13C NMR analysis of production and accumulation of osmoregulatory metabolites in the salt-tolerant yeast Debaryomyces hansenii

Jovall, Per‐Åke; Tunblad-Johansson, Inga; Adler, Lennart

doi:10.1007/bf00248956

Cited by 37 publications

(17 citation statements)

References 18 publications

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“…The intracellular salt concentration is maintained at a lower concentration than in the medium. The main osmoregulator in S. cerevisiae appears to be glycerol, whereas in some other yeasts both glycerol and arabinitol are produced intracellularly under salt stress (Reed et al 1987;Meikle et al 1988;Jovall et al 1990). …”

Section: Osmotic Stressmentioning

confidence: 99%

Physiology of yeasts in relation to biomass yields

Verduyn¹

1992

Quantitative Aspects of Growth and Metabolism of Microorganisms

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The stoichiometric limit to the biomass yield (maximal assimilation of the carbon source) is determined by the amount of CO 2 lost in anabolism and the amount of carbon source required for generation of NADPH. This stoichiometric limit may be reached when yeasts utilize formate as an additional energy source. Factors affecting the biomass yield on single substrates are discussed under the following headings: -Energy requirement for biomass formation (YATP)' YATP depends strongly on the nature of the carbon source. Cell composition. The macroscopic composition of the biomass, and in particular the protein content, has a considerable effect on the ATP requirement for biomass formation. Hence, determination of for instance the protein content of biomass is relevant in studies on bioenergetics. Transport of the carbon source. Active (i.e. energy-requiring) transport, which occurs for a number of sugars and polyols, may contribute significantly to the calculated theoretical ATP requirement for biomass formation. Pia-ratio. The efficiency of mitochondrial energy generation has a strong effect on the cell yield. The Pia-ratio is determined to a major extent by the number of proton-translocating sites in the mitochondrial respiratory chain. Maintenance and environmental factors. Factors such as osmotic stress, heavy metals, oxygen and carbon dioxide pressures, temperature and pH affect the yield of yeasts. Various mechanisms may be involved, often affecting the maintenance energy requirement. Metabolites such as ethanol and weak acids. Ethanol increases the permeability ofthe plasma membrane, whereas weak acids can act as proton conductors. Energy content of the growth substrate. It has often been attempted in the literature to predict the biomass yield by correlating the energy content of the carbon source (represented by the degree of reduction) to the biomass yield or the percentage assimilation of the carbon source. An analysis of biomass yields of Candida uti/is on a large number of carbon sources indicates that the biomass yield is mainly determined by the biochemical pathways leading to biomass formation, rather than by the energy content of the substrate.

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Section: Osmotic Stressmentioning

confidence: 99%

Physiology of yeasts in relation to biomass yields

Verduyn¹

1992

Quantitative Aspects of Growth and Metabolism of Microorganisms

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“…Polyols were reported from eukaryotic algae and fungi (Avron, 1986;Wegmann, 1986;Jovall et al, 1990;Van Eck et al, 1989). Glycine betaine and sugars including sugar derivatives were predominantly found in phototrophic eubacteria (Galinski & Truper, 1982;Mackay et al, 1984).…”

Section: Introductionmentioning

confidence: 99%

The predominant role of recently discovered tetrahydropyrimidines for the osmoadaptation of halophilic eubacteria

Severin

Wohlfarth

Galinski

1992

Journal of General Microbiology

176

144

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The aim of this investigation was to perform an extensive screening using HPLC and I3C-NMR spectroscopy to disclose the spectrum of osmolytes produced by aerobic heterotrophic and anoxygenic phototrophic eubacteria. The most predominant solutes detected within a wide range of marine and halophilic micro-organisms were two recently discovered tetrahydropyrimidines ectoine and hydroxyectoine, which were synthesized in response to osmotic stress.

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“…The osmotolerant yeast species Saccharomyces mellis (25), Zygosaccharomyces rouxii (12), and Debaryomyces hansenii (14) convert glucose to D-ribulose-5-PO 4 via the pentose pathway, dephosphorylate D-ribulose-5-PO 4 , and then reduce D-ribulose to D-arabitol with an NADP-dependent pentitol dehydrogenase. In contrast, another strain of Z. rouxii (4) and the marine fungus Dendryphiella salina (18) convert glucose to D-xylulose-5-PO 4 via the pentose pathway, dephosphorylate D-xylulose-5-PO 4 , and then reduce D-xylulose to D-arabitol with an NAD-dependent pentitol dehydrogenase.…”

