STUDY ON THE PROPERTIES OF PURIFIED RECOMBINANT SUPEROXIDE DISMUTASE FROM STAPHYLOCOCCUS EQUORUM, A LOCAL ISOLATE FROM INDONESIA
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2014, Vol 6, Issue 11
Abstract
Objective: Superoxide dismutase (SOD) (E. C: 1.15.1.1) from Staphylococcus equorum which catalyzes the dismutation of the superoxide anion (O2.-) into molecular oxygen (O2) and hydrogen peroxide (H2O2), is one of the most important classes of antioxidant enzymes and are used in pharmaceutical or cosmetic applications. SOD of S. equorum was purified from total protein into homogeneity and characterized to determine the unit activity, ion metal cofactor, optimum temperature and pH, kinetic parameters, and effect of denaturing and reducing agents and UVC exposure on the rSOD activity.Methods: The protein was purified in a single-step purification using Ni-NTA afï¬nity column with various imidazole concentrations. SOD activity was analyzed by colorimetric and activity staining using nitroblue tetrazolium (NBT). The purified rSOD was exposed to different temperatures and pHs, different concentrations of denaturing agents, reducing agents, and to UVC exposure.Results: SOD protein with high purity was obtained when imidazole concentrations of 100 mM, 200 mM and 250 mM were applied. The purified rSOD displayed specific activity of 1666.7 U mg-1when measured at 30ºC and pH 7.8. The presence of conserved manganese-binding sites (H28, H83, D171, H175) and the inhibition of rSOD activity by NaN3 but not by H2O2 or KCN and indicated that rSOD was Mn-dependent. The optimum temperature and pH were determined to be 40ºC and 6.0, respectively. The Michaelis constant (Km), maximum velocity (Vmax), turnover number (kcat) and catalytic efficiency (kcat/Km) were found to be 371.2 µM, 1.738 µMS-1, 1.358 s-1, and 3.7x10-3 S-1µM-1, respectively. The rSOD activity was slightly affected in the presence of detergents (0.5% SDS, 0.5% Triton-X 100), denaturing agents (6 M GdnHCl and 6 M urea) and reducing agent (5 mM βME). After exposure of rSOD by UVC for 45 min, it retained half of its activity.Conclusion: This is the first study to report the stability of the SOD of S. equorum against environmental factors. The SOD displays some thermostability, is active in wide pH, stable in the presence of denaturing and reducing agents, however it is relatively unstable to UVC exposure
Authors and Affiliations
Ana Indrayati, Sukmadjaja Asyarie, Tri Suciati, Debbie Soefie Retnoningrum
Keywords
Related Articles
- EP ID EP579337
- DOI -
- Views 74
- Downloads 0
A STUDY ON EFFECT OF INDOLE AS A SUBSTITUENT ON A KETO-ENOL TAUTOMER: A SYNTHETIC APPROACH ON β-DIKETONE
Objective: The existence of keto-enol tautomerism in β-diketones can typically study by a choice of analytical technique. The position of the keto-enol equilibrium depends on a number of factors like solvent, temperatur...
THE MODULATING EFFECT OF BARLEY β-GLUCAN EXTRACT ON HIGH FAT DIET INDUCED OBESITY-A BIOCHEMICAL STUDY IN ALBINO WISTAR RATS
Objective: Diet-inducedhyperlipidemia and obesity are the major risk factors for type II diabetes mellitus, hypertension, musculoskeletal and cardiovascular disorders (CVD). The objective of the present study is to furni...
ENCAPSULATION OF ZIDOVUDINE IN DIFFERENT CELLULOSIC ACRYLIC AND METHACRYLIC POLYMERS LOADED MICROSPHERES: IN VITRO CHARACTERIZATION AND COMPATIBILITY STUDIES
Objective: The attempt of the present study was to improve bioavailability and dissolution rate along with reduction in dosing frequency of Zidovudine from microspheres.Methods: In this study an effort was taken to devis...
RESIDUAL VOLUMES AND FINAL WEIGHTS IN DIFFERENT TYPES OF PLASTIC INFUSION CONTAINERS
Objective: To compare the residual volumes, the drained volumes, and the final weights of different infusion containers with different volumes and degrees of flexibility. The risk of drug error can be aggravated by a hig...
PHARMACOGNOSTICAL, PHYTOCHEMICAL SCREENING AND STANDARDIZATION OF NELUMBO NUCIFERA (GAERTN): THE SACRED LOTUS
Objective: To perform the Phytochemical screening & standardization of Nelumbo nucifera Gaertn.Methods: The present study provides pharmacognostic, physicochemical and phytochemicals details of Parts of Nelumbo nucif...