Enzyme activity of a Phanerochaete chrysosporium cellobiohydrolase (CBHI.1) expressed as a heterologous protein from Escherichia coli
Keywords:
Phanerochaete chrysosporium, cellobiohydrolase, cellulase activity, heterologous expressionAbstract
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.
