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Production and Characterization of Thermostable Protease from Bacillus licheniformis Isolated from Korean Traditional Soybean Paste

재래식 된장에서 분리된 Bacillus licheniformis의 내열성 Protease 특성과 생산성

  • Bae, Young Eun (Department of Food Science & Biotechnology, Woosong University) ;
  • Yoon, Ki-Hong (Department of Food Science & Biotechnology, Woosong University)
  • 배영은 (우송대학교 식품생물과학과) ;
  • 윤기홍 (우송대학교 식품생물과학과)
  • Received : 2012.10.16
  • Accepted : 2012.11.16
  • Published : 2012.12.31

Abstract

Among 63 Bacillus strains grown at $60^{\circ}C$ from sixteen samples of homemade Korean soybean paste, one strain was selected for producing the thermostable protease. The isolate has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. Culture filtrate of the isolate showed maximal protease activity at the reaction condition of $60-65^{\circ}C$ and pH 11. The culture filtrate retained more than 87% of initial protease activity after incubation for 30 min at $60^{\circ}C$ without substrate. In order to develop the medium composition, effects of ingredients including nitrogen sources, carbon sources, metal ions and phosphate were examined for protease production of the isolate. Lactose and soytone peptone were the most effective carbon and nitrogen source for the enzyme production. After the late logarithmic growth phase the isolate began to produce the protease, and the maximum protease productivity was reached to 550 unit/ml in the optimized medium consisting of lactose (3%), soytone peptone (1.5%), $MgSO_4$ (0.1%), $K_2HPO_4$ (0.03%), and $KH_2PO_4$ (0.03%) at 28 h of incubation.

Keywords

Bacillus licheniformis;properties;protease productivity;soybean paste

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