[Expression, characterization and application of thermostable beta-glucuronidase from Thermotoga maritima]

Sheng Wu Gong Cheng Xue Bao. 2008 Aug;24(8):1407-12.
[Article in Chinese]

Abstract

The gene of beta-glucuronidase from Thermotoga maritima was cloned into the plasmid pHsh, and expressed in Escherichia coli JM109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 65.9 kD. The optimal activity of beta-glucuronidase was found at pH 5.0 and 80 degrees C. The purified enzyme was stable over a pH range from 5.8 to 8.2 and had a half life of 2 h at 80 degrees C. The kinetic experiments at 80 degrees C with p-nitrophenyl-beta- glucuronide as substrate gave a K(m) and V(max) of 0.18 mmol/L and 312 u per mg of protein. The purified enzyme could transform glycyrrhizin to glycyrrhetinic acid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Enzyme Stability
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Glucuronidase / biosynthesis*
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Glycyrrhetinic Acid / metabolism
  • Glycyrrhizic Acid / metabolism
  • Hot Temperature
  • Kinetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Thermotoga maritima / enzymology*
  • Thermotoga maritima / genetics*

Substances

  • Recombinant Proteins
  • Glycyrrhizic Acid
  • Glucuronidase
  • Glycyrrhetinic Acid