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Optimization of the culture medium composition using response surface methodology for new recombinant cyprosin B production in bioreactor for cheese production

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Abstract

The optimization of culture medium composition was carried out for improvement the recombinant cyprosin B production, an enzyme with high milk-clotting activity. Response surface methodology (RSM) was applied to evaluate the effect of variables namely glucose, yeast extract (YE) and bactopeptone present in the culture medium, used for recombinant cyprosin B production by transformed Saccharomyces cerevisiae BJ1991 strain in shake-flask and bioreactor culture conditions. The central composite experimental design (CCD) was adopted to derive a statistical model for optimizing the composition of the fermentation medium. The optimal concentration estimated for each variable related to a theoretical maximum of cyprosin B activity (488 U mL−1) was 30 g L−1 glucose, 15 g L−1 YE and 27 g L−1 bactopeptone. The optimized medium composition, based on empirical model, led to a cyprosin B activity of 519 U mL−1, which corresponds to an increase of 46%. The fermentation using optimized culture medium in a 5-L bioreactor allowed a significant increase in biomass (82%) and recombinant cyprosin B production (139%). The improvement in the recombinant cyprosin B production after optimization process can be considered adequate for large-scale applications, and the clotting activity of cyprosin B account for their use in industrial cheese making.

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Acknowledgments

We thank Dr. Filomena Calixto for providing S. cerevisiae BJ1991 strain transformed with CYPRO11 gene. PN Sampaio was supported by PRAXIS XXI Ph.D Fellowship (SFRH/BD/8780/2002) from Foundation for Science and Technology, Portugal. L Sousa was supported by the research projects PTDC/MAT/64353/2006 and FCT/OE.

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Authors and Affiliations

  1. BioFIG, Unit of Molecular Biology & Plant Biotechnology, Plant Systems Biology Laboratory, Institute of Applied Science and Technology, Faculty of Sciences of University of Lisbon, Campo Grande, 1749-016, Lisbon, Portugal

    Pedro N. Sampaio & Maria Salomé Pais

  2. Institute of Biotechnology and Bioengineering, Chemical and Biological Engineering Center, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001, Lisbon, Portugal

    Pedro N. Sampaio & Luís Pina Fonseca

  3. Faculty of Engineering, Catholic University of Portugal, Estrada Octávio Pato, 2635-631, Rio de Mouro, Portugal

    Cecília R. C. Calado

  4. Department of Statistics and Operations Research, Center of Statistics and Applications (CEAUL), Faculty of Sciences of University of Lisbon, 1749-016, Lisbon, Portugal

    Lisete Sousa

  5. Agricultural, Food and Nutritional Science Department, Faculty of Agricultural, Life & Environmental Sciences, University of Alberta, 410 Agriculture Forestry Bldg, Edmonton, AB, T6G 2P5, Canada

    David C. Bressler

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  1. Pedro N. Sampaio
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  2. Cecília R. C. Calado
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  3. Lisete Sousa
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  4. David C. Bressler
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  5. Maria Salomé Pais
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  6. Luís Pina Fonseca
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Correspondence to Pedro N. Sampaio.

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Sampaio, P.N., Calado, C.R.C., Sousa, L. et al. Optimization of the culture medium composition using response surface methodology for new recombinant cyprosin B production in bioreactor for cheese production. Eur Food Res Technol 231, 339–346 (2010). https://doi.org/10.1007/s00217-010-1281-z

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  • DOI: https://doi.org/10.1007/s00217-010-1281-z

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