Isolation and characterization of a thermostable endo-β-glucanase active on 1,3-1,4-β-D-glucans from the aerobic fungus Talaromyces emersonii CBS 814.70

Patrick G. Murray, Alice Grassick, Christopher D. Laffey, Michelle M. Cuffe, Timothy Higgins, Angela V. Savage, Antoni Planas, Maria G. Tuohy

Research output: Indexed journal article Articlepeer-review

89 Citations (Scopus)

Abstract

A novel endoglucanase active on 1,3-1,4-β-d-glucans was purified to apparent homogeneity from submerged cultures of the moderately thermophilic aerobic fungus Talaromyces emersonii CBS 814.70. The enzyme is a single subunit glycoprotein with Mr and pI values of 40.7 ± 0.3 kDa and 4.4, respectively, and an estimated carbohydrate content of 77% (w/w). The purified β-glucanase displayed activity over broad ranges of pH and temperature, yielding respective optima values of pH 4.8 and 80°C. This enzyme was markedly thermostable with 15% of the original activity remaining after incubation for 15 min at 100°C. Substrate specificity studies revealed the identity of the enzyme to be a 1,3-1,4-β-d-glucanase. Identical Km values (13.38 mg.ml-1) were obtained with lichenan and BBG, while the Vmax value with lichenan (142.9 IU.mg-1) was approximately twice the value obtained with BBG (79.3 IU.mg-1). Time-course hydrolysis of barley-β-glucan did not proceed linearly with respect to time indicating an 'endo' or more processive action for the enzyme. HPAEC fractionation of the products of hydrolysis yielded a range of oligosaccharides, with cellobiose, cellotriose and cellotetraose being the predominant oligosaccharide products.

Original languageEnglish
Pages (from-to)90-98
Number of pages9
JournalEnzyme and Microbial Technology
Volume29
Issue number1
DOIs
Publication statusPublished - 5 Jul 2001

Keywords

  • Barley-β-glucan
  • Glycoprotein
  • Lichenan
  • Talaromyces emersonii
  • Thermostable
  • β-glucanase

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