| ZHU Yanbing,LI Hebin,ZHANG Xuqin,ZHANG Chunyan,XIANG Jionghua,LIU Guangming. 2011. Characterization of a thermostable manganese-containing superoxide dismutase from inshore hot spring thermophile Thermus sp. JM1. Acta Oceanologica Sinica, (6):95－103 |
| Characterization of a thermostable manganese-containing superoxide dismutase from inshore hot spring thermophile Thermus sp. JM1 |
| Characterization of a thermostable manganese-containing superoxide dismutase from inshore hot spring thermophile Thermus sp. JM1 |
| Received:July 27, 2010 Revised:December 21, 2010 |
| DOI：10.1007/s13131-011-0166-8 |
| Key words:manganese superoxide dismutase thermostability purification reconstitution |
| 中文关键词: manganese superoxide dismutase thermostability purification reconstitution |
| 基金项目: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. |
| Author Name | Affiliation | E-mail | | ZHU Yanbing | School of Biotechnology Engineering, Jimei University, Xiamen 361021, China | | | LI Hebin | Xiamen Medical College, Xiamen 361008, China | | | ZHANG Xuqin | School of Biotechnology Engineering, Jimei University, Xiamen 361021, China | | | ZHANG Chunyan | School of Biotechnology Engineering, Jimei University, Xiamen 361021, China | | | XIANG Jionghua | School of Biotechnology Engineering, Jimei University, Xiamen 361021, China | | | LIU Guangming | School of Biotechnology Engineering, Jimei University, Xiamen 361021, China | jmswgc@hotmail.com |
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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℃ and 60℃, and showed striking stability across a wide pH span from 4.0 to 11.0. |
| 中文摘要: |
| 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℃ and 60℃, and showed striking stability across a wide pH span from 4.0 to 11.0. |
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