EpsN from Bacillus subtilis 168 has UDP-2,6-dideoxy 2-acetamido 4-keto glucose aminotransferase activity in vitro

Glycobiology. 2018 Oct 1;28(10):802-812. doi: 10.1093/glycob/cwy063.

Abstract

The gene epsN of Bacillus subtilis 168 was cloned and overexpressed in Escherichia coli. Purified recombinant EpsN is shown to be a pyridoxal 5'-phosphate (PLP)-dependent aminotransferase by absorption spectroscopy, l-cycloserine inhibition and reverse phase HPLC studies. EpsN catalyzes the conversion of UDP-2,6-dideoxy 2-acetamido 4-keto glucose to UDP-2,6-dideoxy 2-acetamido 4-amino glucose. Lys190 was found by sequence comparison and site-directed mutagenesis to form Schiff base with PLP. Mutagenesis studies showed that, in addition to Lys190, Ser185, Glu164, Gly58 and Thr59 are essential for aminotransferase activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology*
  • Bacillus subtilis / metabolism
  • Bacterial Proteins / metabolism*
  • Biocatalysis
  • Glucose / analogs & derivatives*
  • Glucose / chemistry
  • Glucose / metabolism
  • Molecular Structure
  • Mutagenesis, Site-Directed
  • Polysaccharides, Bacterial / genetics
  • Polysaccharides, Bacterial / isolation & purification
  • Polysaccharides, Bacterial / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Transaminases / metabolism*
  • Uridine Diphosphate / chemistry
  • Uridine Diphosphate / metabolism*

Substances

  • Bacterial Proteins
  • Polysaccharides, Bacterial
  • Recombinant Proteins
  • UDP-2,6-dideoxy 2-acetamido 4-keto glucose
  • Uridine Diphosphate
  • Transaminases
  • Glucose