V.V. TUNGIKAR, A.E. GAWADE AND P.P. DIXIT
Abstract
Tyrosinase is a type of oxidoreductase enzyme which is found ubiquitously in all life form. There are various industrial applications of the tyrosinase enzyme such as production of melanin, LDihydroxyphenyl alanine (DOPA), cross linking agent, environmental cleanup. Due to structural complexity eukaryotic tyrosinases are unable to extract and studied. Bacterial tyrosinases are monomeric and can easily be purified. In the current study, the tyrosinase from the Brevundimonas diminuta PV 24 was partially purified by salting out, dialysis and size exclusion chromatographic technique. Enzyme was fractionated by 50% to 90% Ammonium sulphate saturation. The precipitate was dialyzed against 100 mM potassium phosphate buffer to remove the salts. Biegel P60 was used as gel matrix. A total of 32 elutions were collected and the enzyme was found to be in 6 to 12 elusions. The techniques enhanced the final purification up to 55 folds with increased activity from 45.03 U/ml to 149 U/ml. The stability of the enzyme was also determined and it concluded that the enzyme is stable at pH 7 and temperature 37 0C.