title: Comparative purification and characterization of two distinct extracellular monocrotophos hydrolases secreted by Penicillium aculeatum and Fusarium pallidoroseum isolated from agricultural fields.
abstract: The present study aimed at a comparative characterization of two distinct extracellular monocrotophos hydrolases, from Penicillium aculeatum ITCC 7980.10 (M3) and Fusarium pallidoroseum ITCC 7785.10 (M4), isolated from agricultural fields. The MCP hydrolases were purified by Sephadex G-100 column and DEAE-Sepharose CL-6B ion-exchange column followed by SDS-PAGE analysis, which showed the presence of two hydrolases, of 33 and 67 kDa respectively. Both enzymes were most active at alkaline pH and were stable over a wide range of temperatures (60-70 °C). Between the strains, the MCP hydrolases from M3 were 2-fold more active than that from M4. Enzyme kinetic studies showed lowest Km (33.52 mM) and highest Vmax (5.18 U/mg protein) for OPH67 of M3 in comparison to the Km and Vmax of the other hydrolases purified from M3 and M4, suggesting that M3 OPH67 was the most efficient MCP hydrolase. To the best of our knowledge, this is the first report of the purification of two distinct extracellular thermostable MCP hydrolases from fungal strains Penicillium aculeatum ITCC 7980.10 and Fusarium pallidoroseum ITCC 7785.10. Owing to its potential MCP hydrolyzing activity, M3 OPH67 can perhaps used directly or in the encapsulated form for remediation of MCP contaminated sites.
authors.affiliation: Department of Bioscience and Biotechnology, Banasthali Vidyapith, Rajasthan, India.
authors.forename: Rachna
authors.initials: R
authors.lastname: Jain
authors.forename: Veena
authors.initials: V
authors.lastname: Garg
authors.forename: Koushalya
authors.initials: K
authors.lastname: Dangwal
authors.forename: Madhuri Kaushish
authors.initials: MK
authors.lastname: Lily
mesh_terms: D000383:Agriculture; D000577:Amides / Q000737:chemistry; D001673:Biodegradation, Environmental; D004795:Enzyme Stability; D005110:Extracellular Space / Q000201:enzymology*; D005670:Fusarium / Q000166:cytology* / Q000302:isolation & purification; D006867:Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008999:Monocrotophos / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010407:Penicillium / Q000166:cytology* / Q000302:isolation & purification; D010575:Pesticides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000577:Amides; D010575:Pesticides; D008999:Monocrotophos; D006867:Hydrolases