Activity Aspergillus oryzae to cellulase production | ||
| journal of kerbala university | ||
| Article 1, Volume 10, Issue 0, January 2014, Pages 15-27 PDF (0 K) | ||
| Authors | ||
| Athraa H. Muhsin; Mushin H. Resin; Fatima abide AL-Hussein | ||
| Abstract | ||
| This study aimed to extractioncellulase enzymes and purificatied by using many methods The crude enzymes when using ammonium sulfate had total activity (14 , 90 , 2835 )U, total protein 10.57 mg and the specific activity ( 0.013 , 1.57, 2.68 ) U/mg protein The crude culture filtrate containing Filter paperase (FPase) , Carboxymetheylcellulase (CMCase) and β-glucosidase were subjected to ammonium sulfate precipitation. Different concentrations (20, 40,60, 80 and 100%) of ammonium sulfate were used for that purpose. After conducting some preliminary trials, 40- 60 % concentrationwere selected for the enzyme precipitation. On the other hand,the protein of crude enzyme extract was separated in (33 ) fractions of fungi from DEAE-cellulose column is ,It was found that only the fractions ( F-8) contained cellulase activity., while other fractions were eluted by the buffer containing different concentrations of Nacl .On the other hand,the component of the active fraction ( F-8) , obtained after DEAE cellulose chromatography was separated by gel filtration and gave only one peak of enzymes alone and the fraction also found to be contained cellulase activity. as given in the specific activity of cellulase were ( 15.67 , 7.94 , 1972 .16 ) U/mg for FPase ,CMCase and β-glucosidase respectably and characterized purified enzyme effect of various pH values the highest enzyme activity was found at 5.5 pH. When the pH of the enzyme medium was increased up to 6.0 , FPase enzyme giving activity ( 0.91 ) U , but CMCase and β-glucosidase giving highest activityat pH 6 were ( 0.34 , 65.12 ) U and the activity declining at pH 5 to ( 0.25 , 59.95 ) IU . and It found that the enzyme exhibited maximum activity at 25 C° in CMCase was ( 0.33 ) U , but 30 C° for β-glucosidase was ( 38.22 ) U . On the other hand, FPase was having activity at 25 and 30 C° were (0.14,0.15)U. Finally,the crude enzyme preparation was subjected to SDS-PAGE to determine the molecular weight of the enzyme.While conducting the electrophoresis, one major band showing cellulolytic activity. The molecular weight of the protein was found to be about (38) kD, (52) kD and (49) kD for (FPase ,CMCase , β-glucosidase ) respectively. | ||
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