Molecular analysis of the interactions between starter cultures during wine fermentation
In recent years, the wine industry has been facing major challenges such as climate change, one of the consequences of which is the increase in the concentration of sugars in grapes at harvest time and, consequently, in the alcohol content of wines. On the other hand, the widespread use of Saccharom...
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Universidad de La Rioja (España)
2021
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In recent years, the wine industry has been facing major challenges such as climate change, one of the consequences of which is the increase in the concentration of sugars in grapes at harvest time and, consequently, in the alcohol content of wines. On the other hand, the widespread use of Saccharomyces cerevisiae strains as fermentation starters has led to a perception of uniformity or standardization of the aromatic profile of wines. In both cases, part of the proposals for alcohol reduction and product diversification involve the use of starter cultures of alternative species to S. cerevisiae, commonly referred to as non-Saccharomyces. These new starter cultures are usually combined with some strain of S. cerevisiae in order to avoid stuck fermentation. The use of more than one species as starter culture implies that there is more than one population with a similar number of individuals performing the fermentation process simultaneously. In this way, the interactions that are established between them acquire a relevance that until now had not been valued. Today, it is widely accepted that the outcome of a fermentation process involving two or more inoculated yeast species will be different from the weighted average of the corresponding individual cultures. Interspecific interactions between wine yeasts take place at various levels, including interference competition, exploitation competition, and exchange of metabolic intermediates, among others. Some interactions could be a simple consequence of the metabolism of each yeast in a context where metabolic intermediates and end products of other yeasts are present. However, there are clear indications, in some cases, of specific recognition between interacting yeasts. In general terms, this doctoral thesis aims to deepen the knowledge of how microbial communities behave in wine, analyzing such behavior mainly from a transcriptomic and proteomic point of view, trying to anticipate their biological responses in this complex matrix. In this sense, the first chapter of the thesis focuses on the practical impact of using oenological yeasts that present the prion [GAR+] element in the fermentation of natural grape must. Parameters such as real alcohol reduction, volatile acidity produced, fermentation kinetics and fermentation time under both aerobic and anaerobic conditions are evaluated. In addition, the isolation frequency, heritability and penetrance values of this type of prion in six industrial strains of S. cerevisiae with different genetic backgrounds are analyzed. Among the main results, the low phenotypic penetrance of this epigenetic element and the absence of reduction in alcoholic yield are highlighted. The second chapter of the thesis focuses on the study of short-term yeast-yeast interactions to prevent changes in nutrient composition from affecting yeast metabolic activity. The transcriptional response of S. cerevisiae to co-culture, in only three hours, with the yeast Metschnikowia pulcherrima has been evaluated. In addition, the responses obtained in this chapter have been compared with those obtained in a previous work carried out by this research group in which the short-term response of S. cerevisiae was also analyzed, but against co-culture with yeasts of the species Torulaspora delbrueckii, Candida sake and Hanseniaspora uvarum. The results show that S. cerevisiae responds to M. pulcherrima in a more intense manner than to other yeast species, involving a repression of respiratory activity, in addition to the induction of glycolysis, which is common to all the responses analyzed. In turn, the third and fourth chapters of this PhD thesis focus on the study of EVs as complex structures with potential participation in the communication processes between yeast cells under oenological fermentation conditions. Specifically, the third chapter not only evaluates whether oenological yeasts are capable of producing EVs in an oenological context, but also aims to analyze which proteins are mostly present in EV-enriched fractions by means of a proteomic analysis. This study has shown, as described in the literature, an enrichment in proteins related to the cell wall and glycolysis. In addition to the hypothesis that EVs act as