Bases de la respuesta a estrés salino en la levadura debaryomyces hansenii

Resumen

1. Introducción o motivación de la tesis: La mayoría de los seres vivos son sensibles a altas concentraciones de sal, sin embargo, la capacidad de resistir e incluso proliferar en ambientes con baja actividad hídrica debido a una alta concentración de sal en el medio se encuentra ampliamente distribuida en los microorganismos como son, las bacterias, las arqueas y levaduras. En estos grupos es frecuente encontrar especies halotolerantes o halófilas. La necesidad y el interés en la investigación de los efectos del estrés salino y de mecanismos de respuesta se ha incrementado a lo largo de los años. Las levaduras han sido utilizadas como modelo de estudio en estas investigaciones, especialmente, la levadura Saccharomyces cerevisiae. Sin embargo, cada vez se opta más por utilizar las levaduras conocidas como no convencionales, entre ellas Debaryomyces hansenii, debido, por una parte, a las características específicas de estas levaduras y, por otra, por el desarrollo de herramientas moleculares que han propiciado el aprovechamiento del potencial biotecnológico de las levaduras pertenecientes a este grupo. Debaryomyces hansenii es una levadura osmo-, xero- y generalmente considera halotolerante, aunque hay autores que la describen como halófila, que se ha ido utilizado cada vez más frecuentemente como organismo modelo en estudios de estrés salino. Esta levadura posee una serie de adaptaciones que, a pesar de estar presente en la mayoría de las levaduras, proporcionan a Debaryomyces hansenii una mayor tolerancia a la sal que, por ejemplo, a S. cerevisiae. Por otra parte, el metabolismo y el efecto de sal sobre este se encuentra poco estudiado en esta levadura y, además, no es posible obtener conclusiones claras de los pocos estudios realizados debido a las múltiples contradicciones entre los diferentes autores derivadas de la poca homogeneidad de los ensayos realizados. Entre los autores cabe destacar los trabajos del Dr. Antonio Peña de la Universidad de Méjico, en los que se propone que Debaryomyces hansenii posee una capacidad fermentativa y la activación del ciclo del ácido glioxílico en presencia de sal. Este último hecho no ha sido estudiado por autores posteriores. La respuesta al estrés salino constituye un sistema complejo en el que varios elementos se ven implicados, a pesar de los recientes avances realizados en la adaptación de la metodología, y de los resultados obtenidos en los últimos estudios, todavía quedan aspectos por conocer del efecto de las diferentes sales sobre D. hansenii. 2.Contenido de la investigación: En este trabajo se pretende obtener información sobre los diferentes factores implicados en la respuesta a estrés salino en Debaryomyces hansenii, con esta finalidad, en la presente tesis doctoral, se ha abarcado los siguientes contenidos: 1. Se ha determinado el efecto de la salinidad inducida por sodio o litio sobre diferentes procesos fisiológicos y sobre la homeostasis iónica en D. hansenii. 2. Se ha estudiado la influencia del pH, sobre la tolerancia a sodio y litio. 3. Se ha Analizado aspectos básicos del metabolismo de D. hansenii en presencia de sal, como es el consumo de glucosa o la capacidad fermentativa. 4.Se ha determinado cambios en las diferentes rutas metabólicas en respuesta al estrés salino inducido por sodio y litio. 3.Conclusión: 1.El litio, al contrario que el sodio y el potasio, es un elemento tóxico, incluso, en concentraciones muy bajas tanto extracelular como intracelularmente. Sin embargo, no parece producir un daño letal a las células, sino que inhibiría su capacidad de proliferación. 2.La presencia de una concentración de 0,5 M de sodio en el medio favorece el crecimiento de la levadura. Sin embargo, concentraciones más altas de este catión son perjudiciales para la levadura. 3.El sodio, al igual que el potasio, ejerce un efecto protector frente a la toxicidad del litio. 4.El pH no afecta de forma importante al efecto beneficioso de sodio y la toxicidad del litio. 5.La capacidad fermentativa de la cepa CBS767 de D. hansenii es nula tanto en presencia como ausencia de sal en el medio. 6.El sodio influye negativamente a la actividad de enzimas relacionadas con las primeras etapas de la glucólisis, mientras que, una concentración relativamente baja de litio afecta de forma positiva a estas enzimas. 7.La presencia de sodio en el medio induce un cambio en el patrón metabólico a favor del ciclo del ácido glioxílico. 8.Al contrario que el sodio, el litio favorece el flujo metabólico hacía el ciclo de Krebs. 4. Bibliografía: - Adler, L., Blomberg, A., & Nilsson, A. (1985). Glycerol metabolism and osmoregulation in the salt-tolerant yeast Debaryomyces hansenii. 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