title: Purification and biochemical characterization of an alkaline pectin lyase from Fusarium decemcellulare MTCC 2079 suitable for Crotalaria juncea fiber retting.
abstract: An extracellular pectin lyase secreted by Fusarium decemcellulare MTCC 2079 under solid state fermentation condition has been purified to electrophoretic homogeniety by using ammonium sulfate fractionation, carboxymethyl cellulose and gel filtration (Sephadex G-100) column chromatographies. The purified enzyme showed single protein band corresponding to molecular mass 45 ± 01 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme had maximum activity at pH 9.0 and showed maximum stability in the pH range of 9.0-12.0. The optimum temperature of the purified enzyme was 50 °C and it showed maximum stability upto 40 °C. The energy of activation for the thermal denaturation (Ea ) was 59.06 kJ mol(-1)  K(-1). The Km and kcat values using citrus pectin as the substrate were 0.125 mg ml(-1) and 72.9 s(-1) in 100 mM sodium carbonate buffer pH 9.0 at 50 °C. The biophysical studies on pectin lyase showed that its secondary structure belongs to α + β class of protein with comparatively less of β-sheets. Purified pectin lyase showed efficient retting of Crotolaria juncea fibers.
authors.affiliation: Department of Biotechnology, DDU Gorakhpur University, Gorakhpur, India.
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Yadav
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Dubey
authors.forename: Gautam
authors.initials: G
authors.lastname: Anand
authors.forename: Reetesh
authors.initials: R
authors.lastname: Kumar
authors.forename: Dinesh
authors.initials: D
authors.lastname: Yadav
mesh_terms: D005591:Chemical Fractionation; D002853:Chromatography, Liquid; D029864:Crotalaria / Q000737:chemistry*; D004043:Dietary Fiber / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005670:Fusarium / Q000201:enzymology* / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011487:Protein Conformation; D013696:Temperature
chemical_list: D004043:Dietary Fiber; D011133:Polysaccharide-Lyases; C027349:pectin lyase
keywords: Cell wall degrading enzyme; Crotolaria juncea fiber; Fusarium decemcellulare; Pectin lyase; Pectinase; Retting; Tissue maceration