Kinetics Study of Extracellular Detergent Stable Alkaline Protease from Rhizopus oryzae

Brazilian Archives of Biology and Technology

Zareena Mushtaq¹  Muhammad Irfan¹  Muhammad Nadeem¹  Mammona Naz¹  Quratulain Syed¹ 

关键词: Kinetics;    Purification;    Characterization;    Protease;    detergent stability;    Rhizopus oryzae;   

DOI  :  10.1590/S1516-8913201400071

来源: SciELO

【 摘 要 】

In this study, extracellular alkaline protease was produced from Rhizopus oryzae in submerged fermentation using dairy waste (whey) as a substrate. Fermentation kinetics was studied and various parameters were optimized. The strain produced maximum protease at initial medium pH of 6.0 medium depth of 26 mm, inoculum size of 2% at incubation temperature of 35ºC for 168 h of fermentation. Alkaline protease was purified to homogeneity by ammonium sulphate fractionation followed by sephadex G-100 chromatography. The molecular mass of alkaline protease was 69 kDa determined by 10% SDS-PAGE. The optimum pH and temperature of alkaline protease was 9.0 and 40ºC, respectively. Metal profile of the enzyme showed that the enzyme was non-metallic in nature. The Km , Kcat , Vmax and Kcat/Km values of purified protease were 7.0 mg/mL, 3.8 x102S-1, 54.30 µmol/min and 54.28 s-1mg -1.mL respectively, using casein as substrate. The purified alkaline protease had stability with commercial detergents.

【 授权许可】

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