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Characterization of a neutral pectin lyase produced by Oidiodendron echinulatum MTCC 1356 in solid state fermentation

Author:
Yadav, Sangeeta, Dubey, Amit Kumar, Anand, Gautam, Yadav, Dinesh
Source:
Journal of basic microbiology 2012 v.52 no.6 pp. 713-720
ISSN:
0233-111X
Subject:
Citrus, Crotalaria juncea, EDTA (chelating agent), activation energy, agricultural wastes, ammonium sulfate, anion exchange, denaturation, enzyme activity, enzyme inhibition, enzyme inhibitors, fungi, metal ions, pH, pectin lyase, pectins, polyacrylamide gel electrophoresis, potassium permanganate, retting, solid state fermentation, temperature, wheat bran
Abstract:
A neutral pectin lyase produced by a new fungal strain Oidiodendron echinulatum MTCC 1356 under solid state fermentation using wheat bran as agro waste has been studied. The enzyme was purified by ammonium sulphate precipitation (30–60%), DEAE anion exchange and Sephadex G‐100 column chromatographies. The SDS‐PAGE and native PAGE revealed two bands of sizes 42 and 47 kDa. The enzyme was purified 37 fold with specific activity of 4.5 U/mg and 2.25% yield. The Kₘ and Vₘₐₓ values determined using citrus pectin were 1.2 mg/ml and 0.36 IU/min respectively. The pH and temperature optima were pH 7.0 and 50 °C, respectively. The pH stability was around 5.0 for 24 h at 20 °C. The purified enzyme retained maximum activity for 30 min upto 50 °C. The activation energy for thermal denaturation of the purified enzyme was found to be 60.0 kJ/Mol. The effects of various metal ions and protein inhibitors on enzyme activity have revealed total inhibition of the enzyme activity in the presence of Ag⁺and Cu⁺and KMnO₄ at 1 mM. The neutral pectin lyase showed retting of Crotalaria juncea fibre in the presence of EDTA. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Agid:
204774