Purification and characterization of a sarcosine oxidase from Bacillus sp. BSD-8.
- Author:
Hui LIU
1
;
Guiqin SUN
;
Xiaohang MA
;
Lingyan SUN
;
Xiangfeng LU
;
Pengcheng ZHANG
Author Information
1. College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
- Publication Type:Journal Article
- MeSH:
Bacillus;
enzymology;
isolation & purification;
Bacterial Proteins;
chemistry;
isolation & purification;
metabolism;
Chemical Precipitation;
Enzyme Stability;
Sarcosine Oxidase;
chemistry;
isolation & purification;
metabolism;
Soil Microbiology
- From:
Chinese Journal of Biotechnology
2010;26(3):335-340
- CountryChina
- Language:Chinese
-
Abstract:
We purified a sarcosine oxidase from Bacillus sp. strain BSD-8 isolated from soil. We purified the enzyme by ammonium sulfate precipitation, DEAE-cellulose, Toyopearl hydrophobic and Sephadex G-75 molecular sieve chromatography and characterized the purified sarcosine oxidase. This sarcosine oxidase was a flavin enzyme containing a noncovalently bound flavin with the subunit molecular mass of 51 kDa. The optimal temperature for this enzyme was 60 degrees C and it showed its highest activity at pH 8.5. It was stable in the pH range of 8.0-10.0 and at the temperature of 60 degrees C. Estimated by Lineveaver-Burk plots, the K(m) of the enzyme was 3.1 mmol/L. Ag+, Hg2+, SDS and Tween 80 dramatically inhibted the enzyme activity, whereas Tween 20 and Triton X-100 had no effect on enzyme activity. The thermostability of this enzyme was better than reported sarcosine oxidases, and it could be applied in enzymatic measuring of creatinine.
