Characterization and purification of thermostable beta-glucosidase from Clostridium thermocellum. |
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Authors: | N Aït N Creuzet J Cattanéo |
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Institution: | Laboratoire de Chimie Bactérienne, C.N.R.S. 13274 Marseille Cedex 2 France |
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Abstract: | A β-glucosidase was isolated from ; the enzyme was localized in the periplasmic space.It was purified in a five-step procedure including ion-exchange chromatography on DEAE-Cellulose, chromatography on HA-Ultrogel and DEAE-Sephadex, gel filtration on AcA 34 Ultrogel and isoelectric focusing.The final preparation was purified 944-fold with a recovery of about 5% of the initial enzyme activity.Polyacrylamide disc electrophoresis of the purified enzyme gave a single band at pH 8.3. The enzyme is active towards cellobiose and p-nitrophenyl-β-D-glucoside(PNPG) and developed maximum activities at pH 6.0 and 65°C. A molecular weight of 50,000 daltons was estimated by gel filtration and the enzyme was isoelectric at pH 4.68. |
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