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Cell surface display of a β-glucosidase employing the type V secretion system on ethanologenic Escherichia coli for the fermentation of cellobiose to ethanol

  • Iván Muñoz-Gutiérrez
  • Ricardo Oropeza
  • Guillermo Gosset
  • Alfredo Martinez
Bioenergy/Biofuels/Biochemicals

Abstract

We used the autodisplay system AIDA-I, which belongs to the type V secretion system (TVSS), to display the β-glucosidase BglC from Thermobifida fusca on the outer membrane of the ethanologenic Escherichia coli strain MS04 (MG1655 ∆pflB, ∆adhE, ∆frdA, ∆xylFGH, ∆ldhA, PpflB::pdc Zm -adhB Zm ). MS04 that was transformed with the plasmid pAIDABglCRHis showed cellobiase activity (171 U/gCDW) and fermented 40 g/l cellobiose in mineral medium in 60 h with an ethanol yield of 81 % of the theoretical maximum. Whole-cell protease treatment, SDS-PAGE, and Western-blot analysis demonstrated that BglC was attached to the external surface of the outer membrane of MS04. When attached to the cells, BglC showed 93.3 % relative activity in the presence of 40 g/l ethanol and retained 100 % of its activity following 2 days of incubation at 37 °C with the same ethanol concentration. This study shows the potential of the TVSS (AIDA-I) and BglC as tools for the production of lignocellulosic bio-commodities.

Keywords

Cellobiose Bioethanol Escherichia coli Type V secretion system AIDA-I Thermobifida fusca β-glucosidase 

Notes

Acknowledgments

We thank Dr. Thomas F. Meyer from Max Planck Institute (Infection Biology) for providing pJM7 plasmid, Dr. David B. Wilson (Department of Molecular Biology and Genetics, Cornell University) for providing pNS6 plasmid, and Luz María Martínez, Mercedes Enzaldo, Georgina Hernández, Omar Arriaga and Shirley Ainsworth for technical support. This work was supported by the Mexican Council of Science and Technology (CONACyT) technological innovation grants: PETRAMIN 2010-13879, 2011-154298, and 2012-184417; and from the Universidad Nacional Autónoma de México: grant DGAPA/PAPIIT/UNAM IT200312-2.

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2012

Authors and Affiliations

  • Iván Muñoz-Gutiérrez
    • 1
  • Ricardo Oropeza
    • 2
  • Guillermo Gosset
    • 1
  • Alfredo Martinez
    • 1
  1. 1.Departamento de Ingeniería Celular y Biocatálisis, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMexico
  2. 2.Departamento de Microbiología Molecular, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMexico

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