Molecular cloning and expression of Bacillus cereus xylanase gene in E. coli bl21

Recombinant production of bacterial xylanases offers a cost-effective and industrially relevant alternative to native enzyme extraction. In this study, the xylanase gene from Bacillus Cereus (~1071 bp) was amplified by PCR and cloned into the pET-21c(+) expression vector (~5441 bp) for heterologous expression in Escherichia coli BL21. Ligation and transformation were confirmed through colony PCR and restriction digestion with BamHI, producing a distinct band of 6512 bp, consistent with the recombinant plasmid. The successful cloning and verification demonstrate the feasibility of generating functional recombinant xylanase in E. coli. Bacterial xylanases, unlike fungal enzymes, exhibit high stability across a broad pH range and elevated temperatures, enhancing their suitability for industrial applications such as pulp and paper processing, food, and biofuel production. This work establishes a foundation for large-scale recombinant xylanase production and provides a platform for further optimization of fermentation conditions to improve enzyme yield. Future studies will focus on optimizing expression levels, scaling up production, and evaluating the enzyme’s performance under industrial process conditions.