March 2018, Vol. 3, No. 3, pp 65-71.
Heterologous Expression of Exoglucanase from Trichoderma resei in E. coli
R. Navnit kumar¹, S. Jason Charles², T. R. Sambavi¹, S. Kabilan³, S. Renganathan¹,*
¹Biofuels Laboratory, Centre for Biotechnology, Anna University, Chennai. India.
²Molecular Biology Laboratory, Centre for Biotechnology, Anna University, Chennai. India.
³Bioprocess Laboratory, Centre for Biotechnology, Anna University, Chennai. India.
*Corresponding author’s e-mail: firstname.lastname@example.org
FPases/Exoglucanases are usually produced in lesser quantity from the wild cellulolytic strains in comparison to the quantities of Endoglucanases, Beta glucosidases and Xylanases. In the present work, an intracellular soluble expression of exoglucanase was attempted in a prokaryotic expression host E. coli SHuffle. A gene Cel7a that codes for Exgolucanases/FPases was isolated from Trichoderma resei and ligated into the pRSET B vector. E. coli SHuffle strain was transformed with the plasmid vector. A His-Tag metal affinity purification was performed after a fermentation that lasted for 3.5 hr using the recombinant E.coli Shuffle strain. Around 0.7 g/L of exoglucanases of 58 KDa molecular weight was obtained after the purification. The recombinant Exoglucanase had an activity of around 2.5 FPU/mL when assessed using the standard IUPAC Ghose assay for cellulase.
Keywords: Bioethanol; Fermentation; Cellulase; Vector; Prokaryotic expression.
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