Artigo Revisado por pares

Characterization of the xylanase produced by submerged cultivation of Thermomyces lanuginosus DSM 10635

2004; Elsevier BV; Volume: 35; Issue: 1 Linguagem: Inglês

10.1016/j.enzmictec.2004.04.003

ISSN

1879-0909

Autores

Hairong Xiong, Antti Nyyssölä, Janne Jänis, Ossi Pastinen, Niklas von Weymarn, Matti Leisola, Ossi Turunen,

Tópico(s)

Enzyme Catalysis and Immobilization

Resumo

Abstract Xylanase was produced by growing Thermomyces lanuginosus DSM 10635 in different carbohydrate containing media. The xylanase was purified to electrophoretic homogeneity and some of its enzymatic properties were studied. The molecular mass of this xylanase estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 25.5 kDa, and the p I was approximately 3.7. Applying mass spectrometric analysis, the average molecular mass of the xylanase was determined to be 21295.17 Da, which is practically the same as the theoretical molecular mass of the T. lanuginosus DSM 5826 xylanase (crystal structure 1yna). Accordingly, the thermostability and enzymatic properties of DSM 10635 xylanase were similar to DSM 5826 xylanase. The inactivation of DSM 10635 xylanase during the elevation of temperature proceeded slowly at pH 6.5 and 8.0, whereas at pH 5.0, the inactivation profile was steep, indicating that changes in the electrostatic interactions decrease the stability at lower pH. The thermostabilizing effect of the substrate on DSM 10635 xylanase was studied and it was observed to be significant only under acidic conditions. The half-life in the presence of 1% birchwood xylan (w/v) was 7-fold, 4-fold and 1.5-fold higher than without the substrate at pH 4.0, 5.0 and 6.5, respectively. No increase in the stability by the presence of substrate was observed at pH 9.0.

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