Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Biochemical and Genetic Characterization of Arazyme, an Extracellular Metalloprotease Produced from Serratia proteamaculans HY-3

  • Kwak, Jang-Yul (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Ki-Eun (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Shin, Dong-Ha (Insect BioTech Co., Ltd.) ;
  • Maeng, Jin-Soo (Laboratory of Biophysical Chemistry, National Heart, Lung and Blood Institute, National Institutes of Health) ;
  • Park, Doo-Sang (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Oh, Hyun-Woo (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Son, Kwang-Hee (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Bae, Kyung-Sook (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Ho-Yong (Insect Resources Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2007.05.31
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Abstract

Serratia proteamaculans HY-3 isolated from the digestive tract of a spider produces an extracellular protease named arazyme, with an estimated molecular mass of 51.5 kDa. The purified enzyme was characterized as having high activities at wide pH and temperature ranges. We further characterized biochemical features of the enzymatic reactions under various reaction conditions. The protease efficiently hydrolyzed a broad range of protein substrates including albumin, keratin, and collagen. The dependence of enzymatic activities on the presence of metal ions such as calcium and zinc indicated that the enzyme is a metalloprotease, together with the previous observation that the proteolytic activity of the enzyme was not inhibited by aspartate, cysteine, or serine protease inhibitors, but strongly inhibited by 1,10-phenanthroline and EDTA. The araA gene encoding the exoprotease was isolated as a 5.6 kb BamHI fragment after PCR amplification using degenerate primers and subsequent Southern hybridization. The nucleotide sequence revealed that the deduced amino acid sequences shared extensive similarity with those of the serralysin family of metalloproteases from other enteric bacteria. A gene(inh) encoding a putative protease inhibitor was also identified immediately adjacent to the araA structural gene.

Keywords

References

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