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PubMed2006

The effect of citric acid and pH on growth and metabolism of anaerobic Saccharomyces cerevisiae and Zygosaccharomyces bailii cultures.

The effects of citric acid at pH values of 3.0, 4.0, and 4.5 on growth and metabolism of anaerobic Saccharomyces cerevisiae and Zygosaccharomyces bailii cultures were investigated. S. cerevisiae and Z. bailii exhibited similar tolerances to citric acid, as determined by growth measurements, at all three pH values investigated. The citric-acid-induced growth inhibition of both yeast species increased with increasing pH values, indicating that the antimicrobial mechanism of citric acid differs from that of classical weak-acid preservatives. In S. cerevisiae, citric acid shifted the primary energy metabolism towards lower ethanol production and higher glycerol production, thus resulting in lower ATP production. These metabolic changes in S. cerevisiae were pH-dependent; i.e. the higher the pH, the lower the ATP production, and they may explain why growth of S. cerevisiae is more inhibited by citric acid at higher pH values. In Z. bailii, citric acid also caused an increased glycerol production, although to a lesser extent than in S. cerevisiae, but it caused virtually no changes in ethanol and ATP production.

Adenosine Triphosphate↗
PubMed1988

Clinical evaluation of the use of citric acid and autologous fibronectin in periodontal surgery.

This study evaluated the effects of citric acid demineralization and autologous fibronectin application in association with a modified Widman flap in the treatment of periodontitis. The study population comprised 29 patients under treatment for moderate to advanced periodontitis who reached the one-year posttherapy evaluation. After thorough scaling and root planing, a split mouth design was used in which two quadrants were treated by modified Widman flap alone, and the other two randomly assigned quadrants were treated by modified Widman flap combined with citric acid demineralization and autologous fibronectin application. Fibronectin, which had previously been isolated from the patient's own plasma, was applied with a tuberculin syringe on the citric acid demineralized root surfaces and the inner aspect of the flap. After suturing provided good flap adaptation, additional fibronectin was again applied under the flap and external pressure was applied. Patients were clinically evaluated at baseline and at one year. Statistical evaluation of the data using paired t test and Chi-square analysis indicated that both approaches, modified Widman flap alone or in combination with citric acid and fibronectin, significantly reduced probing pocket depth and increased clinical attachment. However, the changes achieved with citric acid and fibronectin were statistically greater than those obtained with the flap alone. Furthermore, the number of sites gaining 2 mm or more of clinical attachment were significantly increased. The results suggest that the use of citric acid and fibronectin holds promise in promoting reattachment after periodontal therapy.

Adult↗
PubMed2002

Citric acid production by Candida strains under intracellular nitrogen limitation.

A suitable strain and important factors influencing citric acid formation in yeasts were identified. Candida oleophila ATCC 20177 was chosen as the best citric acid producer from several Candida strains. Yields of 50 g/l citric acid were produced in shake flask and 80 g/l in fed-batch fermentations with 1.5 and 3 g/l NH(4)Cl under non-optimized conditions. Ammonium nitrogen was identified as the limiting substrate for citrate formation. Citric acid excretion begins a few hours after exhaustion of nitrogen in the medium. The importance of intracellular nitrogen limitation was clarified by elemental analysis of C. oleophila biomass. The nitrogen content of C. oleophila biomass decreased from 7.45% during the growth phase to 3.96% in the production phase. The biomass contained less carbon and more trace elements in the growth phase compared with the production phase. Relatively high intracellular NH(4)(+) concentration of about 1.2 mg/g biomass (~37.4 mM) was found during the production phase. The low intracellular nitrogen content and increase of intracellular NH(4)(+) concentration, possibly caused by proteolysis following extracellular nitrogen exhaustion, trigger citric acid production. Intracellular nitrogen limitation and the increase in intracellular NH(4)(+) concentration are the most important factors influencing citric acid formation in yeasts.

