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Characterization of a thermostable manganese-containing superoxide dismutase from inshore hot spring thermophile Thermus sp. JM1

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Abstract

A thermostable superoxide dismutase (SOD) from the inshore thermophile Thermus sp. JM1 was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography and Phenyl-Sepharose chromatography. The specific activity of the purified native enzyme was 1 656 U/mg. A sod gene from this strain was cloned and overexpressed in Escherichia coli (E. coli). The prepared apo-enzyme of the purified recombinant SOD (rSOD) was reconstituted with either Fe or Mn by means of incubation with appropriate metal salts. As a result, only Mn2+-reconstituted rSOD (Mn-rSOD) exhibited the specific activity of 1 598 U/mg. SOD from Thermus sp. JM1 was Mn-SOD, judging by the specific activities analysis of Fe or Mn reconstituted rSODs and the insensitivity of the native SOD to both cyanide and H2O2. Both the native SOD and MnrSOD were determined to be homotetramers with monomeric molecular mass of 26 kDa and 27.5 kDa, respectively. They had high thermostability at 50° C and 60° C, and showed striking stability across a wide pH span from 4.0 to 11.0.

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Authors and Affiliations

  1. School of Biotechnology Engineering, Jimei University, Xiamen, 361021, China

    Yanbing Zhu, Xuqin Zhang, Chunyan Zhang, Jionghua Xiang & Guangming Liu

  2. 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. Xuqin Zhang
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  4. Chunyan Zhang
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  5. Jionghua Xiang
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  6. Guangming Liu
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Correspondence to Guangming Liu.

Additional information

Foundation item: The Natural Science Foundation of Fujian Province, China under contract Nos 2008J0067 and 2009J01033; the Program for New Century Excellent Talents in Fujian Province University under contract No. NCETFJ-2007; the Foundation for Innovative Research Team of Jimei University under contract No. 2010A005.

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Zhu, Y., Li, H., Zhang, X. et al. Characterization of a thermostable manganese-containing superoxide dismutase from inshore hot spring thermophile Thermus sp. JM1. Acta Oceanol. Sin. 30, 95–103 (2011). https://doi.org/10.1007/s13131-011-0166-8

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  • DOI: https://doi.org/10.1007/s13131-011-0166-8

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