원문정보
초록
영어
Pseudomonas spp. have alginate biosynthetic gene cluster and produce bacterial alginates as 1-4-linked beta-D-mannuronan. We had isolated Pseudomonas fluorescens, Strain KS408 with poly-mannuronate lyase activity and identified the open reading frame of alginate lyase gene. In this study, the poly-mannuronate lyase gene was cloned to the expression vector of pET21b(+) vector and characterized. The alginate lyase of the Strain KS408 was composed of 1,110 nucleotides and 369 amino acids.
pET21b(+)/KS408 plasmid was transformed into E. coli, Strain BL21(DE3) and the protein of alginate lyase was expressed with IPTG induction. The alginate lyase fusion protein in the homogenate of recombinant E. coli was bound onto Ni-Sepharose affinity chromatography and eluted 300 mM imidazole solution. The purified enzyme was characterized the effects of temperature, pH and metal ions on the alginate lyase activity were investigated.