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Identification of Three Superoxide dismutase Genes from a Geobacillus sp.

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Abstract

We report the characterization of three Superoxide dismutase (sod) genes isolated from a bacterium in the Geobacillus genus. We isolated the bacterium from high-temperature pond mud and used 16S rRNA gene sequence to confirm its identity in the Geobacillus genus. The three genes Mn-sod, Fe/Mn-sod, and Cu/Zn-sod were cloned and analyzed. Their open reading frames are Mn-sod: 615 bp encoding a 204 amino acid protein; Fe/Mn-sod: 1,236 bp encoding a 411 amino acid protein; Cu/Zn-sod: 522 bp encoding a 173 amino acid protein. When these sod genes were expressed in Escherichia coli, only Mn-SOD was able to be purified. The activity of the purified Mn-SOD we got was about 2,730 U/mg. Studies of this Mn-SOD showed that it was thermostable at 60°, had 70% activity at 80° after 2.5 h, and still had 30% activity at 90° after 2.5 h. Mn-SOD activity required the ion Mn2+. Based on gel electrophoresis, we deduced that this Mn-SOD was a homotetramer. No activity was detected after the other two genes (Fe/Mn-sod, Cu/Zn-sod) were expressed in Escherichia coli, but activities were detected when expressed in Pichia pastoris.

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Abbreviations

sod/SOD:

Superoxide dismutase

PCR:

Polymerase chain reaction

RDB:

Regeneration dextrose base

BMGY:

Buffered glycerol complex

BMMY:

Buffered methanol complex

MD:

Minimal dextrose

MM:

Minimal methanol

LB:

Luria-Bertani

PAGE:

Polyacrylamide gel electrophoresis

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

MALDI-TOF:

Matrix-assisted laser desorption/ionization Time-of-flight mass spectrometry

Tris:

Trihydroxymethyl aminomethane

PDB code:

Protein dada bank code

NBT:

Nitro blue tetrazolium

References

  1. Arnold K, Bordoli L, Kopp J, Schwede T (2006) Bioinformatics 22:195–201

    Article  CAS  Google Scholar 

  2. Beauchamp C, Fridovich I (1971) Anal Biochem 44:276–287

    Article  CAS  Google Scholar 

  3. Bradford MM (1976) Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  4. Chang ST, Parker KN, Bauer MW, Kelly RM (2001) Methods Enzymol 330:260–269

    Article  CAS  Google Scholar 

  5. Gusakov AV, Sinitsyn AP, Salanovich TN, Bukhtojarov FE, Markov AV, Ustinov BB, van Zeijl C, Punt P, Burlingame R (2005) Enzyme Microb Technol 36:57–69

    Article  CAS  Google Scholar 

  6. He YZ, Fan KQ, Jia CJ, Wang ZJ, Pan WB, Huang L, Yang KQ, Dong ZY (2007) Appl Microbiol Biotechnol 75:367–376

    Article  CAS  Google Scholar 

  7. Hunter T, Bannister JV, Hunter GJ (2002) Eur Biochem 269:5137–5148

    Article  CAS  Google Scholar 

  8. Jackson SM, Cooper JB (1998) Biometals 11:159–173

    Article  CAS  Google Scholar 

  9. Keele BB, McCord JM, Fridovich I (1970) J Bio Chem 245:6176–6181

    CAS  Google Scholar 

  10. Kim EJ, Chung HJ, Suh B, Hah YC, Roe JH (1998) J Bacteriol 180:2014–2020

    CAS  Google Scholar 

  11. Li DC, Gao J, Li YL, Lu J (2005) Extremophiles 9:1–6

    Article  Google Scholar 

  12. Marklund S, Marklund G (1974) Eur J Biochem 47:469–474

    Article  CAS  Google Scholar 

  13. Meier B, Parak F, Desideri A, Rotilio G (1997) FEBS Lett 414:122–124

    Article  CAS  Google Scholar 

  14. Parker KN, Chhabra SR, Lam D, Callen W, Duffaud GD, Snead MA, Short JM, Mathur EJ, Kelly RM (2001) Biotechnol Bioeng 75:322–333

    Article  CAS  Google Scholar 

  15. Prabhakar R, Morokuma K, Musaev DG (2006) Comput Chem 27:1438–1445

    Article  CAS  Google Scholar 

  16. Saito N (1973) Arch Biochem Biophys 155:290–298

    Article  CAS  Google Scholar 

  17. Sambrook J, Russell DW (2002) In: Huang PT (ed) Molecular cloning: a laboratory manual. Science Press, Beijing, pp 1228–1232

  18. Schumann J, Wrba A, Jaenicke R, Stetter KO (1991) FEBS Lett 282:122–126

    Article  CAS  Google Scholar 

  19. Specka U, Mayer F, Antranikian G (1991) Appl Environ Microbiol 57:2317–2323

    CAS  Google Scholar 

  20. Tachibana Y, Fujiwara S, Takagi M, Imanaka T (1997) J Ferment Bioeng 83:540–548

    Article  CAS  Google Scholar 

  21. Takami H, Takaki Y, Chee GJ, Nishi S, Shimamura S, Suzuki H, Matsui S, Uchiyama I (2004) Nucleic Acids Res 32:6292–6303

    Article  CAS  Google Scholar 

  22. Van Der Maarel MJEC, Vos A, Sanders P, Dijkhuizen L (2003) Biocatal Biotransformation 21:199–208

    Article  Google Scholar 

  23. Youn HD, Kim EJ, Roe JH, Hah YC, Kang SO (1996) Biochem J 318:889–896

    CAS  Google Scholar 

  24. Zeigler DR (2001) In: Zeigler DR (ed) Catalog of strains, vol 3, 7th ed. Bacillus Genetic Stock Center, The Ohio State University, USA, pp7–14

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Authors and Affiliations

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, 100081, Beijing, China

    Yuanyuan Zou, Ling Yang, Guoxian Liu, Yinv Li & Zhifang Zhang

  2. Basic Medical and Biological Sciences College, Soochow University, No. 199 Wuzhong Renai Street, 215123, Suzhou, China

    Yuanyuan Zou & Yuexiong Zhu

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  1. Yuanyuan Zou
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  2. Ling Yang
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  3. Guoxian Liu
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  4. Yinv Li
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  5. Yuexiong Zhu
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  6. Zhifang Zhang
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Correspondence to Zhifang Zhang.

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Zou, Y., Yang, L., Liu, G. et al. Identification of Three Superoxide dismutase Genes from a Geobacillus sp.. Protein J 30, 66–71 (2011). https://doi.org/10.1007/s10930-010-9303-z

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