Biomass↗
PubMed2005

Combined efficacy of nisin and pediocin with sodium lactate, citric acid, phytic acid, and potassium sorbate and EDTA in reducing the Listeria monocytogenes population of inoculated fresh-cut produce.

The inability of chlorine to completely inactivate human bacterial pathogens on whole and fresh-cut produce suggests a need for other antimicrobial washing treatments. Nisin (50 microg/ml) and pediocin (100 AU/ml) individually or in combination with sodium lactate (2%), potassium sorbate (0.02%), phytic acid (0.02%), and citric acid (10 mM) were tested as possible sanitizer treatments for reducing the population of Listeria monocytogenes on cabbage, broccoli, and mung bean sprouts. Cabbage, broccoli, and mung bean sprouts were inoculated with a five-strain cocktail of L. monocytogenes at 4.61, 4.34, and 4.67 log CFU/g, respectively. Inoculated produce was left at room temperature (25 degrees C) for up to 4 h before antimicrobial treatment. Washing treatments were applied to inoculated produce for 1 min, and surviving bacterial populations were determined. When tested alone, all compounds resulted in 2.20- to 4.35-log reductions of L. monocytogenes on mung bean, cabbage, and broccoli, respectively. The combination treatments nisin-phytic acid and nisin-pediocin-phytic acid caused significant (P < 0.05) reductions of L. monocytogenes on cabbage and broccoli but not on mung bean sprouts. Pediocin treatment alone or in combination with any of the organic acid tested was more effective in reducing L. monocytogenes populations than the nisin treatment alone. Although none of the combination treatments completely eliminated the pathogen on the produce, the results suggest that some of the treatments evaluated in this study can be used to improve the microbial safety of fresh-cut cabbage, broccoli, and mung bean sprouts.

Anti-Bacterial Agents↗
PubMed2004

The effect of EDTA and citric acid on phytoremediation of Cd, Cr, and Ni from soil using Helianthus annuus.

The possibility to clean heavy metal contaminated soils with hyperaccumulator plants has shown great potential. One of the most recently studied species used in phytoremediation applications are sunflowers. In this study, two cultivars of Helianthus annuus were used in conjunction with ethylene diamine tetracetic acid (EDTA) and citric acid (CA) as chelators. Two different concentrations of the chelators were studied for enhancing the uptake and translocation of Cd, Cr, and Ni from a silty-clay loam soil. When 1.0 g/kg CA was used, the highest total metal uptake was only 0.65 mg. Increasing the CA concentration posed a severe phytotoxicity to both cultivars as evidenced by stunted growth and diminished uptake rates. Decreasing the CA concentration to 0.1 and 0.3 g/kg yielded results that were not statistically different from the control. EDTA at a concentration of 0.1 g/kg yielded the best results for both cultivars achieving a total metal uptake of approximately 0.73 mg compared to approximately 0.40 mg when EDTA was present at 0.3 g/kg.

Biodegradation, Environmental↗
PubMed1999

The effects of extracellular citric acid acidosis on the viability, cellular adhesion capacity and protein synthesis of cultured human gingival fibroblasts.

Root surface demineralization is widely used as an adjunct to periodontal treatment. To clarify the influence of citric acid root conditioning on periodontal wound healing, the effects of citric acid and associated extracellular acidosis on the viability (MTT assay), attachment and protein synthesis ([3H]-proline incorporation into trichloroacetic acid-precipitated proteins) of human gingival fibroblasts (GF) were investigated. A concentration of 47.6 mmol/L of citric acid (pH 2.3) in water led to total cell death within three minutes of incubation. Media containing 23.8 mmol/L and 47.6 mmol/L of citric acid exerted strong cytotoxicity (47 to 90 per cent of cell death) and inhibited protein synthesis (IC50 = 0.28 per cent) of GF within three hours of incubation. Incubation of cells in a medium containing 11.9 mmol/L of citric acid also suppressed the attachment and spreading of fibroblasts on culture plates and Type I collagen, with 58 per cent and 22 per cent of inhibition, respectively. Culture medium supplemented with 11.9, 23.8 and 47.6 mmol/L of citric acid also led to extracellular acidosis by decreasing the pH value from 7.5 to 6.3, 5.2 and 3.8, respectively. In addition, it was confirmed that the toxic effect of media containing citric acid was due to their acidity rather than the citrate content. Most of the citric acid-induced cell death could be prevented by adjusting the pH value of the culture medium to pH 7.5. Sodium citrate, at a concentration of 47.6 mmol/L, also exerted little cytotoxicity. The results suggested that toxicity of citric acid in specific stages of the healing process must be considered prior to its clinical application. Careful management of citric acid in order to avoid contact with tissue or the development of other demineralizing agents is important in enhancing periodontal wound healing.

