Enzyme activity of a Phanerochaete chrysosporium cellobiohydrolase (CBHI.1) expressed as a heterologous protein from Escherichia coli

  • RL Howard Microbiology, School of Molecular and Life Sciences, University of the North, P/Bag X1106, Sovenga, 0727, South Africa
  • P Masoko Microbiology, School of Molecular and Life Sciences, University of the North, P/Bag X1106, Sovenga, 0727, South Africa
  • E Abotsi Biochemistry, School of Molecular and Life Sciences, University of the North, P/Bag X1106, Sovenga, 0727, South Africa

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.

African Journal of Biotechnology Vol.2(9) 2003: 296-300

Author Biography

RL Howard, Microbiology, School of Molecular and Life Sciences, University of the North, P/Bag X1106, Sovenga, 0727, South Africa
Tel/fax +27 15 2682862
Published
2004-02-26
Section
Articles

eISSN: 1684-5315