Applied Microbiology and Biotechnology

, Volume 45, Issue 3, pp 338–341

Construction of an Aspergillus nidulans multicopy transformant for the xlnB gene and its use in purifying the minor X24 xylanase

  • M. T. Fernández-Espinar
  • S. Vallés
  • F. Piñaga
  • J. A. Pérez-González
  • D. Ramón
SHORT CONTRIBUTION

DOI: 10.1007/s002530050693

Cite this article as:
Fernández-Espinar, M., Vallés, S., Piñaga, F. et al. Appl Microbiol Biotechnol (1996) 45: 338. doi:10.1007/s002530050693

Abstract

 Using recombinant DNA techniques, an Aspergillus nidulans multicopy transformant for the gene xlnB coding for the minor X24 xylanase has been constructed. When grown on glucose as sole carbon source this transformant secretes 114 U of xylanase (mg protein)-1. In this culture condition, X24 is the only xylanase secreted and the predominant protein in the culture filtrate. This strategy has been used to purify the X24 enzyme to homogeneity. The purified xylanase showed a single band on sodium dodecyl sulphate/ polyacrylamide gel electrophoresis with a molecular mass of 24 kDa and had an isoelectric point of approximately 3.5. The enzyme was a non-debranching endo-1,4-β-xylan xylanohydrolase highly specific for xylans and showed optimal activity at pH 5.5 and 52°C. The X24 xylanase had a Michaelis constant, Km, of 12.43 mg oat spelt xylan ml-1 and a Vmax of 1639 μmol min-1 (mg protein)-1.

Copyright information

© Springer-Verlag Berlin Heidelberg 1996

Authors and Affiliations

  • M. T. Fernández-Espinar
    • 1
  • S. Vallés
    • 1
  • F. Piñaga
    • 1
  • J. A. Pérez-González
    • 1
  • D. Ramón
    • 1
  1. 1.Departamento de Biotecnología de Alimentos, Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Cientificas, Apartado de Correos 73; 46100-Burjassot; Valencia, Spain. Fax: 34-6-3636301 e-mail: DRAMON@IATA.CSIC.ESES