Enzyme activity of a Phanerochaete chrysosporium cellobiohydrolase (CBHI.1) expressed as a heterologous protein from Escherichia coli
Author(s) -
R.L. Howard,
Peter Masoko,
E. Abotsi
Publication year - 2003
Publication title -
african journal of biotechnology
Language(s) - English
Resource type - Journals
ISSN - 1684-5315
DOI - 10.5897/ajb2003.000-1060
Subject(s) - phanerochaete , chrysosporium , enzyme , cellulase , chemistry , escherichia coli , heterologous , heterologous expression , biochemistry , microbiology and biotechnology , cellulose , clone (java method) , biology , gene , recombinant dna
The aim of this study was to produce a secreted, heterologously expressed Phanerochaete chrysosporium cellobiohydrolase (CBHI.1) protein that required no in vitro chemical refolding and to investigate the cellulolytic activity of the clone expressing the glutathione S-transferase (GST) fused CBHI.1 protein. Plate enzyme activity screening of E. coli cells transformed with pGEXcbhI.1 vector on carboxy-methyl-cellulose (CMC) produced several clones which produced clearing zones on CMC when induced. A randomly selected representative pGEXcbhI.1 clone produced hydrolysis on both Avicel and CMC when induced. Crude protein extracts obtained from the induced pGEXcbhI.1 clone exhibited time dependent enzymatic activity against both CMC and Avicel. Key words : Phanerochaete chrysosporium, cellobiohydrolase, cellulase activity, heterologous expression. African Journal of Biotechnology Vol.2(9) 2003: 296-300
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