Purification and biochemical characterization of m

Authors: Yanbing Zhu Hebin Li Hui Ni Jingwen Liu Anfeng Xiao Huinong Cai

Publish Date: 2014/10/13

Volume: 33, Issue: 12, Pages: 163-169

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Abstract

Thermostable SOD is a promising enzyme in biotechnological applications In the present study thermophile Geobacillus sp EPT3 was isolated from a deepsea hydrothermal field in the East Pacific A thermostable superoxide dismutase SOD from this strain was purified to homogeneity by steps of fractional ammonium sulfate precipitation DEAESepharose chromatography and PhenylSepharose chromatography SOD was purified 134 fold to homogeneity with a specific activity of 3 354 U/mg and 111 recovery SOD from Geobacillus sp EPT3 was of the MnSOD type judged by the insensitivity of the enzyme to both KCN and H2O2 SOD was determined to be a homodimer with monomeric molecular mass of 260 kDa It had high thermostability at 50°C and 60°C At tested conditions SOD was relatively stable in the presence of some inhibitors and denaturants such as βmercaptoethanol βME dithiothreitol DTT phenylmethylsulfonyl fluoride PMSF urea and guanidine hydrochloride Geobacillus sp EPT3 SOD showed striking stability across a wide pH range from 50 to 110 It could withstand denaturants of extremely acidic and alkaline conditions which makes it useful in the industrial applicationsFoundation item The National Natural Science Foundation of China under contract No 31371751 the Science and Technology Program of Xiamen China under contract No 201303120001 the Foundation for Innovative Research Team of Jimei University China under contract No 2010A006

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