A method of purification, identification and characterization of β-glucosidase from Trichoderma koningii AS3.2774.

摘要: Abstract In this study, we used native gradient-polyacrylamide gel electrophoresis and electroelution (NGGEE) to purify enzymatic proteins from Trichoderma koningii AS3.2774. With method, purified eight classified them the cellulase system by comparing secretions of T. in inductive medium repressive medium. It resulted 24-fold β-glucosidase (BG) purification with a recovery rate 5.5%, specific activity 994.6 IU mg − 1 protein. The final yield BG reached 8 μg under purifying procedure NGGEE. We also identified using enzyme assay thin-layer chromatography MALDI-TOFMS. This had one subunit molecular mass 69.1 kDa as determined sodium dodecylsulfate-polyacrylamide electrophoresis. hydrolytic an optimal pH 5.0, temperature 50 °C, isoelectric point 5.68 K m for p-nitrophenyl-β- d -glucopyranoside 2.67 mM. Taken together, show that NGGEE is reliable method through which μg grade active can be purified.