Production of an oligosaccharide-specific cellobiohydrolase from the thermophilic fungus Thielavia terrestris

Biotechnol Lett. 2016 May;38(5):825-32. doi: 10.1007/s10529-016-2045-z. Epub 2016 Feb 1.


Objectives: To express and determine the hydrolytic activity of a cellobiohydrolase (TTCBH6B) from the thermophilic fungus Thielavia terrestris in Pichia pastoris.

Results: Ttcbh6B encodes a protein of 507 amino acid residues with a predicted molecular mass of 54 kDa. TTCBH6B contains a familial 6-glycosyl hydrolase catalytic domain and a type I carbohydrate-binding module. TTCBH6B was expressed and purified to homogeneity but the purified enzyme was inactive against Avicel. It could, however, digest Celluclast-treated Avicel producing cellobiose (0.27 μmol min(-1) mg(-1)). To determine the substrate preferences of TTCBH6B, oligosaccharides of varying numbers of subunits were generated by acid hydrolysis of Avicel and fluorescently tagged. Peaks corresponding to oligosaccharides containing three to six glucose units were reduced to cellobiose after addition of TTCBH6B.

Conclusion: TTCBH6B is active against shorter oligosaccharides rather than polysaccharides.

Keywords: 2-Aminobenzamide; Avicel; Carbohydrate-binding module; Cellobiose; Celluclast; Endoglucanases; Glycosyl hydrolases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cellobiose / metabolism
  • Cellulose 1,4-beta-Cellobiosidase / chemistry
  • Cellulose 1,4-beta-Cellobiosidase / genetics
  • Cellulose 1,4-beta-Cellobiosidase / isolation & purification
  • Cellulose 1,4-beta-Cellobiosidase / metabolism*
  • Molecular Weight
  • Oligosaccharides / metabolism*
  • Pichia / genetics
  • Pichia / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sordariales / enzymology*
  • Sordariales / genetics
  • Substrate Specificity


  • Oligosaccharides
  • Recombinant Proteins
  • Cellobiose
  • Cellulose 1,4-beta-Cellobiosidase