Expression and characterisation of a pH and salt tolerant, thermostable and xylose tolerant recombinant GH43 β-xylosidase from Thermobifida halotolerans YIM 90462T for promoting hemicellulose degradation

Antonie Van Leeuwenhoek. 2019 Mar;112(3):339-350. doi: 10.1007/s10482-018-1161-2. Epub 2018 Sep 17.

Abstract

A gene encoding a β-xylosidase (designated as Thxyl43A) was cloned from strain Thermobifida halotolerans YIM 90462T. The open reading frame of this gene encodes 550 amino acid residues. The gene was over-expressed in Escherichia coli and the recombinant protein was purified. The monomeric Thxyl43A protein presented a molecular mass of 61.5 kDa. When p-nitrophenyl-β-d-xylopyranoside was used as the substrate, recombinant Thxyl43A exhibited optimal activity at 55 °C and pH 4.0 to 7.0, being thermostable by maintaining 47% of its activity after 30 h incubation at 55 °C. The recombinant enzyme retained more than 80% residual activity after incubation at pH range of 4.0 to 12.0 for 24 h, respectively, which indicated notable thermostability and pH stability of Thxyl43A. Moreover, Thxyl43A displayed high catalytic activity (> 60%) in presence of 5-35% NaCl (w/v) or 1-20% ionic liquid (w/v) or 1-50 mM xylose. These properties suggest that Thxyl43A has potential for promoting hemicellulose degradation and other industrial applications.

Keywords: Salt-tolerance; Thermobifida halotolerans YIM 90462T; Thermostability; Xylose-tolerance; pH-stability; β-xylosidase.

MeSH terms

  • Actinobacteria / enzymology*
  • Actinobacteria / genetics
  • Cloning, Molecular
  • Enzyme Stability
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Molecular Weight
  • Polysaccharides / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Sodium Chloride / metabolism
  • Xylosidases / chemistry
  • Xylosidases / genetics
  • Xylosidases / isolation & purification
  • Xylosidases / metabolism*

Substances

  • Polysaccharides
  • Recombinant Proteins
  • Sodium Chloride
  • hemicellulose
  • Xylosidases
  • exo-1,4-beta-D-xylosidase