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http://dx.doi.org/10.4489/MYCO.2009.37.2.121

Purification and Characterization of Intracellular Cellulase from Aspergillus oryzae ITCC-4857.01  

Begum, Ferdousi (Department of Botany, University of Rajshahi)
Absar, Nurul (Department of Biochemistry, University of Rajshahi)
Publication Information
Mycobiology / v.37, no.2, 2009 , pp. 121-127 More about this Journal
Abstract
Purification and characterization of intracellular cellulase produced by A. oryzae ITCC-4857.01 are reported. The enzyme was purified by ion-exchange chromatography using DEAE-cellulose followed by Gel filtration. The purification achieved was 41 fold from the crude extract with yield of 27%. The purified enzyme showed single band on poly acrylamide gel. The molecular weight as determined by SDS-PAGE and gel filtration was 38 KDa and 38.6 KDa respectively and contained only one subunit. The enzyme is glycoprotien as nature and contained 0.67% neutral sugar. The apparent Km value of the enzyme against cellulose was 0.83%. The enzyme showed the highest relative ativities on CMC followed by avicel, salicin and filter paper. The optimum pH of activity was 5.5 and very slight activity was observed at or above pH 7.5 as well as bellow pH 3.5. The optimum tempreture of the activity was $45^{\circ}C$ and the highest activity was exhibited in 35 to $45^{\circ}C$. The enzyme lost their activities almost completely (95${\sim}$100%) at $80^{\circ}C$ or above and as well as bellow $25^{\circ}C$.
Keywords
Aspergillus; DEAE-cellulose chromatography; Gel filtration; Intracellular cellulase; SDS-PAGE;
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