title: Purification and characterization of thermoalkalophilic xylanase isolated from the Enterobacter sp. MTCC 5112.
abstract: Thermoalkalophilic Enterobacter sp. MTCC 5112 was isolated from a sediment sample collected from the Mandovi estuary on the west coast of India. This culture produced extracellular xylanase. The xylanase enzyme was isolated by ammonium sulfate (80%) fractionation and purified to homogeneity using size exclusion and ion exchange chromatography. The molecular mass of the xylanase was approximately 43 kDa. The optimal pH of the xylanase activity was 9, and at room temperature it showed 100% stability at pH 7, 8 and 9 for 3 h. The optimal temperature for the enzyme activity was 100 degrees C at pH 9.0. At 80 degrees C and pH 9, 90% of the enzyme activity was retained after 40 min. At 70 and 60 degrees C, the enzyme retained 64% and 85% of its activity after 18 h, respectively, while at 50 degrees C and pH 9 the enzyme remained stable for days. For xylan, the enzyme gave a K(m) value of 3.3 mg ml(-1) and a V(max) value of 5,000 micromol min(-1) mg(-1) when the reaction was carried out at 100 degrees C and pH 9. In the presence of metal ions such as Co(2+), Zn(2+), Fe(2+), Cu(2+), Mg(2+) and Ca(2+) the activity of the enzyme increased, whereas strong inhibition of enzyme activity was observed in the presence of Hg(2+) and EDTA. To the best of our knowledge, this is the first report on the production of xylanase by this bacterium.
authors.affiliation: National Institute of Oceanography, Dona Paula 403004, Goa, India. rakhee@darya.nio.org
authors.forename: Rakhee
authors.initials: R
authors.lastname: Khandeparkar
authors.forename: Narayan B
authors.initials: NB
authors.lastname: Bhosle
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002412:Cations / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry; D004492:Edetic Acid / Q000737:chemistry; D004754:Enterobacter / Q000201:enzymology*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008670:Metals / Q000737:chemistry; D013696:Temperature; D014995:Xylosidases / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D002412:Cations; D002614:Chelating Agents; D008670:Metals; D004492:Edetic Acid; D014995:Xylosidases