Expression and characterization of a novel trehalase from Microvirga sp. strain MC18

Protein Expr Purif. 2021 Jun:182:105846. doi: 10.1016/j.pep.2021.105846. Epub 2021 Feb 13.

Abstract

Trehalase catalyzes the hydrolysis of trehalose into two glucose molecules and is present in nearly all tissues in various forms. In this study, a putative bacterial trehalase gene, encoding a glycoside hydrolase family 15 (GH15) protein was identified in Microvirga sp. strain MC18 and heterologously expressed in E. coli. The specific activity of the purified recombinant trehalase MtreH was 24 U/mg, with Km and Vmax values of 23.45 mg/mL and 184.23 μmol/mg/min, respectively. The enzyme exhibited optimal activity at 40 °C and pH 7.0, whereby Ca2+ had a considerable positive effects on the catalytic activity and thermostability. The optimized enzymatic reaction conditions for the bioconversion of trehalose using rMtreH were determined as 40 °C, pH 7.0, 10 h and 1% trehalose concentration. The characterization of this bacterial trehalase improves our understanding of the metabolism and biological role of trehalose in prokaryotic organism.

Keywords: Enzymological characterization; Heterologous expression; Microvirga sp. strain MC18; Trehalase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins* / biosynthesis
  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / isolation & purification
  • Enzyme Stability
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression*
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Methylobacteriaceae* / enzymology
  • Methylobacteriaceae* / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Trehalase* / biosynthesis
  • Trehalase* / chemistry
  • Trehalase* / genetics
  • Trehalase* / isolation & purification

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Trehalase