Isolation and Characterization of Acid Protease Produced by Staphylococcus sp. CB2-3 from Digestive Organ of Harmonia axyridis

무당벌레 소화기관으로부터 산성 단백질 분해효소를 생산하는 Staphylococcus sp. CB2-3의 분리 및 특성

  • Kim, Se-Jong (Department of Microbial & Nano Materials, Mokwon University) ;
  • Whang, Kyung-Sook (Department of Microbial & Nano Materials, Mokwon University)
  • 김세종 (목원대학교 미생물나노소재학과) ;
  • 황경숙 (목원대학교 미생물나노소재학과)
  • Received : 2011.09.15
  • Accepted : 2011.09.28
  • Published : 2011.09.30

Abstract

Six protein-degrading bacteria were isolated from digestive organ of Harmonia axyridis. These isolates were categorized as Staphylococcus sciuri subsp. sciuri (3 strains), Bacillus subtilis (1 strain), and Bacillus thuringiensis (2 strains) by 16S rRNA gene sequence analysis. The Staphylococcus sp. CB2-3 was selected as a protease-producing bacterium which showed the highest protease activity of 58.5 U/ml at the pH 5.0 medium. The optimal pH and temperature of protease activity were pH 5.0 and $40^{\circ}C$, respectively. This acid protease had a relatively high stability of 80% between $30-50^{\circ}C$ at broad temperature range. The opimal medium compositions of carbon, nitrogen and mineral source for cell growth and protease activity were investigated. When sorbitol (0.5%) was used as carbon source, enzyme activity was increased about 2 times than that of the basal medium. When skim milk (0.5%) was used as nitrogen source, activity was increased about 2.5 times than that of the control. Cell growth and enzyme activity were increased by mineral source such as KCl, $K_2HPO_4$, $FeSO_4$, but was completely inhibited by divalent ions such as $Co^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Cu^{2+}$.

Keywords

Harmonia axyridis;Staphylococcus sp. CB2-3;acid protease;digestive organ

Acknowledgement

Supported by : 농촌진흥청

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