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

ZHU Yanbing; LI Hebin; NI Hui; LIU Jingwen; XIAO Anfeng; CAI Huinong

doi:10.1007/s13131-014-0534-2

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Article Navigation > Acta Oceanologica Sinica > 2014 > 33(12): 163-169

ZHU Yanbing, LI Hebin, NI Hui, LIU Jingwen, XIAO Anfeng, CAI Huinong. Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3[J]. Acta Oceanologica Sinica, 2014, 33(12): 163-169. doi: 10.1007/s13131-014-0534-2

Citation:

ZHU Yanbing, LI Hebin, NI Hui, LIU Jingwen, XIAO Anfeng, CAI Huinong. Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3[J]. Acta Oceanologica Sinica, 2014, 33(12): 163-169. doi: 10.1007/s13131-014-0534-2

Citation:

ZHU Yanbing, LI Hebin, NI Hui, LIU Jingwen, XIAO Anfeng, CAI Huinong. Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3[J]. Acta Oceanologica Sinica, 2014, 33(12): 163-169. doi: 10.1007/s13131-014-0534-2

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

doi: 10.1007/s13131-014-0534-2

1.
College of Food and Biological Engineering, Jimei University, Xiamen 361021, China;Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China;Research Center of Food Microbiology and Enzyme Engineering Technology of Fujian Province, Xiamen 361021, China
2.
Xiamen Medical College, Xiamen 361008, China
3.
College of Food and Biological Engineering, Jimei University, Xiamen 361021, China

Received Date: 2013-03-15
Rev Recd Date: 2013-09-13

Abstract

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℃ and 60℃. 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.
- Geobacillus sp.,
- superoxide dismutase,
- thermostability,
- purification

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References(62)

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