An Examination of the Production of Hydrolytic Enzymes and Toxins by Pathogenic Strains of Candida albicans

Chattaway, F. W.; Odds, Frank C.; Barlow, Aaron

doi:10.1099/00221287-67-3-255

Cited by 79 publications

(29 citation statements)

References 19 publications

(12 reference statements)

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“…7). In a second experiment, C. albicans 1161.5.7.7.5 produced more D-arabitol from [2][3][4][5][6][7][8][9][10][11][12][13] C]glucose than did C. albicans 1161, and the 13 C labeling patterns were the same (data not shown). Thus, the inability to produce ArDH affected neither the ability of C. albicans to synthesize D-arabitol from glucose nor the metabolic pathway by which D-arabitol biosynthesis was accomplished.…”

Section: Resultsmentioning

confidence: 91%

“…In one experiment, C. albicans 1161 and 1161.5.7.7.5 were cultured in uridine-supplemented YNB plus 1% [2][3][4][5][6][7][8][9][10][11][12][13] C]glucose at 30ЊC until they reached mid-log phase (A 600 ϭ 0.94 and 1.01, respectively). Gas chromatographic analyses showed that the C. albicans 1161 and 1161.5.7.7.5 extracts contained, respectively, 255 and 103 g of D-arabitol per ml of culture.…”

Section: Resultsmentioning

confidence: 99%

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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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Section: Resultsmentioning

confidence: 91%

Section: Resultsmentioning

confidence: 99%

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

et al. 1995

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“…In addition there is a requirement for a nitrogen source and for salts. The nitrogen sources used have included various peptones ; L-asparagine, provided that serum albumin was also present (Bernander & Edebo, 1969); serum and serum dialysates (Taschdjian & Kozinn, 1961); mixtures of amino acids (Taschdjian & Kozinn, 1961;Johnson, Guzmon & Agurlera, 1954; Mycelium induction in C. albicans 321 Balish & Phillips, 1969; Dabrowa, 1971;Lee, Buckley & Campbell, 1975); single amino acids such as glycine (De Palma, 1966), proline (Dabrowa, 1971) and L-a-amino-n-butyrate (Mardon, Hurst & Balish, 1971); and ammonium sulphate (Chattaway, Odds & Barlow, 1971;Evans et al, 1974;Marriott, 1975). It is difficult to compare the various findings and to reach any general conclusions because of variations in the other factors mentioned above, differences in the method of assessing the extent of mycelial development and the fact that often only a single strain has been examined.…”

Section: Discussionmentioning

confidence: 99%

Induction of the Mycelial Form of Candida albicans by Hydrolysates of Peptides from Seminal Plasma

Chattaway

O’Reilly

Barlow

et al. 1976

Journal of General Microbiology

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SUMMARYPrevious work led to the separation from seminal plasma of a peptide fraction which promoted a high rate of germination of blastospores of Candida albicans. It has now been shown that an acid hydrolysate of this material is also highly active. A minimal amino-acid mixture consisting of aspartic acid, lysine, histidine, threonine, proline and, p-alanine gave go % germination in 4 h with 17 out of 28 strains examined. Glucose and inorganic phosphate were also required. Phosphate was not required for the activity of the original peptide fraction.

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“…6): PHO84, PHO87, PHO100, and VTC4, which encode an enzyme involved in phosphate transport, an enzyme involved in phosphate uptake, a phosphomonoesterase, and a polyphosphate synthase, respectively. Acid phosphatase activity detectable in intact C. albicans cells was proposed many years ago as one of several hydrolytic enzymes with theoretical potential as a virulence factor (10). An inducible phosphomonoesterase, which can be released from intact cells by treatment with sulfhydryl reagents (18), was one of the first biological examples of a highly mannosylated protein with enzyme activity (39).…”

Section: Vol 8 2009mentioning

confidence: 99%

Property Differences among the Four Major Candida albicans Strain Clades

et al. 2009

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A selection of 43 Candida albicans isolates, chosen to represent the four major strain clades of the species and also intraclade diversity, was screened for their virulence in the murine intravenous challenge model of C. albicans infection, for a range of properties measurable in vitro that might relate to virulence, and for the numbers of midrepeat sequences in genes of the ALS and HYR families. Heterozygosity at the mating type locus and low whole-cell acid phosphatase activity and growth rate at 40°C were found to be significantly positively associated with the most virulent isolates. Acid phosphatase activity and growth in 2 M NaCl were statistically significant variables between clades by univariate analysis. Isolates in different clades also differed significantly in midrepeat sequence alleles of ALS2, ALS4, ALS6, ALS7, ALS9, HYR1, and HYR2. There was no association between the midrepeat alleles of any ALS or HYR gene and the virulence of isolates to mice. Genome-wide transcript profiles of 20 isolates (5 per clade) grown under two conditions showed considerable variation between individual isolates, but only a small number of genes showed statistically significant differential gene expression between clades. Analysis of the expression profiles by overall strain virulence revealed 18 open reading frames differing significantly between isolates of high, intermediate, and low virulence. Four of these genes encoded functions related to phosphate uptake and metabolism. This finding and the significant association between whole-cell acid phosphatase activity and virulence led us to disrupt PHO100, which encodes a predicted periplasmic acid phosphatase. The pho100⌬ mutant was mildly but significantly attenuated in terms of survival curves in the mouse model. The study has extended the range of properties known to differ between C. albicans clades and suggests a possible but minor role of phosphate metabolism in the virulence of the species.

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An Examination of the Production of Hydrolytic Enzymes and Toxins by Pathogenic Strains of Candida albicans

Cited by 79 publications

References 19 publications

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

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

Induction of the Mycelial Form of Candida albicans by Hydrolysates of Peptides from Seminal Plasma

Property Differences among the Four Major Candida albicans Strain Clades

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