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

Ali Irtaza Rizvi; Ayesha Nawaz; Rabail Afzal; Muhammad Asim; Sayed Ali Hamza; Tazmeen Shahid; Urooj Rasheed; Maryam Manzoor; Nawal Batool; Zainab Aqsa; Nimra Hanif

doi:10.71336/jabs.1499

Authors

Ali Irtaza Rizvi University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0005-5242-212X
Ayesha Nawaz University of Central Punjab Lahore, Faculty of Science and Technology, Department of Microbiology, Lahore, Pakistan https://orcid.org/0009-0003-2329-1656
Rabail Afzal University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0009-7251-2842
Muhammad Asim University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0001-0997-7630
Sayed Ali Hamza University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0002-2399-3909
Tazmeen Shahid University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0009-8012-2692
Urooj Rasheed University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0007-2577-5282
Maryam Manzoor University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0008-7712-9706
Nawal Batool University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0002-3461-883X
Zainab Aqsa University of Central Punjab Lahore, Faculty of Science and Technology, Department of Food Science and Technology, Lahore, Pakistan https://orcid.org/0009-0005-7578-0405
Nimra Hanif University of Central Punjab Lahore, Faculty of Science and Technology, Department of Biotechnology, Lahore, Pakistan https://orcid.org/0009-0001-7869-380X

DOI:

https://doi.org/10.71336/jabs.1499

Keywords:

Xylanase gene, Escherichia coli BL21, Polymerase Chain Reaction, Bacillus cereus, pET 21c( )

Abstract

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.

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

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