RESUMO
Debaryomyces hansenii is a halotolerant yeast of importance in basic and applied research. Previous reports hinted about possible links between saline and oxidative stress responses in this yeast. The aim of this work was to study that hypothesis at different molecular levels, investigating after oxidative and saline stress: (i) transcription of seven genes related to oxidative and/or saline responses, (ii) activity of two main anti-oxidative enzymes, (iii) existence of common metabolic intermediates, and (iv) generation of damages to biomolecules as lipids and proteins. Our results showed how expression of genes related to oxidative stress was induced by exposure to NaCl and KCl, and, vice versa, transcription of some genes related to osmotic/salt stress responses was regulated by H2O2. Moreover, and contrary to S. cerevisiae, in D. hansenii HOG1 and MSN2 genes were modulated by stress at their transcriptional level. At the enzymatic level, saline stress also induced antioxidative enzymatic defenses as catalase and glutathione reductase. Furthermore, we demonstrated that both stresses are connected by the generation of intracellular ROS, and that hydrogen peroxide can affect the accumulation of in-cell sodium. On the other hand, no significant alterations in lipid oxidation or total glutathione content were observed upon exposure to both stresses tested. The results described in this work could help to understand the responses to both stressors, and to improve the biotechnological potential of D. hansenni.
Assuntos
Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Estresse Oxidativo/fisiologia , Saccharomycetales/fisiologia , Estresse Salino/fisiologia , Antioxidantes , Catalase/metabolismo , Proteínas de Ligação a DNA/genética , Regulação Fúngica da Expressão Gênica , Genes Fúngicos/genética , Glutationa/metabolismo , Glutationa Redutase/metabolismo , Peróxido de Hidrogênio , Metabolismo dos Lipídeos , Osmorregulação/genética , Osmorregulação/fisiologia , Estresse Oxidativo/genética , Cloreto de Potássio/metabolismo , Proteômica , Saccharomycetales/genética , Estresse Salino/genética , Sódio/metabolismo , Cloreto de Sódio/metabolismo , Fatores de Transcrição/genéticaRESUMO
Yeasts, filamentous fungi, and bacteria colonize the surface of fermented sausages during the ripening process. The source of this microbiota is their surrounding environment, and is influenced by the maturing conditions and starter cultures. Debaryomyces hansenii was previously isolated from several dry-cured meat products and associated with the lipolytic and proteolytic changes that occur in these products, influencing their taste and flavor. Therefore, this study isolated the yeast microbiota present in the casing from different meat products ("lomo," "chorizo," and "salchichón") from the Valle de los Pedroches region in southern Spain. D. hansenii was by far the most abundant species in each product, as all 22 selected isolates were identified as D. hansenii by biochemical and/or molecular methods. In contrast, no yeasts were found in the meat batter. These data constitute the first study of the yeasts present in "lomo" sausages and particularly the highly appreciated Valle de los Pedroches "lomo" sausages. Furthermore, the resistance of these isolates to different pHs, temperatures, and saline stress was studied, together with their catabolic characteristics. Based on the results, certain isolates are proposed as valuable candidate starter cultures that could improve both the manufacture and the flavor of such dry-cured meat products, and provide an understanding of new mechanisms involved in stress tolerance. Applied mediumscale industrial tests are currently in progress.
Assuntos
Produtos da Carne/microbiologia , Saccharomycetales , Animais , Contagem de Colônia Microbiana , Concentração de Íons de Hidrogênio , Saccharomycetales/genética , Saccharomycetales/isolamento & purificação , Saccharomycetales/metabolismo , Saccharomycetales/fisiologia , Espanha , TemperaturaRESUMO
Debaryomyces hansenii is a halotolerant yeast with a high biotechnological potential, particularly in the food industry. However, research in this yeast is limited by its molecular peculiarities. In this review we summarize the state of the art of research in this microorganisms, describing both pros and cons. We discuss (i) its halotolerance, (ii) the molecular factors involved in saline and osmotic stress, (iii) its high gene density and ambiguous CUG decoding, and (iv) its biotechnological and medical interests. We trust that all the bottlenecks in its study will soon be overcome and D. hansenii will become a fundamental organism for food biotechnological processes. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Biotecnologia , Debaryomyces/fisiologia , Tecnologia de Alimentos/tendências , Biotecnologia/tendências , Cátions/metabolismo , Debaryomyces/genética , Genoma Fúngico/genética , Halogênios/metabolismo , Pressão Osmótica/fisiologiaRESUMO
We report the characterization of five strains belonging to the halotolerant highly related Debaryomyces hansenii/fabryi species. The analysis performed consisted in studying tolerance properties, membrane characteristics, and cation incell amounts. We have specifically investigated (1) tolerance to different chemicals, (2) tolerance to osmotic and salt stress, (3) tolerance and response to oxidative stress, (4) reactive oxygen species (ROS) content, (5) relative membrane potential, (6) cell volume, (7) K(+) and Na(+) ion content, and (8) membrane fluidity. Unexpectedly, no direct relationship was found between one particular strain, Na(+) content and its tolerance to NaCl or between its ROS content and its tolerance to H(2)O(2). Results show that, although in general, human origin D. fabryi strains were more resistant to oxidative stress and presented shorter doubling times and smaller cell volume than food isolated D. hansenii ones, strains belonging to the same species can be significantly different. Debaryomyces fabryi CBS1793 strain highlighted for its extremely tolerant behavior when exposed to the diverse stress factors studied.