Production, characterization, cloning and sequence analysis of a monofunctional catalase from Serratia marcescens SYBC08.

摘要: A monofunctional catalase from Serratia marcescens SYBC08 produced by liquid state fermentation in 7 liter fermenter was isolated and purified ammonium sulfate precipitation (ASP), ion exchange chromatography (IEC), gel filtration (GF) characterized. Its sequence analyzed LC-MS/MS technique gene cloning. The highest production (20,289 U · ml(-1)) achieved after incubation for 40 h. had an estimated molecular mass of 230 kDa, consisting four identical subunits 58 kDa. High specific activity the (199,584 mg(-1) protein) 3.44 times higher than that Halomonas sp. Sk1 (57,900 protein). enzyme without peroxidase found to be atypical electronic spectrum catalase. apparent K(m) V(max) were 78 mM 188, 212 per µM H(2) O(2) heme(-1) s(-1), respectivly. displayed a broad pH range (pH 5.0-11.0), with optimal 7.0-9.0: It most active at 20 °C 78% 0 °C. thermo stability slightly compared commercial bovine liver. analysis confirmed deduced amino acid cloning SYBC08. 23 related catalases. Although site residues, NADPH-binding proximal residues heme, distal heme interacting water molecule well conserved catalases, weakly found. closely catalases pathogenic bacterium family Enterobacteriaceae. This result imply high plays significant role preventing those microorganisms Enterobacteriaceae against hydrogen peroxide resulted cellular damage. Calalase yield has potential industrial application scavenging peroxide.