Rapid Gene Cloning, Overexpression and Characterization of a Thermophilic Catalase in E. coli

Abstract:

A thermophilic catalase-encoding gene was rapidly obtained by means a PCR-based protocol with the genomic DNA mixture from compost culture as the template. The open reading frame of this gene is composed of 2208 base pairs, sharing 92.5% homology with the reported Bacillus stearothermophilus gene (NCBI genbank accession No. AB020234. 1). A prokaryotic expression plasmid pET-CATHis was constructed for the gene expression, and two recombinant E. coli, BL21(DE3)/pET-CATHis and BL21(DE3)pLysS/pET-CATHis were finally obtained. After culture optimization, the highest activities for these two strains in shaking flask culture were 74.3 U/ml and 1055.3 U/ml, respectively. The 6 His-tagged recombinant catalase was then purified by using immobilized metal affinity chromatography, and the properties of the purified protein were finally characterized.

• Periodical: Advanced Materials Research (Volume 365)
• Pages: 367-374
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.365.367
• Online since: October 2011
• Authors: Hui Luo, Yao Zhou, Yan Hong Chang, Lian Xiong, Lin Zhi Liu
• Keywords: Catalase, Enzyme Properties, Overexpression, Screening, Thermophilic

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