Purification and characterization of pectin lyase secreted by Aspergillus flavus MTCC 10938.
An indigenously isolated fungal strain Aspergillus flavus MTCC 10938 was subjected to pectin lyase (PNL) production under submerged fermentation conditions. The enzyme was purified to homogeneity from the culture filtrate of the fungus involving concentration by ultrafiltration, anion exchange chromatography on DEAE cellulose and gel filtration chromatography on Sephadex G-100. The purified PNL gave a single protein band in SDS-PAGE analysis with a relative molecular mass corresponding to 50 kDa. Using citrus pectin as the substrate the K(m) and k(cat) values of the enzyme lyase were obtained as 1.7 mg/mL and 66 s(-1), respectively. The optimum pH of the purified PNL from A. flavus MTCC 10938 was 8.0 and up to 90% of its activity retained in the pH range from 3.0 to 11.0 after 24 h incubation. The optimum temperature of the purified enzyme was revealed at 55 degrees C and it was completely stable up to 40 degrees C when exposed for 30 min. The purified A. flavus MTCC 10938 PNL showed efficient retting of Crotalaria juncea fibres.
Department of Biotechnology, D.D.U. Gorakhpur University, Gorakhpur 273009, India. sangeeta_rahul@rediffmail.com
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Yadav
Department of Biotechnology, D.D.U. Gorakhpur University, Gorakhpur 273009, India.
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Dubey
Department of Biotechnology, D.D.U. Gorakhpur University, Gorakhpur 273009, India.
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Anand
Department of Biotechnology, D.D.U. Gorakhpur University, Gorakhpur 273009, India.
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Yadav
D001231:Aspergillus flavus / Q000201:enzymology* / Q000254:growth & development; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005285:Fermentation; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010368:Pectins / Q000378:metabolism*; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature
D005656:Fungal Proteins; D010368:Pectins; D011133:Polysaccharide-Lyases; C027349:pectin lyase