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Purification, characterisation and application of α‐l‐rhamnosidase from Penicillium citrinum MTCC‐8897

Yadav, Sarita, Yadav, Vinita, Yadav, Sudha, Yadav, Kapil D.S.
International journal of food science & technology 2012 v.47 no.2 pp. 290-298
Penicillium citrinum, ammonium sulfate, carboxymethylcellulose, chromatography, culture filtrates, enzymes, hesperidin, molecular weight, naringin, odors, pH, polyacrylamide gel electrophoresis, proteins, rutin, sodium sulfate, temperature, wines
An α‐l‐rhamnosidase secreted by Penicillium citrinum MTCC‐8897 has been purified to homogeneity from the culture filtrate of the fungal strain using ammonium sulphate precipitation and cation‐exchange chromatography on carboxymethyl cellulose. The sodium dodecyl sulphate/polyacrylamide gel electrophoresis analysis of the purified enzyme gave a single protein band corresponding to the molecular mass 51.0 kDa. The native polyacrylamide gel electrophoresis also gave a single protein band confirming the enzyme purity. The Km and Vmax values of the enzyme for p‐nitrophenyl α‐l‐rhamnopyranoside were 0.36 mm and 22.54 μmole min−1 mg−1, respectively, and kcat value was 17.1 s−1 giving kcat/Km value of 4.75 × 104 m−1 s−1. The pH and temperature optima of the enzyme were 7.0 and 60 °C, respectively. The purified enzyme liberated l‐rhamnose from naringin, rutin, hesperidin and wine, indicating that it has biotechnological application potential for the preparation of l‐rhamnose and other pharmaceutically important compounds from natural glycosides containing terminal α‐l‐rhamnose and also in the enhancement of wine aroma.