Expression of the GPD1 and GPP2 orthologues and glycerol retention during growth of Debaryomyces hansenii at high NaCl concentrations

Gori, Klaus; Mortensen, Henrik Dam; Arneborg, Nils; Jespersen, Lene

doi:10.1002/yea.1306

Cited by 27 publications

(18 citation statements)

References 35 publications

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“…This finding underlines that microorganisms selected as starter cultures expressing defined functions often behave differently in complex microbial communities or often cannot compete with the indigenous microbiota. Several studies indicate that the microbial succession during cheese ripening depends on adaption of the cultures to stress conditions such as high NaCl concentrations, low water activity [22, 40, 46]. Most recently, growth of aerobic ripening bacteria at cheese surfaces has been shown to be limited by the availability of iron [39].…”

Section: Discussionmentioning

confidence: 99%

Isolation and Identification of the Microbiota of Danish Farmhouse and Industrially Produced Surface-Ripened Cheeses

et al. 2012

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For studying the microbiota of four Danish surface-ripened cheeses produced at three farmhouses and one industrial dairy, both a culture-dependent and culture-independent approach were used. After dereplication of the initial set of 433 isolates by (GTG)5-PCR fingerprinting, 217 bacterial and 25 yeast isolates were identified by sequencing of the 16S rRNA gene or the D1/D2 domain of the 26S rRNA gene, respectively. At the end of ripening, the cheese core microbiota of the farmhouse cheeses consisted of the mesophilic lactic acid bacteria (LAB) starter cultures Lactococcus lactis subsp. lactis and Leuconostoc mesenteorides as well as non-starter LAB including different Lactobacillus spp. The cheese from the industrial dairy was almost exclusively dominated by Lb. paracasei. The surface bacterial microbiota of all four cheeses were dominated by Corynebacterium spp. and/or Brachybacterium spp. Brevibacterium spp. was found to be subdominant compared to other bacteria on the farmhouse cheeses, and no Brevibacterium spp. was found on the cheese from the industrial dairy, even though B. linens was used as surface-ripening culture. Moreover, Gram-negative bacteria identified as Alcalignes faecalis and Proteus vulgaris were found on one of the farmhouse cheeses. The surface yeast microbiota consisted primarily of one dominating species for each cheese. For the farmhouse cheeses, the dominant yeast species were Yarrowia lipolytica, Geotrichum spp. and Debaryomyces hansenii, respectively, and for the cheese from the industrial dairy, D. hansenii was the dominant yeast species. Additionally, denaturing gradient gel electrophoresis (DGGE) analysis revealed that Streptococcus thermophilus was present in the farmhouse raw milk cheese analysed in this study. Furthermore, DGGE bands corresponding to Vagococcus carniphilus, Psychrobacter spp. and Lb. curvatus on the cheese surfaces indicated that these bacterial species may play a role in cheese ripening.

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Section: Discussionmentioning

confidence: 99%

Isolation and Identification of the Microbiota of Danish Farmhouse and Industrially Produced Surface-Ripened Cheeses

et al. 2012

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“…(mg dry weight) −1 . Determination of dry weight was performed as described previously (Gori et al , 2005). Points were fitted to a third‐degree polynomial, and incorporation rates were determined as the differential quotients at selected points.…”

Section: Methodsmentioning

confidence: 99%

Proteomic changes inDebaryomyces hanseniiupon exposure to NaCl stress

Gori

Hébraud²,

Chambon³

et al. 2007

FEMS Yeast Research

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The proteome of the highly NaCl-tolerant yeast Debaryomyces hansenii was investigated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE), and 47 protein spots were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) followed by mass spectrometry (MS). The influence of NaCl on the D. hansenii proteome was investigated during the first 3 h of NaCl exposure. The rate of protein synthesis was strongly decreased by exposure to 8% and 12% (w/v) NaCl, as the average incorporation rates of l-[(35)S]methionine within the first 30 min after addition of NaCl were only 7% and 4% of the rate in medium without NaCl. In addition, the number of protein spots detected on 2D gels prepared from cells exposed to 8% and 12% (w/v) NaCl exceeded less than 28% of the number of protein spots detected on 2D gels prepared from cells without added NaCl. Several proteins were identified as being either induced or repressed upon NaCl exposure. The induced proteins were enzymes involved in glycerol synthesis/dissimilation and the upper part of glycolysis, whereas the repressed proteins were enzymes involved in the lower part of glycolysis, the route to the Krebs cycle, and the synthesis of amino acids. Furthermore, one heat shock protein (Ssa1p) was induced, whereas others (Ssb2p and Hsp60p) were repressed.

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“…This increased expression results in increased synthesis of glycerol, an inert osmolyte that counteracts the effect of extracellular hyperosmolarity (Rep et al , 1999; Estruch, 2000; Gonzalez‐Hernandez and Pena, 2002). Increases in the activity of GPD1 in S. cerevisiae has been well documented by both Northern blots and expression arrays (Rep et al , 2000; Gori et al , 2005; Hirasawa et al , 2006; Causton et al , 2001).…”

Section: Discussionmentioning

confidence: 94%

“…For these experiments, D. hansenii cultures were exposed to 2 M NaCl and were harvested at 30 min, 3 h or 6 h. Control cells were harvested without any prior treatment (termed time 0 control). The three time points were chosen based on Gori et al (2005) (3 h) and Thome and Trench (1999) (0.5 h). We added a 6 h time to differentiate between transiently vs. stably induced genes.…”

Section: Resultsmentioning

confidence: 99%

Genome‐wide expression profiling of the osmoadaptation response of Debaryomyces hansenii

González

Vázquez

Ortíz-Zuazaga

et al. 2009

Yeast

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The euryhaline marine yeast Debaromyces hansenii is a model system for the study of processes related to osmoadaptation. In this study, microarray-based gene expression analyses of the entire genome of D. hansenii was used to study its response to osmotic stress. Differential gene expression, compared to control, was examined at three time points (0.5, 3 and 6 h) after exposure of D. hansenii cultures to high salt concentration. Among the 1.72% of genes showing statistically significant differences in expression, only 65 genes displayed at least three-fold increases in mRNA levels after treatment with 2 M NaCl. On the other hand, 44 genes showed three-fold repression. Upregulated as well as the downregulated genes were grouped into functional categories to identify biochemical processes possibly affected by osmotic stress and involved in osmoadaptation. The observation that only a limited number of genes are upregulated in D. hansenii in response to osmotic stress supports the notion that D. hansenii is pre-adapted to survive in extreme saline environments. In addition, since more than one-half of the upregulated genes encode for ribosomal proteins, it is possible that a translational gene regulatory mechanism plays a key role in D. hansenii's osmoregulatory response. Validation studies for ENA1 and for hyphal wall/cell elongation protein genes, using real-time PCR, confirmed patterns of gene expression observed in our microarray experiments. To our knowledge, this study is the first of its kind in this organism and provides the foundation for future molecular studies assessing the significance of the genes identified here in D. hansenii's osmoadaptation.

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Expression of the GPD1 and GPP2 orthologues and glycerol retention during growth of Debaryomyces hansenii at high NaCl concentrations

Cited by 27 publications

References 35 publications

Isolation and Identification of the Microbiota of Danish Farmhouse and Industrially Produced Surface-Ripened Cheeses

Isolation and Identification of the Microbiota of Danish Farmhouse and Industrially Produced Surface-Ripened Cheeses

Proteomic changes inDebaryomyces hanseniiupon exposure to NaCl stress

Genome‐wide expression profiling of the osmoadaptation response of Debaryomyces hansenii

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