Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR
The quality of functional food products designed for the prevention of degenerative diseases can be affected by the incorporation of bioactive compounds. In many types of cheese, the performance of starter microorganisms is critical for optimal elaboration and for providing potential probiotic benef...
| Autores principales: | , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.12123/1743 https://www.sciencedirect.com/science/article/pii/S0168160516302951?via%3Dihub https://doi.org/10.1016/j.ijfoodmicro.2016.06.003 |
| _version_ | 1855482935961976832 |
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| author | Pega, Juan Franco Rizzo, Sergio Anibal Perez, Carolina Daiana Rossetti, Luciana Diaz, Gabriela Ruzal, S.M. Nanni, Mariana Descalzo, Adriana Maria |
| author_browse | Descalzo, Adriana Maria Diaz, Gabriela Nanni, Mariana Pega, Juan Franco Perez, Carolina Daiana Rizzo, Sergio Anibal Rossetti, Luciana Ruzal, S.M. |
| author_facet | Pega, Juan Franco Rizzo, Sergio Anibal Perez, Carolina Daiana Rossetti, Luciana Diaz, Gabriela Ruzal, S.M. Nanni, Mariana Descalzo, Adriana Maria |
| author_sort | Pega, Juan Franco |
| collection | INTA Digital |
| description | The quality of functional food products designed for the prevention of degenerative diseases can be affected by the incorporation of bioactive compounds. In many types of cheese, the performance of starter microorganisms is critical for optimal elaboration and for providing potential probiotic benefits. Phytosterols are plant lipophilic triterpenes that have been used for the design of functional dairy products because of their ability to lower serum cholesterol levels in humans. However, their effect on the starter culture behavior during cheesemaking has not yet been studied. Here, we followed DNA and RNA kinetics of the bacterium Streptococcus thermophilus, an extensively used dairy starter with probiotic potential, during industrial production of a functional, semi-soft, reduced-fat cheese containing phytosterol esters and alpha-tocopherol as bioactive compounds. For this purpose, real-time quantitative PCR (qPCR) and reverse transcription-qPCR (RT-qPCR) assays were optimized and applied to samples obtained during the manufacture and ripening of functional and control cheeses. An experimental set-up was used to evaluate the detection threshold of free nucleic acids for extraction protocols based on pelleted microorganisms. To our knowledge, this straight-forward approach provides the first experimental evidence indicating that DNA is not a reliable marker of cell integrity, whereas RNA may constitute a more accurate molecular signature to estimate both bacterial viability and metabolic activity. Compositional analysis revealed that the bioactive molecules were effectively incorporated into the cheese matrix, at levels considered optimal to exert their biological action. The starter S. thermophilus was detected by qPCR and RT-qPCR during cheese production at the industrial level, from at least 30 min after its inoculation until 81 days of ripening, supporting the possible role of this species in shaping organoleptic profiles. We also showed for the first time that the addition of phytosterols at functional concentrations, not only did not affect starter performance but also correlated with a significant increase in target DNA and cDNA levels in most of the time points evaluated throughout cheesemaking. Therefore, these findings suggest that the growth and metabolism of S. thermophilus may be enhanced by the incorporation of these biologically active molecules during cheese production, providing important information for the industrial design of novel fermented foods. |
| format | Artículo |
| id | INTA1743 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA17432018-07-05T18:04:30Z Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR Pega, Juan Franco Rizzo, Sergio Anibal Perez, Carolina Daiana Rossetti, Luciana Diaz, Gabriela Ruzal, S.M. Nanni, Mariana Descalzo, Adriana Maria Streptococcus Thermophilus PCR Alimentos Sanos Queso Health Foods Cheese Phytosterols Tocopherols Fitosteroles Tocoferoles Alimentos Funcionales The quality of functional food products designed for the prevention of degenerative diseases can be affected by the incorporation of bioactive compounds. In many types of cheese, the performance of starter microorganisms is critical for optimal elaboration and for providing potential probiotic benefits. Phytosterols are plant lipophilic triterpenes that have been used for the design of functional dairy products because of their ability to lower serum cholesterol levels in humans. However, their effect on the starter culture behavior during cheesemaking has not yet been studied. Here, we followed DNA and RNA kinetics of the bacterium Streptococcus thermophilus, an extensively used dairy starter with probiotic potential, during industrial production of a functional, semi-soft, reduced-fat cheese containing phytosterol esters and alpha-tocopherol as bioactive compounds. For this purpose, real-time quantitative PCR (qPCR) and reverse transcription-qPCR (RT-qPCR) assays were optimized and applied to samples obtained during the manufacture and ripening of functional and control cheeses. An experimental set-up was used to evaluate the detection threshold of free nucleic acids for extraction protocols based on pelleted microorganisms. To our knowledge, this straight-forward approach provides the first experimental evidence indicating that DNA is not a reliable marker of cell integrity, whereas RNA may constitute a more accurate molecular signature to estimate both bacterial viability and metabolic activity. Compositional analysis revealed that the bioactive molecules were effectively incorporated into the cheese matrix, at levels considered optimal to exert their biological action. The starter S. thermophilus was detected by qPCR and RT-qPCR during cheese production at the industrial level, from at least 30 min after its inoculation until 81 days of ripening, supporting the possible role of this species in shaping organoleptic profiles. We also showed for the first time that the addition of phytosterols at functional concentrations, not only did not affect starter performance but also correlated with a significant increase in target DNA and cDNA levels in most of the time points evaluated throughout cheesemaking. Therefore, these findings suggest that the growth and metabolism of S. thermophilus may be enhanced by the incorporation of these biologically active molecules during cheese production, providing important information for the industrial design of novel fermented foods. Fil: Pega, Juan Franco. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rizzo, Sergio Anibal. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina Fil: Perez, Carolina Daiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rossetti, Luciana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina Fil: Diaz, Gabriela Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina Fil: Ruzal, Sandra Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales; Argentina Fil: Nanni, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina Fil: Descalzo, Adriana Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA).LABINTEX-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD). Département PERSYST; Francia 2017-11-10T13:02:10Z 2017-11-10T13:02:10Z 2016-09 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion http://hdl.handle.net/20.500.12123/1743 https://www.sciencedirect.com/science/article/pii/S0168160516302951?via%3Dihub 0168-1605 (Print) 1879-3460 (Online) https://doi.org/10.1016/j.ijfoodmicro.2016.06.003 eng info:eu-repo/semantics/restrictedAccess application/pdf International journal of food microbiology 232 : 117-125. (September 2016) |
| spellingShingle | Streptococcus Thermophilus PCR Alimentos Sanos Queso Health Foods Cheese Phytosterols Tocopherols Fitosteroles Tocoferoles Alimentos Funcionales Pega, Juan Franco Rizzo, Sergio Anibal Perez, Carolina Daiana Rossetti, Luciana Diaz, Gabriela Ruzal, S.M. Nanni, Mariana Descalzo, Adriana Maria Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR |
| title | Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR |
| title_full | Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR |
| title_fullStr | Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR |
| title_full_unstemmed | Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR |
| title_short | Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR |
| title_sort | effect of the addition of phytosterols and tocopherols on streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qpcr and rt qpcr |
| topic | Streptococcus Thermophilus PCR Alimentos Sanos Queso Health Foods Cheese Phytosterols Tocopherols Fitosteroles Tocoferoles Alimentos Funcionales |
| url | http://hdl.handle.net/20.500.12123/1743 https://www.sciencedirect.com/science/article/pii/S0168160516302951?via%3Dihub https://doi.org/10.1016/j.ijfoodmicro.2016.06.003 |
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