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Development of a Recombinant Streptomyces griseus with sprA and sprB Genes for Proteolytic Enzyme Production  

Hwang Ji-Hwan (Division of Food Science and Biotechnology, Kyungnam University)
Lee Chang-Kwon (Bio Food and Drug Research Center, Konkuk University)
Lee Kang-Mu (Division of Food Science and Biotechnology, Kyungnam University)
Jo Byoung-Kee (Division of Food Science and Biotechnology, Kyungnam University)
Park Hae-Ryong (Division of Food Science and Biotechnology, Kyungnam University)
Hwang Yong-Il (Division of Food Science and Biotechnology, Kyungnam University)
Publication Information
Korean Journal of Microbiology / v.41, no.1, 2005 , pp. 87-92 More about this Journal
Abstract
Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.
Keywords
integration vector; pronase; sprA; sprB; Streptomyces griseus;
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