World Journal of Microbiology and Biotechnology

, Volume 30, Issue 11, pp 2985–2993 | Cite as

Development of a cellulolytic Saccharomyces cerevisiae strain with enhanced cellobiohydrolase activity

  • Jiefang Hong
  • Huajun Yang
  • Kun Zhang
  • Cheng Liu
  • Shaolan Zou
  • Minhua Zhang
Original Paper


Consolidated bioprocessing (CBP) is a promising technology for lignocellulosic ethanol production, and the key is the engineering of a microorganism that can efficiently utilize cellulose. Development of Saccharomyces cerevisiae for CBP requires high level expression of cellulases, particularly cellobiohydrolases (CBH). In this study, to construct a CBP-enabling yeast with enhanced CBH activity, three cassettes containing constitutively expressed CBH-encoding genes (cbh1 from Aspergillus aculeatus, cbh1 and cbh2 from Trichoderma reesei) were constructed. T. reesei eg2, A. aculeatus bgl1, and the three CBH-encoding genes were then sequentially integrated into the S. cerevisiae W303-1A chromosome via δ-sequence-mediated integration. The resultant strains W1, W2, and W3, expressing uni-, bi-, and trifunctional cellulases, respectively, exhibited corresponding cellulase activities. Furthermore, both the activities and glucose producing activity ascended. The growth test on cellulose containing plates indicated that CBH was a necessary component for successful utilization of crystalline cellulose. The three recombinant strains and the control strains W303-1A and AADY were evaluated in acid- and alkali-pretreated corncob containing media with 5 FPU exogenous cellulase/g biomass loading. The highest ethanol titer (g/l) within 7 days was 5.92 ± 0.51, 18.60 ± 0.81, 28.20 ± 0.84, 1.40 ± 0.12, and 2.12 ± 0.35, respectively. Compared with the control strains, W3 efficiently fermented pretreated corncob to ethanol. To our knowledge, this is the first study aimed at creating cellulolytic yeast with enhanced CBH activity by integrating three types of CBH-encoding gene with a strong constitutive promoter Ptpi.


Endoglucanase β-Glucosidase Cellobiohydrolase Consolidated bioprocessing Yeast 

Supplementary material

11274_2014_1726_MOESM1_ESM.docx (42 kb)
Supplementary material 1 (DOCX 41 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Jiefang Hong
    • 1
    • 2
  • Huajun Yang
    • 1
    • 2
  • Kun Zhang
    • 1
  • Cheng Liu
    • 1
  • Shaolan Zou
    • 1
  • Minhua Zhang
    • 1
  1. 1.Tianjin R&D Center for Petrochemical TechnologyTianjin UniversityTianjinChina
  2. 2.School of Chemical Engineering and TechnologyTianjin UniversityTianjinChina

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