Paper Title:
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.

  Info
Periodical
Chapter
Chapter 2: Future Materials Technology and Biomedicine
Edited by
Barry Tan
Pages
367-374
DOI
10.4028/www.scientific.net/AMR.365.367
Citation
H. Luo, Y. Zhou, Y. H. Chang, L. Xiong, L. Z. Liu, "Rapid Gene Cloning, Overexpression and Characterization of a Thermophilic Catalase in E. coli", Advanced Materials Research, Vol. 365, pp. 367-374, 2012
Online since
October 2011
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