Heterologous expression, purification and enzyme activity of Neocallimastix sp. CellulaseA (celA)

Abstract

The CellulaseA (celA) encoding gene was isolated from Neocallimastix sp. genomic DNA by PCR using gene specific primers and cloned into pRSETA vector, expressed in E.coli BL21DE3 as a soluble protein. The celA solubility was further improved by optimizing the IPTG concentration and induction temperature. The celA was purified by affinity chromatography using Ni-NTA resins and tested for its cellulolytic activity using CMC substrate by Zymogram. A band of ~47kDa corresponding to celA was observed in Zymogram