Molecular Cloning and Expression of Cellulase of Gene of Pseudomonas sp. in Escherichia coli

Pseudomonas sp.의 Cellulase 유전자의 대장균에의 클로닝 및 발현

  • 정영철 (경상대학교 식품공학과) ;
  • 김양우 (경상대학교 식품공학과) ;
  • 노종수 (경상대학교 식품공학과) ;
  • 성낙계 (경상대학교 식품공학과) ;
  • 강신권 (경상대학교 식품공학과)
  • Published : 1990.12.01

Abstract

The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

Keywords

Pseudomonas sp.;cellulase gene;cloning