Cloning, purification and characterization of a thermostable amylosucrase from Deinococcus geothermalis

FEMS Microbiol Lett. 2008 Aug;285(1):25-32. doi: 10.1111/j.1574-6968.2008.01204.x. Epub 2008 Jun 3.

Abstract

Amylosucrase is a transglucosidase that catalyses the synthesis of an amylose-type polymer from sucrose, an abundant agro-resource. Here we describe a novel thermostable amylosucrase from the moderate thermophile Deinococcus geothermalis (DGAS). The dgas gene was cloned and expressed in Escherichia coli. The encoded enzyme was purified and characterized. DGAS displays a specific activity of 44 U mg(-1), an optimal temperature of 50 degrees C and a half-life of 26 h at 50 degrees C. Moreover, it produces an alpha-glucan at 50 degrees C, with an average degree of polymerization of 45 and a polymerization yield of 76%. DGAS is thus the most active and thermostable amylosucrase known to date.

MeSH terms

  • Amino Acid Sequence
  • Amylose / metabolism
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Bacterial Proteins / metabolism
  • Cloning, Molecular*
  • Deinococcus / chemistry
  • Deinococcus / enzymology*
  • Deinococcus / genetics
  • Enzyme Stability
  • Gene Expression
  • Glucosyltransferases / chemistry*
  • Glucosyltransferases / genetics
  • Glucosyltransferases / isolation & purification*
  • Glucosyltransferases / metabolism
  • Molecular Sequence Data
  • Sequence Alignment
  • Temperature

Substances

  • Bacterial Proteins
  • Amylose
  • Glucosyltransferases
  • amylosucrase