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Unique Characteristics of Recombinant Hybrid Manganese Superoxide Dismutase from Staphylococcus equorum and S. saprophyticus

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Abstract

A recombinant hybrid of manganese dependent-superoxide dismutase of Staphylococcus equorum and S. saprophyticus has successfully been overexpressed in Escherichia coli BL21(DE3), purified, and characterized. The recombinant enzyme suffered from degradation and aggregation upon storage at −20 °C, but not at room temperature nor in cold. Chromatographic analysis in a size exclusion column suggested the occurrence of dimeric form, which has been reported to contribute in maintaining the stability of the enzyme. Effect of monovalent (Na+, K+), divalent (Ca2+, Mg2+), multivalent (Mn2+/4+, Zn2+/4+) cations and anions (Cl−, SO4 2−) to the enzyme stability or dimeric state depended on type of cation or anion, its concentration, and pH. However, tremendous effect was observed with 50 mM ZnSO4, in which thermostability of both the dimer and monomer was increased. Similar situation was not observed with MnSO4, and its presence was detrimental at 200 mM. Finally, chelating agent appeared to destabilize the dimer around neutral pH and dissociate it at basic pH. The monomer remained stable upon addition of ethylene diamine tetraacetic acid. Here we reported unique characteristics and stability of manganese dependent-superoxide dismutase from S. equorum/saprophyticus.

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Abbreviations

EDTA:

Ethylene diamine tetra acetate

LC–MS/MS:

Liquid chromatography tandem mass spectrometry

ORF:

Open reading frame

PAGE:

Polyacrylamide gel electrophoresis

rMnSODSeq:

Recombinant hybrid manganese superoxide dismutase from S. saprophyticus/S. equorum

SDS:

Sodium dodecyl sulphate

SO:

Superoxide

SOD:

Superoxide dismutase

T m :

Melting temperature

T mD :

Melting temperature of dimer

T mM :

Melting temperature of monomer

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Acknowledgments

This work is financially supported by Program Riset Desentralisasi DIKTI 2013 and Program Riset Inovasi 2015.

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Authors and Affiliations

  1. Laboratory of Pharmaceutical Biotechnology, School of Pharmacy, Bandung Institute of Technology, Jalan Ganesa No. 10, Bandung, 40132, Indonesia

    Debbie S. Retnoningrum, Anis Puji Rahayu, Dina Mulyanti & Astrid Dita

  2. Institute for Microbiology and Genetics, Georg-August-University, Grisebachstraße 8, 37077, Göttingen, Germany

    Oliver Valerius

  3. Dexa Laboratories of Biomolecular Sciences, Kawasan Industri Jababeka II, Jl. Industri Selatan V Blok PP No. 7, Cikarang, 17550, Indonesia

    Wangsa T. Ismaya

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  1. Debbie S. Retnoningrum
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Correspondence to Debbie S. Retnoningrum.

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Retnoningrum, D.S., Rahayu, A.P., Mulyanti, D. et al. Unique Characteristics of Recombinant Hybrid Manganese Superoxide Dismutase from Staphylococcus equorum and S. saprophyticus . Protein J 35, 136–144 (2016). https://doi.org/10.1007/s10930-016-9650-5

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