mentioning

confidence: 99%

“…Although many groups have investigated the diagnostic implications of D-arabitol production by C. albicans, little is known about the metabolic pathways by which C. albicans synthesizes and utilizes D-arabitol, the functions of D-arabitol The evidence supporting the conclusions that fungi metabolize D-arabitol via the pathways described above is indirect, consisting mostly of 14 C and 13 C metabolic labeling data and the presence in cell extracts of enzymes with the expected catalytic activities. To our knowledge, it has not yet been shown for any fungus that a defined mutation or a specific enzyme inhibitor interferes with either biosynthesis or utilization of D-arabitol.…”

mentioning

confidence: 99%

“…The 13 C nuclear magnetic resonance spectroscopy ( 13 C NMR) method described by Jovall et al (14) was adapted to study the metabolic pathway by which C. albicans synthesizes D-arabitol. The C. albicans strains of interest were cultured to mid-late log phase in uridine-supplemented yeast nitrogen base containing 1% [2-13 C]glucose (99 mol%; Sigma Chemical Co., St. Louis, Mo.…”

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D-arabitol metabolism in Candida albicans: construction and analysis of mutants lacking D-arabitol dehydrogenase

et al. 1995

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Candida albicans produces large amounts of the acyclic pentitol D-arabitol in culture and in infected animals and humans, and most strains also grow on minimal D-arabitol medium. An earlier study showed that the major metabolic precursor of D-arabitol in C. albicans was D-ribulose-5-PO 4 from the pentose pathway, that C. albicans contained an NAD-dependent D-arabitol dehydrogenase (ArDH), and that the ArDH structural gene (ARD) encoded a 31-kDa short-chain dehydrogenase that catalyzed the reaction D-arabitol ؉ NAD ‫>؍<‬ D-ribulose ؉ NADH. In the present study, we disrupted both ARD chromosomal alleles in C. albicans and analyzed the resulting mutants. The ard null mutation was verified by Southern hybridization, and the null mutant's inability to produce ArDH was verified by Western immunoblotting. The ard null mutant grew well on minimal glucose medium, but it was unable to grow on minimal D-arabitol The human pathogen Candida albicans produces large amounts of D-arabitol in culture and in infected mammalian hosts (3,7,15,27), and most strains also grow on minimal The evidence supporting the conclusions that fungi metabolize D-arabitol via the pathways described above is indirect, consisting mostly of 14 C and 13 C metabolic labeling data and the presence in cell extracts of enzymes with the expected catalytic activities. To our knowledge, it has not yet been shown for any fungus that a defined mutation or a specific enzyme inhibitor interferes with either biosynthesis or utilization of D-arabitol. Therefore, we sought to ascertain more directly the metabolic functions of ArDH in C. albicans. Specifically, we tested the hypothesis that ArDH catalyzes the last step in D-arabitol biosynthesis and the first step in D-arabitol utilization by disrupting both chromosomal alleles of ARD and by analyzing the resulting null mutants. MATERIALS AND METHODSStrains, media, and plasmids. C. albicans 1161 (arg4 lys1 ser57 MPA1 ura3 gal1) was obtained from S. Scherer (University of Minnesota) (9) and was cultured in yeast extract-peptone supplemented with 1% glucose (YEPD), 1% D-arabitol (YEP/D-arabitol), 1% D-arabinose (YEP/D-arabinose), or no sugar (YEP) (10) or in 0.67% yeast nitrogen base without amino acids (Difco, Detroit, Mich.) supplemented with arginine, lysine, and serine to which 1% glucose (minimal glucose), 1% D-arabitol (minimal D-arabitol), or 1% D-arabinose (minimal D-arabinose) was added. C. albicans transformants were selected on minimal glucose medium plus 1 M sorbitol, and uracil auxotrophs (Ura Ϫ ) were selected on minimal glucose medium supplemented with 625 mg of 5-fluoroorotic acid (FOA) and 100 mg of uridine per liter (FOA medium). Agar (1.5 to 2%) was included as needed.Escherichia coli strains DH5␣ (Gibco BRL, Gaithersburg, Md.) and JM109 (29) were used as plasmid hosts. Cultures were grown in Luria-Bertani medium (20), supplemented with ampicillin (50 g/ml) and/or agar (2%) as needed. E.

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13C NMR analysis of production and accumulation of osmoregulatory metabolites in the salt-tolerant yeast Debaryomyces hansenii

Cited by 37 publications

References 18 publications

Physiology of yeasts in relation to biomass yields

Physiology of yeasts in relation to biomass yields

The predominant role of recently discovered tetrahydropyrimidines for the osmoadaptation of halophilic eubacteria

D-arabitol metabolism in Candida albicans: construction and analysis of mutants lacking D-arabitol dehydrogenase

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