Liquid chromatographic and electrophoretic characterisation of extracellular beta-glucosidase of Pleurotus ostreatus grown in organic waste.
The production of beta-glucosidase by the ligninolytic fungus Pleurotus ostreatus has been studied in different culture media containing agro-industrial wastes. The enzyme is purified by anion-exchange chromatography, the molecular mass and isoelectric point of purified beta-glucosidase are measured by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing and the stability and kinetic parameters of the enzyme assessed by spectrophotometry. It has been established that the retention time, molecular mass and isoelectric point of the enzyme depend on the composition of the culture media while the activity and stability of beta-glucosidases of different origin were very similar. The combined chromatographic and electrophoretic methods have proved to be suitable techniques for the purification and characterisation of the beta-glucosidases produced by the ligninolytic fungus Pleurotus ostreatus in different culture media.
Morais
Helena
H
National Agronomical Station, Quinta do Marques, Oeiras, Portugal.
Ramos
Cristina
C
Matos
Natalia
N
Forgács
Esther
E
Cserháti
Tibor
T
Almeida
Vitor
V
Oliveira
José
J
Darwish
Yassin
Y
Ilés
Zoltán
Z
D000837:Anion Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007220:Industrial Waste; D007526:Isoelectric Point; D007700:Kinetics; D008970:Molecular Weight; D020076:Pleurotus / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet; D001617:beta-Glucosidase / Q000032:analysis* / Q000378:metabolism
D000837:Anion Exchange Resins; D007220:Industrial Waste; D001617:beta-Glucosidase