Purification and Characterization of Versatile Peroxidase from citrus sinensis Leaf Extract and its Application in Green Chemistry
Nivedita Rai, Meera Yadav, Hardeo Singh Yadav
Abstract
Nivedita Rai, Meera Yadav, Hardeo Singh Yadav
Abstract
Versatile peroxidase, a new family of ligninolytic peroxidases have catalytic properties of both lignin peroxidase and manganese peroxidase and has been isolated from citrus sinensis leaf extract which contains manganese peroxidase activity of 2.2 IU/ml and lignin peroxidase activity of 0.44 IU/ml. The overall recovery yield was 11 % with specific activity 1.0 IU/mg. Its molecular wt was found to be 24.60KDa. Km and Kcat value using veratryl alcohol and manganese sulphate as a substrate is 20 μM, 7.87sec and 12.5 μM, 13.84sec-1. The calculated pH optimum was 2.4 ± 0.1 for lignin peroxidase activity and 4.5 ± 0.1 for manganese peroxidase activity. The temperature optimum of the enzyme was 18°C for LiP and 25°C for MnP. Degradation of polyaromatic hydrocarbons like α-naphthol, β-naphthol, and 1,10-phenanthroline has been studied using UV/VIS spectrophotometer and cyclovoltameter at room temperature. Enzyme activity was inhibited by sodium azide and EDTA effectively.
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Versatile peroxidase, a new family of ligninolytic peroxidases have catalytic properties of both lignin peroxidase and manganese peroxidase and has been isolated from citrus sinensis leaf extract which contains manganese peroxidase activity of 2.2 IU/ml and lignin peroxidase activity of 0.44 IU/ml. The overall recovery yield was 11 % with specific activity 1.0 IU/mg. Its molecular wt was found to be 24.60KDa. Km and Kcat value using veratryl alcohol and manganese sulphate as a substrate is 20 μM, 7.87sec and 12.5 μM, 13.84sec-1. The calculated pH optimum was 2.4 ± 0.1 for lignin peroxidase activity and 4.5 ± 0.1 for manganese peroxidase activity. The temperature optimum of the enzyme was 18°C for LiP and 25°C for MnP. Degradation of polyaromatic hydrocarbons like α-naphthol, β-naphthol, and 1,10-phenanthroline has been studied using UV/VIS spectrophotometer and cyclovoltameter at room temperature. Enzyme activity was inhibited by sodium azide and EDTA effectively.
Key concepts: Chemistry, Peroxidase, Lignin peroxidase, Sodium azide, Manganese peroxidase, Manganese, Lignin, Nuclear chemistry