| Title | Streptomyces griseus Trypsin (SGT) Has Gelatinase Activity and Its Proteolytic Activity Is Enhanced by Manganese |
|---|---|
| Author | Won-Jae Chi 1 , Yoon-Hee Kim 1 , Jong-Hee Kim 1 , Dae-Kyung Kang 2 , Sang-Soon Kang 3 , Joo-Won Suh 1 , and Soon-Kwang Hong 1 , * |
| Address | 1 Department of Biological Science, College of Natural Science, Myongji University, San 38-2,Nam-Dong, Yongin, Kyunggi-do 449-728, Korea; 2 Bio-Resources Institute, Easy Bio System Inc., Chonan, Chungnam 330-820, Korea ; 3 Division of Science Education, Chungbuk National University, Chongju 361-763, Korea |
| Bibliography | Journal of Microbiology, 41(4),289-294, 2003, |
| DOI | |
| Key Words | Streptomyces griseus Trypsin, SGT, serine protease, gelatinase |
| Abstract | Gelatinase is a proteolytic enzyme that hydrolyzes gelatin. Gelatinolytic activity was detected from culture broths of Streptomyces griseus IFO13350 and HH1 by paper disc assays on 0.5% agar plates containing 1% gelatin. The concentrated extracellular protein from the S. griseus was analyzed by SDS polyacrylamide gel, and two proteins, with molecular weights of 30 and 28 kDa, respectively, were identified to have gelatinase activity by gelatin zymography. The protein with a molecular weight of 28 kDa was confirmed to be S. griseus trypsin (SGT). The effects of metal ions and metal chelators on the protease activity of the SGT were studied. Of the metal ions tested, only manganese was found to enhance the protease activity, 2.6 times, however, Co_2^+, Cu_2^+, and Zn_2^+, and metal chelators, such as EDTA and EGTA, inhibited the SGT activity. When the protease activity of the SGT was measured at various pHs, in the presence of 5 mM MnCl_2, its highest activity was at pH 11.0, whereas only 60% of the maximum activity was observed between pHs 4.0 and pH 6.0, and almost 80% activity between pHs 7.0 to pH 10.0. The protease activity was measured at various temperatures in the presence of 5 mM MnCl_2. The SGT was found to be stable up to 60℃ for 30 min, while only 16% of the enzyme activity remained at 60℃, and at 80℃ almost all the activity was lost. The optimal temperature for the protease activity was 50℃. |
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