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Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3

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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 deep-sea 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, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD from Geobacillus sp. EPT3 was of the Mn-SOD 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 26.0 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 β-mercaptoethanol (β-ME), dithiothreitol (DTT), phenylmethylsulfonyl fluoride (PMSF), urea, and guanidine hydrochloride. Geobacillus sp. EPT3 SOD showed striking stability across a wide pH range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications.

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Authors and Affiliations

  1. College of Food and Biological Engineering, Jimei University, Xiamen, 361021, China

    Yanbing Zhu, Hui Ni, Jingwen Liu, Anfeng Xiao & Huinong Cai

  2. Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen, 361021, China

    Yanbing Zhu, Hui Ni, Anfeng Xiao & Huinong Cai

  3. Research Center of Food Microbiology and Enzyme Engineering Technology of Fujian Province, Xiamen, 361021, China

    Yanbing Zhu, Hui Ni, Anfeng Xiao & Huinong Cai

  4. Xiamen Medical College, Xiamen, 361008, China

    Hebin Li

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  1. Yanbing Zhu
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  2. Hebin Li
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  3. Hui Ni
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Correspondence to Huinong Cai.

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Foundation 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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Zhu, Y., Li, H., Ni, H. et al. Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3. Acta Oceanol. Sin. 33, 163–169 (2014). https://doi.org/10.1007/s13131-014-0534-2

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