Functional expression of amylosucrase, a glucan-synthesizing enzyme, from Arthrobacter chlorophenolicus A6

J Microbiol Biotechnol. 2012 Sep;22(9):1253-7. doi: 10.4014/jmb.1201.01056.

Abstract

A gene (acas) designated as alpha-amylase was cloned from Arthrobacter chlorophenolicus A6. The multiple amino acid sequence analysis and functional expression of acas revealed that this gene really encoded an amylosucrase (ASase) instead of alpha-amylase. In fact, the recombinant enzyme exhibited typical ASase activity by showing both sucrose hydrolysis and glucosyltransferase activities. The purified enzyme has a molecular mass of 72 kDa and exhibits optimal hydrolysis activity at 45 degrees C and a pH of 8.0. The analysis of the oligomeric state of ACAS with gel permeation chromatography revealed that the ACAS existed as a monomer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arthrobacter / enzymology*
  • Arthrobacter / genetics
  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Glucosyltransferases / biosynthesis*
  • Glucosyltransferases / genetics
  • Glucosyltransferases / isolation & purification
  • Molecular Sequence Data
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Sequence Alignment
  • Sucrose / metabolism

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Sucrose
  • Glucosyltransferases
  • amylosucrase