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

Streptomyces griseus IFO13350 유래 sprA 및 sprB 유전자를 이용한 Pretense 생산균주 개발

  • 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)
  • 황지환 (경남대학교 식품생명공학부) ;
  • 이창권 (건국대학교 바이오 식의학 연구센터) ;
  • 이강무 (경남대학교 식품생명공학부) ;
  • 조병기 (경남대학교 식품생명공학부) ;
  • 박해룡 (경남대학교 식품생명공학부) ;
  • 황용일 (경남대학교 식품생명공학부)
  • Published : 2005.03.01

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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