ISSN 2756-3707
International Journal of Histology and Cytology ISSN 2447-9535 Vol. 2 (5), pp. 168-172, May, 2015. © International Scholars Journals
Full Length Research Paper
A study of secreted, heterologously expressed Phanerochaete chrysosporium cellobiohydrolase
Howard R.L.1*, Masoko P.1 and Abotsi E.2
1Microbiology, School of Molecular and Life Sciences, University of the North, P/Bag X1106, Sovenga, 0727, South Africa
2Biochemistry, School of Molecular and Life Sciences, University of the North, P/Bag X1106, Sovenga, 0727, South Africa.
Corresponding author: E-mail: [email protected], Tel/fax +27 15 2682862.
Accepted 02 March, 2014
Abstract
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.