Purification, biochemical characterization, and structure of recombinant endo-1,4-ß-xylanase XylE.
Biochemistry (Mosc)
; 77(10): 1190-8, 2012 Oct.
Article
em En
| MEDLINE
| ID: mdl-23157299
ABSTRACT
The gene xylE encoding endo-1,4-ß-xylanase from the 10th family of glycosyl hydrolases produced by the mycelial fungus Penicillium canescens has been expressed under the control of the strong promoter of the bgaS gene encoding ß-galactosidase from P. canescens. As a result, a strain-producer of endoxylanase XylE was developed. The recombinant enzyme was isolated and purified to homogeneity with specific activity of 50 U/mg. The physicochemical and biochemical properties of the endoxylanase were studied. The maximal enzymatic activity was observed at pH 6.0 and 70°C. Endoxylanase XylE was shown to be a highly thermostable enzyme with half-inactivation period τ(1/2) of 7 h at 60°C. The kinetic parameters were 0.52 mg/ml (K(m)) and 75 µmol/min per mg (V(max)) using birch xylan as the substrate. Crystals of endoxylonase XylE were obtained, and the 3D structure was solved at 1.47 Å resolution. The 3D structure of an endo-1,4-ß-xylanase from the 10th family containing carbohydrate and unique cyclic structure located at the C-terminus of the polypeptide chain was obtained for the first time.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Penicillium
/
Proteínas Recombinantes
/
Proteínas de Escherichia coli
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Simportadores
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Endo-1,4-beta-Xilanases
Idioma:
En
Revista:
Biochemistry (Mosc)
Ano de publicação:
2012
Tipo de documento:
Article