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Genetic characterization and construction of an auxotrophic strain of Saccharomyces cerevisiae JP1, a Brazilian industrial yeast strain for bioethanol production

  • Genetics and Molecular Biology of Industrial Organisms
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Used for millennia to produce beverages and food, Saccharomyces cerevisiae also became a workhorse in the production of biofuels, most notably bioethanol. Yeast strains have acquired distinct characteristics that are the result of evolutionary adaptation to the stresses of industrial ethanol production. JP1 is a dominant industrial S. cerevisiae strain isolated from a sugarcane mill and is becoming increasingly popular for bioethanol production in Brazil. In this work, we carried out the genetic characterization of this strain and developed a set of tools to permit its genetic manipulation. Using flow cytometry, mating type, and sporulation analysis, we verified that JP1 is diploid and homothallic. Vectors with dominant selective markers for G418, hygromycin B, zeocin, and ρ-fluoro-dl-phenylalanine were used to successfully transform JP1 cells. Also, an auxotrophic ura3 mutant strain of JP1 was created by gene disruption using integration cassettes with dominant markers flanked by loxP sites. Marker excision was accomplished by the Cre/loxP system. The resulting auxotrophic strain was successfully transformed with an episomal vector that allowed green fluorescent protein expression.

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Acknowledgments

The research was supported by Petrobras, CNPq and Capes (Brazil). The authors thank Dr. Olssen, Dr. Falcon, and Dr. González for donation of plasmid vectors. We are in indebted to Dr. Marcos Morais (Universidade Federal de Pernambuco) for critical review of this manuscript.

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Authors and Affiliations

  1. Centro de Biotecnologia Molecular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70910-900, Brazil

    Viviane Castelo Branco Reis, Osmar de Souza Oliveira Neto, Vinícius Daniel Ferreira Batista, Lidia Maria Pepe de Moraes & Fernando Araripe Gonçalves Torres

  2. Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Brasília, DF, 709790-160, Brazil

    André Moraes Nicola

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  1. Viviane Castelo Branco Reis
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  2. André Moraes Nicola
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  3. Osmar de Souza Oliveira Neto
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Correspondence to Fernando Araripe Gonçalves Torres.

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Reis, V.C.B., Nicola, A.M., de Souza Oliveira Neto, O. et al. Genetic characterization and construction of an auxotrophic strain of Saccharomyces cerevisiae JP1, a Brazilian industrial yeast strain for bioethanol production. J Ind Microbiol Biotechnol 39, 1673–1683 (2012). https://doi.org/10.1007/s10295-012-1170-5

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