Acidosis↗
PubMed1982

Anti-bacterial effect of citric acid treatment of periodontally diseased root surfaces in vitro.

This investigation examined whether citric acid may exert an anti-bacterial effect against plaque deposits on root surfaces in vitro. Aerobic and anaerobic blood-agar plate cultures were prepared from plaque samples obtained from the proximal root surfaces of 20 periodontally diseased human teeth following extraction. Ten teeth were exposed to saturated citric acid (pH 1) for 3 min, followed by rinsing in sterile 0.85% saline and plaque samples were then obtained immediately adjacent to those sites sampled initially. Controls consisted of using sterile water instead of citric acid on a further five teeth. The numbers of colonies present on pre- and post-treatment culture plates were counted at 24 h. The results indicated that citric acid application reduced, in all instances, the numbers of colonies grown from post-exposure plaque samples as compared to pre-exposure samples. No colonies were detected in 55% of aerobic and 30% of anaerobic cultures of acid-treated root surface samples. For aerobic cultures, citric acid exposure reduced the number of colonies grown from greater than 10(4) to less than 100 in 95% of the root surfaces sampled, while for anaerobic cultures, reduction from greater than 10(4) to less than 100 was found in 80% of surfaces sampled. The findings indicate that citric acid exerts anti-bacterial activity against microbial plaque deposits present on periodontally diseased root surfaces in vitro.

Citrates↗
PubMed1982

The interrelationships between acid phosphatase, aminopeptidase, diamine oxidase, citric acid, beta-glucuronidase, pH and zinc in human prostatic fluid.

Biochemical analysis of 328 human prostatic fluid samples were performed. The urea concentration of 69 samples was similar to that of serum and not indicative of significant contamination with urine. The pH of normal fluids was acidic (mean pH = 6.7). The interrelationships between zinc, citrate, acid phosphatase aminopeptidase, beta-glucuronidase, diamine oxidase and pH were investigated by factor analysis. Two significant factors were extracted, the first accounted for 89% of their common variance and the second for 11%. Zinc, citrate, acid phosphatase and aminopeptidase were positively and pH was negatively related to factor one. Beta-glucuronidase was positively related to factor two and diamine oxidase was largely independent of both factors. It was concluded that variables related to factor one share a common secretory control and mechanism, that some other mechanism operates in the case of beta-glucuronidase and that diamine oxidase may not be a true secretory product of the prostate.

Acid Phosphatase↗
PubMed1987

Organizational aspects of the citric acid cycle.

The enzymes of the citric acid cycle show at least two levels of organization within the mitochondrial matrix. Six of the possible eight sequential enzymes show specific interactions in vitro. Further, the enzymes bind specifically to the matrix surface of the inner membrane. A slightly damaged mitochondrial particle has been isolated which contains bound, but exposed, Krebs citric acid cycle enzymes. This particle (a metabolon) shows a kinetic advantage for two coupled systems, fumarate oxidation and malate conversion to citrate, over a solubilized system. N.m.r. experiments indicate that many components of the matrix are in a bound state.

Animals↗
PubMed1986

Influence of ingestion of aluminum, citric acid and soil on mineral metabolism of lactating beef cows.

