[Purification and characterization of a sarcosine oxidase from Bacillus sp. BSD-8].
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.
College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
Hui
H
Liu
Guiqin
G
Sun
Xiaohang
X
Ma
Lingyan
L
Sun
Xiangfeng
X
Lu
Pengcheng
P
Zhang
D001407:Bacillus / Q000201:enzymology* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011232:Chemical Precipitation; D004795:Enzyme Stability; D050879:Sarcosine Oxidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012988:Soil Microbiology
D001426:Bacterial Proteins; D050879:Sarcosine Oxidase