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

ZHU Yanbing; LI Hebin; ZHANG Xuqin; ZHANG Chunyan; XIANG Jionghua; LIU Guangming

doi:10.1007/s13131-011-0166-8

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

doi: 10.1007/s13131-011-0166-8

1.
School of Biotechnology Engineering, Jimei University, Xiamen 361021, China
2.
Xiamen Medical College, Xiamen 361008, China

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.

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出版历程
- 收稿日期: 2010-07-27
- 修回日期: 2010-12-21

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

1.
School of Biotechnology Engineering, Jimei University, Xiamen 361021, China
2.
Xiamen Medical College, Xiamen 361008, China

摘要

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.
- manganese /
- superoxidedismutase /
- thermostability /
- purification /
- reconstitution
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.
- manganese /
- superoxide dismutase /
- thermostability /
- purification /
- reconstitution

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文章访问数: 285
HTML全文浏览量: 44
PDF下载量: 126
被引次数: 0

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

doi: 10.1007/s13131-011-0166-8

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

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出版历程

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