[논문]Characterization of Calcium-Activated Bifunctional Peptidase of the Psychrotrophic Bacillus cereus

Kim Jong-Il; Lee Sun-Min; Jung Hyun-Joo

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

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

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)

Abstract ▼ AI-Helper

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.

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문제 정의

This study describes the characterization of an endopep tidase from the psychrotrophic strain of Bacillus cereus. The results of the study revealed that the enzyme was optimally active at an alkaline pH of 8 - 9 and at 60℃.

제안 방법

However, the extracellular pro tease from Bacillus amyloliquefaciens demonstrated activity only with endopeptidase (Son and Kim, 2003). Enzymological properties of the enzyme, previously char acterized as leucine aminopeptidase, were analyzed in this study.
In this study, the characteristics of the leucine endopep tidase, stimulated by Ca++, from psychrotrophic Bacillus cereus were analyzed.

성능/효과

This study describes the characterization of an endopep tidase from the psychrotrophic strain of Bacillus cereus. The results of the study revealed that the enzyme was optimally active at an alkaline pH of 8 - 9 and at 60℃. On the basis of its optimum pH, this protease would be grouped into the alkaline protease category.

참고문헌 (19)

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