title: Fusion of the OsmC domain from esterase EstO confers thermolability to the cold-active xylanase Xyn8 from Pseudoalteromonas arctica.
abstract: The OsmC-region (osmotically induced protein family) of the two-domain esterase EstO from the psychrotolerant bacterium Pseudoalteromonas arctica has been shown to increase thermolability. In an attempt to test if these properties can be conferred to another enzyme, we genetically fused osmC to the 3'-region of the family 8 xylanase encoding gene xyn8 from P. arctica. The chimeric open reading frame xyn8-OsmC was cloned and the chimeric protein was purified after heterologous expression in Escherichia coli. Xyn8 and Xyn8-OsmC showed cold-adapted properties (more than 60% activity at 0°C) using birchwood xylan as the preferred substrate. Maximal catalytic activity is slightly shifted from 15°C (Xyn8) to 20°C for Xyn8-OsmC. Thermostability of Xyn8-OsmC is significantly changed in comparison to wild-type Xyn8. The OsmC-fusion variant showed an apparent decrease in thermostability between 40 and 45°C, while both proteins are highly instable at 50°C.
authors.affiliation: Institute of Technical Microbiology, Hamburg University of Technology, Kasernenstr. 12, 21073 Hamburg, Germany.
authors.forename: Skander
authors.initials: S
authors.lastname: Elleuche
authors.forename: Henning
authors.initials: H
authors.lastname: Piascheck
authors.forename: Garabed
authors.initials: G
authors.lastname: Antranikian
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D043182:Carboxylesterase / Q000737:chemistry*; D002384:Catalysis; D003001:Cloning, Molecular; D003080:Cold Temperature; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D017434:Protein Structure, Tertiary; D031161:Pseudoalteromonas / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D043325:Xylan Endo-1,3-beta-Xylosidase / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; C099394:osmC protein, E coli; D043182:Carboxylesterase; D043325:Xylan Endo-1,3-beta-Xylosidase; D043364:Endo-1,4-beta Xylanases