Applied Microbiology and Biotechnology

, Volume 100, Issue 11, pp 4959–4968 | Cite as

Comparison of catalytic properties of multiple β-glucosidases of Trichoderma reesei

  • Boyang Guo
  • Nobuaki Sato
  • Peter Biely
  • Yoshihiko Amano
  • Kouichi Nozaki
Biotechnologically relevant enzymes and proteins


Ten putative Trichoderma reesei β-glucosidase (BGL) isozymes were heterologously expressed in Escherichia coli and Aspergillus oryzae and purified to homogeneity. Catalytic properties of nine enzymes which showed hydrolytic activity on cellobiose and p-nitrophenyl-β-D-glucopyranoside (pNPG) were investigated. Three BGLs, encoded by the genes cel3A, cel3B, and cel3E, contained a predicted signal peptide, showed higher hydrolytic activity on cello-oligosaccharides than on pNPG, and preferred longer oligosaccharides. Another three putative extracellular BGLs, Cel3B, Cel3F, and Cel3G, and two intracellular enzymes, Cel3C and Cel3D, exhibited preference for pNPG. Intracellular Cel1A showed the highest affinity for cellobiose as a typical cellobiase. Four BGLs, Cel3A, Cel3B, Cel3E, Cel1A, that showed high activity against cello-oligosaccharides were capable of catalyzing transglycosylation reactions from cellobiose, leading to formation of cellotriose and isomeric glucobioses. While Cel3A, Cel3B, and Cel3E synthesized mainly gentiobiose, glycosyl transfer reactions of Cel1A led mainly to sophorose and laminaribiose. Conversion of cellobiose to sophorose by Cel1A reached about 3.6 and 10 % at 1 and 10 % cellobiose concentration, respectively. The formation and persistence of individual cellobiose isomers in incubation mixtures of four BGLs (Cel3A, Cel3B, Cel3E, and Cel1A) with cellobiose correlated well with the kcat values for isomeric glucobioses. Cel1A also showed the lowest sensitivity to inhibition by glucose. Based on all studied catalytic properties, Cel1A appears to be unambiguously the best candidate for site-directed mutations or directed evolution toward improvement of activity, thermostability, and, eventually, efficiency of sophorose synthesis.


β-glucosidase Trichoderma reesei Transglycosylation Cellulose degradation 

Supplementary material

253_2016_7342_MOESM1_ESM.pdf (331 kb)
ESM 1(PDF 331 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Boyang Guo
    • 1
  • Nobuaki Sato
    • 1
  • Peter Biely
    • 2
  • Yoshihiko Amano
    • 1
  • Kouichi Nozaki
    • 1
  1. 1.Department of Bioscience and Textile Technology, Interdisciplinary Graduate School of Science and TechnologyShinshu UniversityNaganoJapan
  2. 2.Institute of ChemistrySlovak Academy of SciencesBratislavaSlovakia

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