Purification and properties of a carboxymethylcellulase from phytopathogenic fungus Macrophomina phaseolina.
From the culture filtrate of Macrophomina phaseolina, two forms of carboxymethylcellulase were separated by ion-exchange chromatography and designated as CMCase-I and CMCase-II. CMCase-I was purified following a four-step procedure involving gel filtration on Sephadex G-75, Con-A Sepharose 4B affinity chromatography, fast protein liquid chromatography on mono Q anion-exchanger and on Superose 12 gel filtration. The final preparation was homogeneous by SDS-PAGE, isoelectric focussing in thin layers of polyacrylamide gels and immunoelectrophoresis. The enzyme showed optimum activity at pH 5.5 and 65 degrees C, was stable to heating at 65 degrees C for 10 min, and retained 31% of original activity after heating at 80 degrees C for 10 min. The molecular weight of the enzyme was 3.5 x 10(4) Da. A Km of 0.25 mg/ml was determined using carboxymethyl-cellulose as the substrate.
Roy
U
U
Vora
V C
VC
D002266:Carboxymethylcellulose Sodium / Q000302:isolation & purification*; D007527:Isoenzymes / Q000302:isolation & purification*; D008747:Methylcellulose / Q000031:analogs & derivatives*; D003904:Mitosporic Fungi / Q000201:enzymology*
D007527:Isoenzymes; D008747:Methylcellulose; D002266:Carboxymethylcellulose Sodium