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The journal of microbiology v.43 no.3, 2005년, pp.237 - 243  

Characterization of Calcium-Activated Bifunctional Peptidase of the Psychrotrophic Bacillus cereus

Kim Jong-Il    (Department of Food and Microbial Technology Seoul Women's University   ); Lee Sun-Min    (Department of Food and Microbial Technology Seoul Women's University   ); Jung Hyun-Joo    (Department of Food and Microbial Technology Seoul Women's University  );
  • 초록

    The protease purified from Bacillus cereus JH108 has the function of leucine specific endopeptidase. When measured by hydrolysis of synthetic substrate (N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide), the enzyme activity exhibited optimal activity at pH 9.0, $60^{\circ}C$ . The endopeptidase activity was stimulated by $Ca^{++},\;Co^{++},\;Mn^{++},\;Mg^{++},\;and\;Ni^{++}$ , and was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline, and EGTA. Addition of serine protease inhibitor, PMSF, resulted in the elimination of the activity. The endopeptidase activity was fully recovered from the inhibition of EDTA by the addition of 1 mM $Ca^{++}$ , and was partially restored by $Co^{++}\;and\;Mn^{++}$ , indicating that the enzyme was stabilized and activated by divalent cations and has a serine residue at the active site. Addition of $Ca^{++}$ increased the pH and heat stability of endopeptidase activity. These results show that endopeptidase requires calcium ions for activity and/or stability. A Lineweaver-Burk plot analysis indicated that the $K_m$ value of endopeptidase is 0.315 mM and $V_{max}$ is 0.222 ) is $0.222\;{\mu}mol$ of N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide per min. Bestatin was shown to act as a competitive inhibitor to the endopeptidase activity.


  • 주제어

    Bacillus cereus .   leucine endopeptidase.  

  • 참고문헌 (19)

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