title: Unique Characteristics of Recombinant Hybrid Manganese Superoxide Dismutase from Staphylococcus equorum and S. saprophyticus.
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 (Ca(2+), Mg(2+)), multivalent (Mn(2+/4+), Zn(2+/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.
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, School of Pharmacy, Bandung Institute of Technology, Jalan Ganesa No. 10, Bandung, 40132, Indonesia. retnoningrum@indo.net.id.
authors.forename: Debbie S
authors.initials: DS
authors.lastname: Retnoningrum
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, School of Pharmacy, Bandung Institute of Technology, Jalan Ganesa No. 10, Bandung, 40132, Indonesia.
authors.forename: Anis Puji
authors.initials: AP
authors.lastname: Rahayu
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, School of Pharmacy, Bandung Institute of Technology, Jalan Ganesa No. 10, Bandung, 40132, Indonesia.
authors.forename: Dina
authors.initials: D
authors.lastname: Mulyanti
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, School of Pharmacy, Bandung Institute of Technology, Jalan Ganesa No. 10, Bandung, 40132, Indonesia.
authors.forename: Astrid
authors.initials: A
authors.lastname: Dita
authors.affiliation: Institute for Microbiology and Genetics, Georg-August-University, Grisebachstraße 8, 37077, Göttingen, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Valerius
authors.affiliation: Dexa Laboratories of Biomolecular Sciences, Kawasan Industri Jababeka II, Jl. Industri Selatan V Blok PP No. 7, Cikarang, 17550, Indonesia.
authors.forename: Wangsa T
authors.initials: WT
authors.lastname: Ismaya
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013210:Staphylococcus / Q000201:enzymology / Q000235:genetics*; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase
keywords: Dimer; Manganese dependent-superoxide dismutase; Monomer; Recombinant hybrid protein; Staphylococcus equorum; Thermal stability