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Heterologous expression of Fusarium oxysporum tomatinase in Saccharomyces cerevisiae increases its resistance to saponins and improves ethanol production during the fermentation of Agave tequilana Weber var. azul and Agave salmiana must

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Abstract

This paper describes the effect of the heterologous expression of tomatinase from Fusarium oxysporum f. sp lycopersici in Saccharomyces cerevisiae. The gene FoTom1 under the control of the S. cerevisiae phosphoglycerate kinase (PGK1) promoter was cloned into pYES2. S. cerevisiae strain Y45 was transformed with this vector and URA3 transformant strains were selected for resistance to α-tomatine. Two transformants were randomly selected for further study (designated Y45-1 and Y45-2). Control strain Y45 was inhibited at 50 μM α-tomatine, in contrast, transformants Y45-1 and Y45-2 did not show inhibition at 200 μM. Tomatinase activity was detected by HPLC monitoring tomatine disappearance and tomatidine appearance in the supernatants of culture medium. Maximum tomatinase activity was observed in the transformants after 6 h, remaining constant during the following 24 h. No tomatinase activity was detected in the parental strain. Moreover, the transformants were able to grow and produce ethanol in a mix of Agave tequilana Weber var. azul and Agave salmiana must, contrary to the Y45 strain which was unable to grow and ferment under these conditions.

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Acknowledgements

This research was supported by the Consejo Nacional de la Ciencia y Tecnología de México (CONACYT, Project 41148).

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Authors and Affiliations

  1. Centro de Investigación y Asistenta Tecnológica del Estado de Jalisco (CIATEJ), Unidad de Biotecnología, Av. Normalistas 800, Col. Colinas de la Normal, C.P. 44270, Guadalajara, Jalisco, Mexico

    Luis Alberto Cira, Melesio Gutiérrez & Jesús Ramírez

  2. Instituto de Investigación en Biología Experimental, Universidad de Guanajuato, Guanajuato, Mexico

    Gloria Angélica González & Juan Carlos Torres

  3. Centro Universitario de Ciencias exactas e Ingeniería de la Universidad de Guadalajara, Av. Marcelino García Barragán y Calzada Olímpica S.R., C.P. 44420, Guadalajara, Jalisco, Mexico

    Carlos Pelayo

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  1. Luis Alberto Cira
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  2. Gloria Angélica González
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  3. Juan Carlos Torres
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  4. Carlos Pelayo
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Correspondence to Jesús Ramírez.

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Cira, L.A., González, G.A., Torres, J.C. et al. Heterologous expression of Fusarium oxysporum tomatinase in Saccharomyces cerevisiae increases its resistance to saponins and improves ethanol production during the fermentation of Agave tequilana Weber var. azul and Agave salmiana must. Antonie van Leeuwenhoek 93, 259–266 (2008). https://doi.org/10.1007/s10482-007-9200-4

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