Molecular Cloning of Chitosanase Gene and Quantitative Production of Chitosan Oligomer

키토사네이즈 유전자의 클로닝과 키토산 올리고머의 정량적 생산

  • 박유미 (경북대학교 생명공학부) ;
  • 장혜란 (경북대학교 생명공학부) ;
  • 허태린 (경북대학교 생명공학부) ;
  • 김사열 (경북대학교 생명공학부)
  • Published : 2004.03.01

Abstract

Six bacterial strains which formed large halo on chitosan-containing agar plate were isolated from beach mud and crabs at South coast of Korean peninsula. They were designated as Bacillus cereus KNUC51, B. cereus KNUC52, B. cereus KNUC53, B. cereus KNUC54, B. cereus KNUC55, and Paenibacillus favisporus KNUC56 by analysing their morphologies and 16S rDNA sequences. Chitosanase activities of all isolates were similar to that of B. subtilis 168. To enhance the activity of chitosanase, a powerful mutagen, MNNG was treated for P favisporus KNUC56. Three mutants showed higher activity of chitosanase than that of the original strain. The DNA fragments containing chitosanase gene from B. cereus sources were cloned, sequenced, and their deduced amino acid sequence analysis showed over 93% homologies with that of the known B. cereus ATCC14579. Extracellular sample from the isolates was incubated in proper reaction mixture including chitosan for 5 minutes at $37^{\circ}C$ to produce 3-10 chitosan oligomers which has been known to be active for clinical agents and agronomical agents.

Keywords

Chitosan;16S rDNA;chitosanase;csn

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