ISSN 2756-3413
African Journal of Virology Research ISSN 2756-3413 Vol. 14 (4), pp. 001-004, April, 2020. © International Scholars Journals
Full Length Research Paper
Refolding and characterisation of a heterologous expressed Phanerochaete chrysosporium cellobiohydrolase (CBHI.2)
Howard RL
Microbiology, School of Molecular and Life Sciences, University of Limpopo, P/Bag X1106, Sovenga, 0727, South Africa. E-mail: howardr@ul.ac.za. Tel/fax: +27 15 268 2862.
Accepted 17 September, 2019
Abstract
Cloned Phanerochaete chrysosporium ME446 cbhI.2 cDNA was successfully expressed in Escherichia coli as an insoluble, internal, biologically inactive protein. In vitro chemical refolding restored the activity of the crude CBHI.2. However, this enzyme was active against 4-methylumbelliferyl-ß-D-cellobioside (MUC) and 4-methyllumbelliferyl-ß -D- lactopyranoside (MUL) substrates only. The crude enzyme lost almost 50% of its activity at 5 min at 100oC heat treatment whereas total inactivation was achieved at 30 min.
Key words: Phanerochaete chrysosporium, CBHI.2, chemical refolding, heat-inactivation.