Recombinant vectors construction for cellobiohydrolase encoding gene constitutive expression

Abstract

Cellobiohydrolases (EC 3.2.1.91) are important exo enzymes involved in cellulose hydrolysis alongside endoglucanases (EC 3.2.1.4) and β- glucosidases (EC 3.2.1.21). Heterologous cellobiohydrolase gene expression under constitutive promoter control using Saccharomyces cerevisiae as host system is of great importance for a successful SSF process. From this point of view, the main objective of the work was to use Yeplac181 expression vector as a recipient for cellobiohdrolase - cbhB encoding gene expression under the control of the actin promoter, in Saccharomyces cerevisiae. Two hybrid vectors, YEplac-Actp and YEplac-Actp-CbhB, were generated using Escherichia coli XLI Blue for the cloning experiments. Constitutive cbhB gene expression was checked by proteine gel electrophoresis (SDS-PAGE) after insertion of these constructs into Saccharomyces cerevisiae.