Purification and characterization of an intracellular β-glucosidase from Botrytis cinerea
1995; Elsevier BV; Volume: 17; Issue: 10 Linguagem: Inglês
10.1016/0141-0229(94)00143-f
ISSN1879-0909
AutoresYannick Gueguen, Patrick Chemardin, A. Arnaud, P. Galzy,
Tópico(s)Polysaccharides and Plant Cell Walls
ResumoThe filamentous fungus Botrytis cinerea, grown on a cellobiose medium, was found to produce three intracellular β-glucosidases (GluI, II, and III). These enzymes were visualized by activity staining after separation by native polyacrylamide gel electrophoresis. GluI, which represents 95.5% of the total β-glucosidase activity, was purified to homogeneity by ion-exchange chromatography and gel filtration. The molecular mass of the purified intracellular β-glucosidase estimated by gel filtration was 350 kDa. The tetrameric structure of the β-glucosidase was determined following treatment of the purified enzyme with dodecyl sulphate. The intracellular β-glucosidase exhibited optimum catalytic activity at 50°C and pH 7 with citrate-phosphate buffer and 6.5 with phosphate buffer. The enzyme was active against glycosides with (1 → 4)-β, (1 → 2)-β, and (1 → 4)-α linkage configuration. The β-glucosidase was competitively inhibited by glucose and byd-gluconic-acid-lactone, and a glucosyl transferase activity was observed in the presence of ethanol.
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