signal carriers in the interactions between different wine yeast species, the fourth chapter of this doctoral thesis evaluates whether the protein content of EVs is, in turn, affected by these interactions. The results show a clear impact of co-cultures on protein composition in all cases. In addition, although no firm conclusions can be drawn, there are indications that some yeasts shown common factors in their response to different competitors, while others induce in other yeasts a response that shows some commmon factor among several of them. Taken together, the results shown in these last two chapters point to a potential role of EVs in communication between different oenological yeast species. In general terms, this doctoral thesis contributes not only to the fundamental knowledge of long-term ecological interactions, but also provides a glimpse of their potential industrial applicability. In this sense, a precise knowledge of the interaction mechanisms would allow the design of new mixed starter cultures based on Saccharomyces and non-Saccharomyces yeasts, as well as, the optimal conditions for their application. |
| author2 |
González García, Ramón (null) |
| author_facet |
González García, Ramón (null) Mencher Beltran, Ana |
| format |
text (thesis) |
| author |
Mencher Beltran, Ana |
| spellingShingle |
Mencher Beltran, Ana Molecular analysis of the interactions between starter cultures during wine fermentation |
| author_sort |
Mencher Beltran, Ana |
| title |
Molecular analysis of the interactions between starter cultures during wine fermentation |
| title_short |
Molecular analysis of the interactions between starter cultures during wine fermentation |
| title_full |
Molecular analysis of the interactions between starter cultures during wine fermentation |
| title_fullStr |
Molecular analysis of the interactions between starter cultures during wine fermentation |
| title_full_unstemmed |
Molecular analysis of the interactions between starter cultures during wine fermentation |
| title_sort |
molecular analysis of the interactions between starter cultures during wine fermentation |
| publisher |
Universidad de La Rioja (España) |
| publishDate |
2021 |
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https://dialnet.unirioja.es/servlet/oaites?codigo=296473 |
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AT mencherbeltranana molecularanalysisoftheinteractionsbetweenstarterculturesduringwinefermentation |
| _version_ |
1718346698840866816 |
| spelling |
oai-TES00000230142021-11-20Molecular analysis of the interactions between starter cultures during wine fermentationMencher Beltran, AnaIn recent years, the wine industry has been facing major challenges such as climate change, one of the consequences of which is the increase in the concentration of sugars in grapes at harvest time and, consequently, in the alcohol content of wines. On the other hand, the widespread use of Saccharomyces cerevisiae strains as fermentation starters has led to a perception of uniformity or standardization of the aromatic profile of wines. In both cases, part of the proposals for alcohol reduction and product diversification involve the use of starter cultures of alternative species to S. cerevisiae, commonly referred to as non-Saccharomyces. These new starter cultures are usually combined with some strain of S. cerevisiae in order to avoid stuck fermentation. The use of more than one species as starter culture implies that there is more than one population with a similar number of individuals performing the fermentation process simultaneously. In this way, the interactions that are established between them acquire a relevance that until now had not been valued. Today, it is widely accepted that the outcome of a fermentation process involving two or more inoculated yeast species will be different from the weighted average of the corresponding individual cultures. Interspecific interactions between wine yeasts take place at various levels, including interference competition, exploitation competition, and exchange of metabolic intermediates, among others. Some interactions could be a simple consequence of the metabolism of each yeast in a context where metabolic intermediates and end products of other yeasts are present. However, there are clear indications, in some cases, of specific recognition between interacting yeasts. In general terms, this doctoral thesis aims to deepen the knowledge of how microbial communities behave in wine, analyzing such behavior mainly from a transcriptomic and proteomic point of view, trying to anticipate their biological responses in this complex matrix. In this sense, the first chapter of the thesis focuses on the practical impact of using oenological yeasts that present the prion [GAR+] element in the fermentation of natural grape must. Parameters such as real alcohol reduction, volatile acidity produced, fermentation kinetics and fermentation time under both aerobic and anaerobic conditions are evaluated. In addition, the isolation frequency, heritability and penetrance values of this type of prion in six industrial strains of S. cerevisiae with different genetic backgrounds are analyzed. Among the main results, the low phenotypic penetrance of this epigenetic element and the absence of reduction in alcoholic yield are highlighted. The second chapter of the thesis focuses on the study of short-term yeast-yeast interactions to prevent changes in nutrient composition from affecting yeast metabolic activity. The transcriptional response of S. cerevisiae to co-culture, in only three hours, with the yeast Metschnikowia pulcherrima has been evaluated. In addition, the responses obtained in this chapter have been compared with those obtained in a previous work carried out by this research group in which the short-term response of S. cerevisiae was also analyzed, but against co-culture with yeasts of the species Torulaspora delbrueckii, Candida sake and Hanseniaspora uvarum. The results show that S. cerevisiae responds to M. pulcherrima in a more intense manner than to other yeast species, involving a repression of respiratory activity, in addition to the induction of glycolysis, which is common to all the responses analyzed. In turn, the third and fourth chapters of this PhD thesis focus on the study of EVs as complex structures with potential participation in the communication processes between yeast cells under oenological fermentation conditions. Specifically, the third chapter not only evaluates whether oenological yeasts are capable of producing EVs in an oenological context, but also aims to analyze which proteins are mostly present in EV-enriched fractions by means of a proteomic analysis. This study has shown, as described in the literature, an enrichment in proteins related to the cell wall and glycolysis. In addition to the hypothesis that EVs act as signal carriers in the interactions between different wine yeast species, the fourth chapter of this doctoral thesis evaluates whether the protein content of EVs is, in turn, affected by these interactions. The results show a clear impact of co-cultures on protein composition in all cases. In addition, although no firm conclusions can be drawn, there are indications that some yeasts shown common factors in their response to different competitors, while others induce in other yeasts a response that shows some commmon factor among several of them. Taken together, the results shown in these last two chapters point to a potential role of EVs in communication between different oenological yeast species. In general terms, this doctoral thesis contributes not only to the fundamental knowledge of long-term ecological interactions, but also provides a glimpse of their potential industrial applicability. In this sense, a precise knowledge of the interaction mechanisms would allow the design of new mixed starter cultures based on Saccharomyces and non-Saccharomyces yeasts, as well as, the optimal conditions for their application.Durante los últimos años, la industria del vino se está enfrentando a retos importantes como el cambio climático, una de cuyas consecuencias es el aumento de la concentración de azúcares en la uva en el momento de la vendimia y, por consiguiente, del grado alcohólico en los vinos. Por otra parte, el uso generalizado de cepas de Saccharomyces cerevisiae como iniciadores de la fermentación ha llevado a una percepción de uniformidad o estandarización del perfil aromático de los vinos. En ambos casos, parte de las propuestas para la reducción del grado alcohólico y la diversificación de productos pasan por el uso de cultivos iniciadores de especies alternativas a S. cerevisiae, comúnmente denominados como no-Saccharomyces. Estos nuevos cultivos iniciadores se combinan habitualmente con alguna cepa de S. cerevisiae, con el fin de evitar paradas de fermentación. El uso de más de una especie como cultivo iniciador, supone que haya más de una población con un número similar de individuos que realizan el proceso de fermentación simultáneamente. De esta manera, las interacciones que se establecen entre ellas adquieren una relevancia que hasta ahora no había sido valorada. Hoy en día, está ampliamente aceptado que el resultado de un proceso de fermentación en el que participan dos o más especies de levaduras inoculadas será diferente de la media ponderada de los correspondientes cultivos individuales. Las interacciones interespecíficas entre las levaduras del vino tienen lugar a varios niveles, incluyendo la competencia por