Clone, Expression and Characterization of the Thermostable ?-Galactosidase from Soil Metagenomic DNA of Taishan Mountain / 微生物学通报
Microbiology
; (12)2008.
Article
in Zh
| WPRIM
| ID: wpr-686114
Responsible library:
WPRO
ABSTRACT
A possible ?-Galactosidase gene(pwtsA) was discovered from soil metagenomic DNA of Taishan Mountain.PwtsA gene was inserted into the expression vector pET30a and transferred into E.coli BL21(DE3).Recombinant protein PWTSA was expressed as a soluble form at high level through IPTG induction,with a molecular mass of 57 kD analyzed by SDS-PAGE.PWTSA can produce o-nitrophenol from o-nitrophenol-?-D-galactopyranoside(ONPG),and its specific activity was determined as 13.6 U/mg.The enzymatic studies demonstrated that the recombinant protein PWTSA was a thermostable ?-Galactosidase,its optimum temperature and pH were 85?C~95?C and 6.5 respectively.In standard assays,the Km for ONPG was 0.83 mmol/L.
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WPRIM
Language:
Zh
Journal:
Microbiology
Year:
2008
Type:
Article