Archives of Microbiology

, Volume 199, Issue 4, pp 605–611 | Cite as

Expression of a codon-optimized β-glucosidase from Cellulomonas flavigena PR-22 in Saccharomyces cerevisiae for bioethanol production from cellobiose

  • Francisco Javier Ríos-Fránquez
  • Enrique González-Bautista
  • Teresa Ponce-Noyola
  • Ana Carmela Ramos-Valdivia
  • Héctor Mario Poggi-Varaldo
  • Jaime García-Mena
  • Alfredo Martinez
Original Paper

Abstract

Bioethanol is one of the main biofuels produced from the fermentation of saccharified agricultural waste; however, this technology needs to be optimized for profitability. Because the commonly used ethanologenic yeast strains are unable to assimilate cellobiose, several efforts have been made to express cellulose hydrolytic enzymes in these yeasts to produce ethanol from lignocellulose. The C. flavigenabglA gene encoding β-glucosidase catalytic subunit was optimized for preferential codon usage in S. cerevisiae. The optimized gene, cloned into the episomal vector pRGP-1, was expressed, which led to the secretion of an active β-glucosidase in transformants of the S. cerevisiae diploid strain 2-24D. The volumetric and specific extracellular enzymatic activities using pNPG as substrate were 155 IU L−1 and 222 IU g−1, respectively, as detected in the supernatant of the cultures of the S. cerevisiae RP2-BGL transformant strain growing in cellobiose (20 g L−1) as the sole carbon source for 48 h. Ethanol production was 5 g L−1 after 96 h of culture, which represented a yield of 0.41 g g−1 of substrate consumed (12 g L−1), equivalent to 76% of the theoretical yield. The S. cerevisiae RP2-BGL strain expressed the β-glucosidase extracellularly and produced ethanol from cellobiose, which makes this microorganism suitable for application in ethanol production processes with saccharified lignocellulose.

Keywords

Saccharomyces cerevisiae Cellulomonas flavigena β-Glucosidase expression Cellobiose Bioethanol 

Notes

Acknowledgements

This work was supported by the Mexican Council of Science and Technology (CONACyT), Grant CB-2014/236895, and the Bioenergy Thematic Network (Red Temática de Bioenergía), Grant 260457. The authors thank Odilia Pérez-Avalos and Gustavo Medina-Mendoza for technical assistance. FJ Ríos-Fránquez received a Ph. D scholarship from CONACyT-México. The authors have no conflict of interest to declare.

Supplementary material

203_2016_1333_MOESM1_ESM.tiff (349 kb)
Supplementary material 1 (TIFF 349 KB)
203_2016_1333_MOESM2_ESM.docx (22 kb)
Supplementary material 2 (DOCX 22 KB)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Francisco Javier Ríos-Fránquez
    • 1
  • Enrique González-Bautista
    • 1
  • Teresa Ponce-Noyola
    • 1
  • Ana Carmela Ramos-Valdivia
    • 1
  • Héctor Mario Poggi-Varaldo
    • 1
  • Jaime García-Mena
    • 2
  • Alfredo Martinez
    • 3
  1. 1.Departamento de Biotecnología y BioingenieríaCinvestav-IPNMexico CityMexico
  2. 2.Departamento de Genética y Biología MolecularCinvestav-IPNMexico CityMexico
  3. 3.Departamento de Ingeniería Celular y Biocatálisis, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMexico

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