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M.Tech / M.E / PhD Thesis | Biology | India | Volume 3 Issue 6, June 2014
Isolation; Screening and Purification of Cellulase from Cellulase Producing Klebsiella variicola RBEB3 (KF036184.1)
Gopinath.S.M [3] | Ismail Shareef [4] | Ashalatha [6] | Shreya Ranjit [2]
Abstract: The cellulase producing bacteria were isolated from soil samples of paper industry waste; cloth industry waste; kitchen waste; garden; earthworm. Isolates were characterized on the basis of their morphological; biochemical and molecular characteristics. Total 32 isolates were obtained by primary screening technique from which 19 isolates showed cellulolytic activity; 06 isolates showed maximum cellulase activity and 13 showed a minimum cellulase activity. The Potential isolates were obtained from kitchen waste soil and garden soil and by characteristic study was found to be Klebsiella variicola RBEB3 and the sequence submitted to NCBI with accession number KF036184.1. Further purification of the cellulase enzyme was carried out by ammonium sulphate precipitation followed by gel filtration chromatography. The molecular weight of the enzyme was found to be 45 kDa by SDS PAGE method. On performing zymogram staining; the cellulolytic activity was confirmed; the enzyme activity was found to be 0.092 IU/ml and protein concentration was found to be 4.8 mg/ml. The use of microorganisms offers a promising approach for the large scale production of enzymes and as a food supplement or in pharmaceutical industry.
Keywords: Cellulase producing bacteria, purification, cellulase, Klebsiella variicola, Ammonium sulphate precipitation, Gel filtration chromatography
Edition: Volume 3 Issue 6, June 2014,
Pages: 1398 - 1403