KSBB Journal

  • 제29권3호
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  • Pages.131-138
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  • 2014
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  • 1225-7117(pISSN)
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  • 2288-8268(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
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황토로부터 분리한 Bacillus licheniformis의 항진균 chitinase 생산과 효소 특성

Production and Characterization of Antifungal Chitinase of Bacillus licheniformis Isolated from Yellow Loess

  • 한귀환 (조선대학교 공과대학 환경공학과) ;
  • 봉기문 (조선대학교 공과대학 환경공학과) ;
  • 김종민 (조선대학교 공과대학 환경공학과) ;
  • 김평일 (전남생물산업진흥원 생물방제연구원) ;
  • 김시욱 (조선대학교 공과대학 환경공학과)
  • Han, Gui Hwan (Department of Environmental Engineering, Chosun University) ;
  • Bong, Ki Moon (Department of Environmental Engineering, Chosun University) ;
  • Kim, Jong Min (Department of Environmental Engineering, Chosun University) ;
  • Kim, Pyoung Il (Biol Control Research Institute) ;
  • Kim, Si Wouk (Department of Environmental Engineering, Chosun University)
  • 투고 : 2014.04.19
  • 심사 : 2014.05.08
  • 발행 : 2014.06.30
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초록

In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

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