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Isolation, Purification, Characterization of Cellulolytic Enzymes Produced by the Isolate Streptomyces omiyaensis

Author(s): M.Z. Alam | M.A. Manchur | M.N. Anwar

Journal: Pakistan Journal of Biological Sciences
ISSN 1028-8880

Volume: 7;
Issue: 10;
Start page: 1647;
Date: 2004;
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Keywords: cmc-ase | fp-ase | avecelase | b-glucosidase | sds-page | streptomyces omiyaensis

A Streptomyces strain A2 which was provisionally identified as Streptomyces omiyaensis, isolated from goat`s rumen was found capable of producing cellulolytic enzymes during growth on different cellulosic substrates. The isolate A2 grown under different conditions showed that CMC was the best cellulosic substrates for inducing the synthesis of extracellular cellulolytic enzymes. The isolate also showed heavy growth and liquefaction at pH 6.5, temperature 35 to 40�C and 5 days of incubation period during growth in liquid Winsted`s media having 1.2% CMC as a cellulose substrate. Maximum level of CMC-ase (230.56 U mL-1) in liquid media was found to produce when beef extract used as a nitrogen source along with CMC as cellulose source. Maximum level of reducing sugar (473.33 μg mL-1) and protein (324.27 μg mL-1) was obtained when CMC used as a growth substrate. The crude enzyme of the isolate was found to show highest enzyme activity (CMC-ase of 269.44 U mL-1, FP-ase of 64.81 U mL-1, Avicelase of 200 U mL-1 and β-Glucosidase of 138.89 U mL-1) at pH 6.5 and temperature 45�C during enzyme substrate reaction. The molecular weight of enzyme of the isolate Streptomyces omiyaensis was determined by SDS-PAGE technique and found 85 kDa.

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