1tux
HIGH RESOLUTION CRYSTAL STRUCTURE OF A THERMOSTABLE XYLANASE FROM THERMOASCUS AURANTIACUS
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OverviewOverview
Thermoascus aurantiacus xylanase is a thermostable enzyme which hydrolyses, xylan, a major hemicellulose component in the biosphere. Crystals, belonging to P21 space group with a=41.7 A, b=68.1 A, c=51. 4 A and, beta=113.6 degrees, Z=2 were grown that could diffract to better than 1.8, A resolution. The structure was solved by molecular replacement method, using the Streptomyces lividans xylanase model. The amino acid sequence, was determined from the electron density map aided by multiple alignment, of related xylanase sequences. The sequence thus obtained provides a, correction to the sequence reported earlier based on biochemical methods., The final refined protein model at 1.8 A resolution with 301 amino acid, residues and 266 water molecules has an R-factor of 16.0 % and free R of, 21.1 % with good stereochemistry. The single polypeptide chain assumes, (alpha/beta)8 TIM-barrel fold and belongs to F/10 family of glycoside, hydrolases. The active site consists of two glutamate residues located at, the C terminus end of the beta-barrel, conforming to the double, displacement mechanism for the enzyme action. A disulphide bond and more, than ten salt bridges have been identified. In particular, the salt bridge, Arg124-Glu232 which is almost buried, bridges the beta-strands beta4 and, beta7 where the catalytic glutamate residues reside, and it may play a key, role in the stability and activity at elevated temperature. To our, knowledge, for the first time in the F/10 family xylanases, we observe a, proline residue in the middle of the alpha-helix alpha6 which may be, contributing to better packing. Earlier studies show that the enzyme, retains its activity even at 70 degrees C. The refined protein model has, allowed a detailed comparison with the other known structures in the F/10, family of enzymes. The possible causative factors for thermostability are, discussed.
About this StructureAbout this Structure
1TUX is a Single protein structure of sequence from Thermoascus aurantiacus. Active as Endo-1,4-beta-xylanase, with EC number 3.2.1.8 Full crystallographic information is available from OCA.
ReferenceReference
Crystal structure at 1.8 A resolution and proposed amino acid sequence of a thermostable xylanase from Thermoascus aurantiacus., Natesh R, Bhanumoorthy P, Vithayathil PJ, Sekar K, Ramakumar S, Viswamitra MA, J Mol Biol. 1999 May 21;288(5):999-1012. PMID:10329194
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