Mycobiology.  2009 Jun;37(2):121-127.

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

• Begum MF
• Absar N

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 45degrees C and the highest activity was exhibited in 35 to 45degrees C. The enzyme lost their activities almost completely (95~100%) at 80 degrees C or above and as well as bellow 25degrees C.

Keyword

Aspergillus; DEAE-cellulose chromatography; Gel filtration; Intracellular cellulase; SDS-PAGE

MeSH Terms

Acrylamide
Aspergillus
Aspergillus oryzae
Benzyl Alcohols
Cellulase
Cellulose
Chromatography, DEAE-Cellulose
Chromatography, Gel
Chromatography, Ion Exchange
DEAE-Cellulose
Electrophoresis, Polyacrylamide Gel
Glucosides
Hydrogen-Ion Concentration
Molecular Weight
Oryza
Acrylamide
Benzyl Alcohols
Cellulase
Cellulose
DEAE-Cellulose
Glucosides