Expression, characterization and application of thermostable beta-glucuronidase from Thermotoga maritima.
- Author:
Zhuo WANG
1
;
Jianjun PEI
;
Huazhong LI
;
Weilan SHAO
Author Information
1. School of Biotechnology, Jiangnan University, Wuxi 214122, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Enzyme Stability;
Escherichia coli;
enzymology;
genetics;
Glucuronidase;
biosynthesis;
genetics;
metabolism;
Glycyrrhetinic Acid;
metabolism;
Glycyrrhizic Acid;
metabolism;
Hot Temperature;
Kinetics;
Recombinant Proteins;
biosynthesis;
genetics;
metabolism;
Thermotoga maritima;
enzymology;
genetics
- From:
Chinese Journal of Biotechnology
2008;24(8):1407-1412
- CountryChina
- Language: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.
