Its Potential Applications

“…Jak dotąd S-glikozylowane białka opisano w komórkach m.in. Lactobacillus plantarum, Bacillus subtilis czy Enterococcus faecalis [79,85,108]. W glikoproteinach zidentyfikowanych w obrębie Bacteria i Archaea dominuje typ wiązania O-glikozydowego.…”