Extraction, Purification and Characterization of Antioxidant Enzyme Catalase (CAT) Present in Chili (Capsicum annuum, Var. Azad Mirch-1) Plant
2020
Katiyar, Saumya | Kumar, Anil
In this study, extraction, partial purification and characterization of an antioxidant enzyme catalase present in chilli (Capsicum annuum, var. Azad mirch-1) plant. Catalase enzyme was collected from fresh leaves, fruits and roots of chilli plant in the Rama University UP, Kanpur (India). 100 g of fresh tissues were taken and grinded in the mortar pistil till the fine homogenous slurry formed. Then 10g of PVPP was added along with 50 ml buffer (Sodium phosphate, 0.1M, pH 7.2), and 1g ascorbic acid. The extracted protein was fractionated with ammonium sulphate and partially purified with the use of SG-150 gel filtration column. Chilli leaf catalase has shown its optimum activity in pH 7.2 at 30oC. The native molecular weight of catalase present in the chilli leaf was found 59 kDa. The Km value was recorded 9.2 mM for H2O2 and Vmax value was found 1.41 IU/mg X 10-3 for the chilli leaf catalase. The enzyme has shown linearity after 8mg/ml enzyme concentration. Chlilli catalase was purified up to 50.83 purification fold. The catalase enzyme activity was also recorded in the chilli fruits and roots separately.
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