Skip to main content
SpringerLink
Log in

A highly stable Cu/Zn superoxide dismutase from Withania somnifera plant: gene cloning, expression and characterization of the recombinant protein

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

A gene from Withania somnifera (winter cherry), encoding a highly stable chloroplastic Cu/Zn superoxide dismutase (SOD), was cloned and expressed in Escherichia coli. The recombinant enzyme (specific activity of ~4,200 U mg−1) was purified and characterized. It retained ~90 and ~70% residual activities after 1 h at 80 and 95°C, respectively. At 95°C, thermal inactivation rate constant (K d) of the enzyme was 2.46 × 10−3 min−1 and half-life of heat inactivation was 4.68 h. The enzyme was stable against a broad pH range (2.5–11.0). It also showed a high degree of resistance to detergent, ethanol and protease digestion. This recombinant Cu/Zn SOD could therefore have useful applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bafana A, Dutt S, Kumar S, Ahuja PS (2010) Superoxide dismutase: an industrial perspective. Crit Rev Biotechnol 31:65–76

    Article  PubMed  Google Scholar 

  • Bannister JV, Bannister WH, Rotilio G (1987) Aspects of the structure, function, and applications of superoxide dismutase. CRC Crit Rev Biochem 22:111–180

    Article  PubMed  CAS  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287

    Article  PubMed  CAS  Google Scholar 

  • Chen C-N, Pan S-M (1996) Assay of superoxide dismutase activity by combining electrophoresis and densitometry. Bot Bull Acad Sin 37:107–111

    CAS  Google Scholar 

  • Kumar S, Sahoo R, Ahuja P (2002) Isozyme of autoclavable superoxide dismutase (SOD), a process for the identification and extraction of the SOD in cosmetic, food and pharmaceutical compositions. United States Patent 6,485,950

  • Lim JH, Yu YG, Han YS, Cho S, Ahn BY, Kim SH, Cho Y (1997) The crystal structure of fe-superoxide dismutase from the hyperthermophile Aquifex pyrophilus at 1.9 a resolution: structural basis for thermostability. J Mol Biol 270:259–274

    Article  PubMed  CAS  Google Scholar 

  • Lin CT, Lin MT, Chen YT, Shaw JF (1995) Subunit interaction enhances enzyme activity and stability of sweet potato cytosolic Cu/Zn-superoxide dismutase purified by a his-tagged recombinant protein method. Plant Mol Biol 28:303–311

    Article  PubMed  CAS  Google Scholar 

  • Mikael JL, Johanna N, Arne H, Mikael O (2004) Folding of human superoxide dismutase: disulfide reduction prevents dimerization and produces marginally stable monomers. Proc Natl Acad Sci USA 101:15893–15898

    Article  Google Scholar 

  • Parge HE, Hallewell RA, Tainer JA (1992) Atomic structures of wild-type and thermostable mutant recombinant human Cu, Zn superoxide dismutase. Proc Natl Acad Sci USA 89:6109–6113

    Article  PubMed  CAS  Google Scholar 

  • Prashanth SR, Sadhasivam V, Parida A (2008) Over expression of cytosolic copper/zinc superoxide dismutase from a mangrove plant Avicennia marina in Indica rice var Pusa Basmati-1 confers abiotic stress tolerance. Transgenic Res 17:281–291

    Article  PubMed  CAS  Google Scholar 

  • Tim DO, James DC, Zuzana V, Jan JE (1996) Human extracellular superoxide dismutase is a tetramer composed of two disulphide-linked dimers: a simplified, high-yield purification of extracellular superoxide dismutase. Biochem J 317:51–57

    Google Scholar 

  • Tuli R, Sangwan RS et al (2009) In: Tuli R, Sangwan RS (eds) A monograph, Aswagandha (Withania somnifera) a model Indian medicinal plant. CSIR, New Delhi, Sep 2009. ISBN no: 978-93-80235

  • Youn HD, Kim EJ, Roe JH, Hah YC, Kang SO (1996) A novel nickel-containing superoxide dismutase from streptomyces spp. Biochem J 318:889–896

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank the NMITLI programme of CSIR, India for promoting research on Withania somnifera, and NBRI, Lucknow, India for providing research facility. RT thanks the Department of Science and Technology for JC Bose Fellowship. VG, FD and SKU are thankful to CSIR for Senior Research Fellowship. Authors are grateful to Rajesh Srivastava for technical support and to Sarah Jamil and Saurabh Verma for metal analysis.

Author information

Authors and Affiliations

  1. National Botanical Research Institute, Council of Scientific and Industrial Research, Rana Pratap Marg, Lucknow, UP, 226001, India

    Raju Madanala, Vijayta Gupta, Farah Deeba, Santosh Kumar Upadhyay, Vivek Pandey, Pradhyumna Kumar Singh & Rakesh Tuli

  2. Department of Biotechnology, National Agri-Food Biotechnology Institute, C-127, Phase VIII, Industrial Area, SAS Nagar, Mohali, Punjab, 160071, India

    Rakesh Tuli

Authors
  1. Raju Madanala
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vijayta Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Farah Deeba
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Santosh Kumar Upadhyay
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vivek Pandey
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pradhyumna Kumar Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rakesh Tuli
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rakesh Tuli.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 31 kb)

Supplementary material 2 (DOCX 195 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Madanala, R., Gupta, V., Deeba, F. et al. A highly stable Cu/Zn superoxide dismutase from Withania somnifera plant: gene cloning, expression and characterization of the recombinant protein. Biotechnol Lett 33, 2057–2063 (2011). https://doi.org/10.1007/s10529-011-0670-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-011-0670-0

Keywords

Search

Navigation