interferencia, la competencia por explotación y el intercambio de intermediarios metabólicos, entre otros. Algunas interacciones podrían ser una simple consecuencia del metabolismo de cada levadura en un contexto en el que están presentes los intermediarios metabólicos y los productos finales de otras levaduras. Sin embargo, hay indicios claros, en algunos casos, de un reconocimiento específico entre las levaduras que interactúan. De forma general, esta tesis doctoral pretende profundizar en el conocimiento de cómo se comportan las comunidades microbianas en el vino, analizando dicho comportamiento principalmente desde un punto de vista transcriptómico y proteómico, tratando de anticipar sus respuestas biológicas en esta compleja matriz. En este sentido, el primer capítulo de la tesis se centra en el impacto práctico que conlleva el utilizar levaduras enológicas que presentan el elemento prion [GAR+] en la fermentación del mosto natural de uva. Se evalúan parámetros como la reducción real de alcohol, la acidez volátil producida, la cinética y tiempo de fermentación, tanto en condiciones aerobias como anaerobias. Además, se analiza la frecuencia de aislamiento, la heredabilidad y valores de penetrancia de este tipo de prion en seis cepas industriales de S. cerevisiae con diferentes fondos genéticos. Entre los resultados principales se destaca la baja penetrancia fenotípica de este elemento epigenético, y la ausencia de reducción del rendimiento alcohólico. El segundo capítulo de la tesis se centra en el estudio de las interacciones levadura-levadura a corto plazo para evitar que los cambios en la composición de nutrientes afecten a la actividad metabólica de la levadura. Se ha evaluado la respuesta transcripcional de S. cerevisiae al co-cultivo, en tan solo tres horas, con la levadura Metschnikowia pulcherrima. Además, se han comparado las respuestas obtenidas en este capítulo con las obtenidas en un trabajo anterior realizado por este grupo de investigación en el que se analizaba la respuesta a corto plazo también de S. cerevisiae, pero frente al co-cultivo con levaduras de las especies Torulaspora delbrueckii, Candida sake y Hanseniaspora uvarum. Los resultados muestran que S. cerevisiae responde a M. pulcherrima de una manera más intensa que a otras especies de levaduras, implicando una represión de la actividad respiratoria, además de la inducción de la glicolisis, que es común a todas las respuestas analizadas. Por su parte, el tercer y cuarto capítulo de esta tesis doctoral se centran en el estudio de las vesículas extracelulares (VEs) como estructuras complejas con potencial participación en los procesos de comunicación entre células de levadura en condiciones de fermentación enológica. Concretamente, en el tercer capítulo no sólo se evalúa si las levaduras enológicas son capaces de producir VEs en un contexto enológico, sino que también se pretende analizar qué proteínas están mayoritariamente presentes en las fracciones enriquecidas en VEs mediante un análisis proteómico. Dicho estudio ha demostrado, de acuerdo con lo descrito en bibliografía, un enriquecimiento en proteínas relacionadas con la pared celular y la glicolisis. Además de la hipótesis de que las VEs actúan como portadoras de señales en las interacciones entre diferentes especies de levaduras vínicas, en el cuarto capítulo de esta tesis doctoral se evalúa si el contenido de proteínas de las VEs se ve, a su vez, afectado por dichas interacciones. Los resultados muestran un claro impacto del co-cultivo sobre la composición proteica en todos los casos. Además, aunque no se puede concluir fírmemente, sí que existen indicios de que algunas levaduras muestran factores comunes en su respuesta a diferentes competidoras, mientras que otras inducen en otras levaduras una respuesta que muestra algún factor común entre varias de ellas. En conjunto, los resultados mostrados en estos dos últimos capítulos apuntan a un potencial papel de las VEs en la comunicación entre diferentes especies de levaduras enológicas. En términos generales, esta tesis doctoral contribuye no sólo al conocimiento fundamental de las interacciones ecológicas a largo plazo, sino que también permite vislumbrar su potencial aplicabilidad industrial. En este sentido, un conocimiento preciso de los mecanismos de interacción permitiría diseñar nuevos cultivos iniciadores mixtos basados en levaduras Saccharomyces y no-Saccharomyces, así como las condiciones óptimas para su aplicación.Universidad de La Rioja (España)González García, Ramón (null)Morales Calvo, Pilar (null)Tronchoni, Jordi (null)2021text (thesis)application/pdfhttps://dialnet.unirioja.es/servlet/oaites?codigo=296473engLICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. 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