Cloning, expression and functional analysis of the genes in TPS/TPP trehalose synthetic pathway of Meiothermus ruber.
- Author:
Yueming ZHU
1
;
Yichen TANG
;
Hengyi XU
;
Juan ZHANG
;
Dongsheng WEI
;
Laijun XING
;
Mingchun LI
Author Information
1. Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin 300071, China.
- Publication Type:Journal Article
- MeSH:
Bacterial Proteins;
genetics;
metabolism;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Glucosyltransferases;
genetics;
metabolism;
Phosphoric Monoester Hydrolases;
genetics;
metabolism;
Recombinant Fusion Proteins;
genetics;
isolation & purification;
metabolism;
Thermus;
enzymology;
genetics;
Trehalose;
biosynthesis
- From:
Chinese Journal of Biotechnology
2009;25(3):399-405
- CountryChina
- Language:Chinese
-
Abstract:
By constructing the genomic DNA library of Meiothermus ruber CBS-01, the genes of trehalose phosphate synthase (TPS) and trehalose phosphate phosphatase (TPP) involved in trehalose synthesis were cloned. The genes were cloned into the plasmid pET21a, and expressed in Escherichia coli Rosetta gami (DE3). The activities of these two purified enzymes were confirmed by thin layer chromatography (TLC). Meanwhile, we tested the cellular compatible solutes of M. ruber CBS-01 under different environmental pressure, and found that under hyperosmotic pressure, this strain can accumulate trhalose-6-phosphate, but not trehalose. These results can give more insight to future research in the roles of TPS/TPP and TreS pathway.
