[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.
Liu
Hui
H
College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
Sun
Guiqin
G
Ma
Xiaohang
X
Sun
Lingyan
L
Lu
Xiangfeng
X
Zhang
Pengcheng
P
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