EPrints@IIT Delhi >
Faculty Research Publicatons  >
Biochemical Engg. and Biotechnology >

Please use this identifier to cite or link to this item: http://eprint.iitd.ac.in/handle/2074/318

Title: Expression and characterization of Pichia etchellsii B-glucosidase in Escherichia coli
Authors: Pandey, Manjula
Mishra, Saroj
Keywords: Cellulose hydrolysis
Oligosaccharide biosynthesis
Yeast enzyme expression
Broad-substrate-specificity enzyme
Issue Date: 1997
Citation: Gene,190(1), 45-51
Abstract: The B-glucosidase enzyme is important as the terminal enzyme involved in hydrolysis of cellobiose and short-chain cellodextrins generated during enzymatic cellulose degradation. Under controlled reaction conditions the enzyme also displays cello-oligosaccha-ride synthesizing ability (based on either the thermodynamic or kinetic approach). We present here the purification of the enzyme P-glucosidase (BGL) of Pichia etchellsii from recombinant pBG55 Escherichia coli clone. The kinetic parameters, substrate specificity and oligosaccharide synthesizing ability of the purified enzyme are also reported. The purified 200-kDa protein (tetramer of 50 kDa) was identified as a broad-substrate-specificity enzyme exhibiting increased temperature and glucose tolerance compared to the native yeast enzyme. Temperature directed substrate specificity for aryl fl,l-4 linkage, and B( l-2), j3( l-4), fl( l-6) and 8(2-1) linkages in various natural disaccharides was observed. Glycosylation of the enzyme was found to be unimportant for enzyme activity. With both cellobiose and glucose, oligosaccharide synthesis was detected. The implications of this information with regard to cellulose hydrolysis and oligosaccharide synthesis are discussed.
URI: http://eprint.iitd.ac.in/dspace/handle/2074/318
Appears in Collections:Biochemical Engg. and Biotechnology

Files in This Item:

File Description SizeFormat
pandeyexp97.pdf707.56 kBAdobe PDFView/Open
View Statistics

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.


Valid XHTML 1.0! DSpace Software Copyright © 2002-2010  Duraspace - Feedback