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A versatile Trichoderma reesei expression system for the production of heterologous proteins

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Abstract

Objectives

To develop a versatile Trichoderma reesei (teleomorph Hypocrea jecorina) expression system for the high-purity production of heterologous proteins.

Results

The versatile T. reesei expression system is based on xyn1 and xyn2 promoters, A824V transition in XYRI, and a bicomponent carbon source strategy. Red fluorescent protein gene rfp and alkaline endoglucanase EGV gene egv3 from Humicola insolens were used as reporter genes to test our versatile expression system

Conclusions

The versatile T. reesei expression system can be applied to produce heterologous proteins with high purity and high yield.

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References

  • Bischof RH, Ramoni J, Seiboth B (2016) Cellulases and beyond: the first 70 years of the enzyme producer Trichoderma reesei. Microb Cell Fact 15:106

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Cherry JR, Fidantsef AL (2003) Directed evolution of industrial enzymes: an update. Curr Opin Biotech 14:438–443

    Article  PubMed  CAS  Google Scholar 

  • Derntl C, Gudynaite-Savitch L, Calixte S, White T, Mach RL, Mach-Aigner AR (2013) Mutation of the Xylanase regulator 1 causes a glucose blind hydrolase expressing phenotype in industrially used Trichoderma strains. Biotechnol Biofuels 6:62

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Li J, Wang J, Wang S, Xing M, Yu S, Liu G (2012) Achieving efficient protein expression in Trichoderma reesei by using strong constitutive promoters. Microb Cell Fact 11:84–94

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Li Y, Liu C, Bai F, Zhao X (2016) Overproduction of cellulase by Trichoderma reesei RUT C30 through batch-feeding of synthesized low-cost sugar mixture. Bioresour Technol 216:503–510

    Article  PubMed  CAS  Google Scholar 

  • Mandels M, Weber J (1969) The production of cellulases. Adv Chem Ser 95:391–414

    Article  CAS  Google Scholar 

  • Peterson R, Nevalainen H (2012) Trichoderma reesei RUT-C30–thirty years of strain improvement. Microbiology 158:58–68

    Article  PubMed  CAS  Google Scholar 

  • Sethia PP, Rao KK, Noronha SB (2014) A dps promoter based expression system for improved solubility of expressed proteins in Escherichia coli. Biotechnol Bioprocess 19:790–797

    Article  CAS  Google Scholar 

  • Stricker AR, Trefflinger P, Aro N, Penttilä M, Mach RL (2008) Role of Ace2 (Activator of Cellulases 2) within the xyn2 transcriptosome of Hypocrea jecorina. Fungal Genet Biol 45:436–445

    Article  PubMed  CAS  Google Scholar 

  • Takashima S, Iikura H, Nakamura A, Hidaka M, Masaki H, Uozumi T (1998) Overproduction of recombinant Trichoderma reesei cellulases by Aspergillus oryzae and their enzymatic properties. J Biotechnol 65:163–171

    Article  PubMed  CAS  Google Scholar 

  • Wang W, Meng F, Liu P, Yang S, Wei D (2014) Construction of a promoter collection for genes co-expression in filamentous fungus Trichoderma reesei. J Ind Microbiol Biotechnol 41:1709–1718

    Article  PubMed  CAS  Google Scholar 

  • Woodward J, Carmichael JS, Capps KM, Herrmann PC, Lee NE (1990) The competitive inhibition of Trichoderma reesei C30 cellobiohydrolase I by guanidine hydrochloride. FEBS Lett 270:143

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Li X, Xia L (2015) Heterologous expression of an alkali and thermotolerant lipase from Talaromyces thermophilus in Trichoderma reesei. Appl Biochem Biotech 176:1722–1735

    Article  CAS  Google Scholar 

  • Zhang L, Zhang S, Jiang X, Wei W, Wang W, Wei D (2016a) A novel host-vector system for heterologous protein co-expression and purification in the Trichoderma reesei industrial strain RUT-C30. Biotechnol Lett 38:89–96

    Article  PubMed  CAS  Google Scholar 

  • Zhang L, Zhao X, Zhang G, Zhang J, Wang X, Zhang S, Wang W, Wei D (2016b) Light-inducible genetic engineering and control of non-homologous end-joining in industrial eukaryotic microorganisms: LML 3.0 and OFN 1.0. Sci Rep 6:20761

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhang X, Li Y, Zhao X, Bai F (2017) Constitutive cellulase production from glucose using the recombinant Trichoderma reesei strain overexpressing an artificial transcription activator. Bioresour Technol 223:317–322

    Article  PubMed  CAS  Google Scholar 

  • Zou G, Shi S, Jiang Y, Brink J, Vries R, Chen L, Zhang J, Ma L, Wang C, Zhou Z (2012) Construction of a cellulase hyper-expression system in Trichoderma reesei by promoter and enzyme engineering. Microb Cell Fact 11:21

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

Acknowledgements

This research was supported by the National Natural Science Foundation of China [No. C010302-31500066] and the Fundamental Research Funds for the Central Universities [No. 222201714053].

Supporting information

Supplementary Table 1—Primers for pXMG, pX1rfp, pX2rfp, pX1egv3 and pX2egv3 constructed in this study.

Supplementary Figure 1—Schematic for identification of gene size and single-copy DNA integration in the genome of the transformants.

Supplementary Figure 2—SDS-PAGE analysis of extracellular proteins secreted by T. reesei RUT C30 and TXMG in different cultivation strategies.

Author information

Authors and Affiliations

  1. New World Institute of Biotechnology, State Key Lab of Bioreactor Engineering, East China University of Science and Technology, P.O.B. 311, 130 Meilong Road, Shanghai, 200237, China

    Jiajia Zhang, Chuan Wu, Wei Wang & Dongzhi Wei

  2. State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China

    Wei Wang

Authors
  1. Jiajia Zhang
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  2. Chuan Wu
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  3. Wei Wang
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  4. Wei Wang
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  5. Dongzhi Wei
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Corresponding author

Correspondence to Wei Wang.

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The author(s) declare that they have no competing interests.

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Zhang, J., Wu, C., Wang, W. et al. A versatile Trichoderma reesei expression system for the production of heterologous proteins. Biotechnol Lett 40, 965–972 (2018). https://doi.org/10.1007/s10529-018-2548-x

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  • DOI: https://doi.org/10.1007/s10529-018-2548-x

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