Lactating beef cows (16 Hereford and 34 Angus, 430 kg average body weight, aged 8 to 10 yr) were fed a basal diet containing 200 micrograms/g Al alone or supplemented with Al-citrate, citric acid, soil or soil plus citric acid for 56 d. Diets containing Al-citrate, soil and soil plus citric acid contained 1,730, 1,870 and 1,935 micrograms/g Al, dry-basis, respectively. Adding soil to the diet also increased Mg and Fe content of the diet. Aluminum values in ruminal contents of beef cows fed the basal alone or supplemented with citric acid, Al-citrate, soil or soil plus citric acid were 800, 990, 2,930, 3,410 and 2,910 micrograms/g, air-dry basis, respectively. Serum Mg and inorganic P declined (P less than .01) and urinary Ca concentration increased (P less than .01) for cows fed Al-citrate. By d 56, serum Mg was 1.5 and 2.2 mg/dl, and serum P was 3.8 and 6.8 mg/dl, for cows fed Al-citrate and basal diets, respectively. Calcium concentrations in urine were 281 and 11 micrograms/g for cows fed Al-citrate and basal diets, respectively. Citric acid, soil and soil plus citric acid had no detrimental effects on serum Mg and inorganic P, or urinary Ca concentration. By d 56, serum Ca was higher (P less than .06) in cows fed Al-citrate, compared with cows on the other four diets. Bone Ca, P, Zn and percent ash were not significantly affected by treatment but bone Mg tended to be slightly lower (P less than .07) for cows fed Al-citrate.(ABSTRACT TRUNCATED AT 250 WORDS)

Aluminum↗
PubMed2003

Do mitochondria regulate cellular iron homeostasis through citric acid and haem production? Implications for cancer and other diseases.

Citric acid is produced industrially by depriving Aspergillus niger of iron. The lack of Fe deactivates mitochondrial aconitase and interrupts the krebs cycle, causing the mitochondria to release citric acid as a siderophore (an Fe getter). When the mitochondrion has plenty of Fe and the cell has enough ATP, aerobic phosphorylation stops and fatty acid or haem synthesis take place, when the cell has plenty of haem, haem synthesis stops. Since most of the Fe activity in the cell is related to the mitochondria, I hypothesise that in the animal cell when the mitochondria are low in Fe, citric acid acts as a signal that triggers the production of transferrin receptor messenger RNA (TrR mRNA) in the nucleus, which in the absence of Fe causes the expression of transferrin receptor. When the cell has plenty of Fe, cytosolic aconitase detaches itself from the transferrin receptor and ferritin mRNA stopping expression of the former and initiating expression of the latter. The detached cytosolic aconitase transforms the citric acid, blocking the production of the transferrin receptor mRNA.

Blood Platelets↗
PubMed2004

Citric acid cycle intermediates as ligands for orphan G-protein-coupled receptors.

The citric acid cycle is central to the regulation of energy homeostasis and cell metabolism. Mutations in enzymes that catalyse steps in the citric acid cycle result in human diseases with various clinical presentations. The intermediates of the citric acid cycle are present at micromolar concentration in blood and are regulated by respiration, metabolism and renal reabsorption/extrusion. Here we show that GPR91 (ref. 3), a previously orphan G-protein-coupled receptor (GPCR), functions as a receptor for the citric acid cycle intermediate succinate. We also report that GPR99 (ref. 4), a close relative of GPR91, responds to alpha-ketoglutarate, another intermediate in the citric acid cycle. Thus by acting as ligands for GPCRs, succinate and alpha-ketoglutarate are found to have unexpected signalling functions beyond their traditional roles. Furthermore, we show that succinate increases blood pressure in animals. The succinate-induced hypertensive effect involves the renin-angiotensin system and is abolished in GPR91-deficient mice. Our results indicate a possible role for GPR91 in renovascular hypertension, a disease closely linked to atherosclerosis, diabetes and renal failure.

Animals↗
PubMed1985

Diurnal variation and adaptation of the cough response to citric acid in normal subjects.

The lowest concentrations of citric acid were measured that caused coughing in 10 normal subjects who inhaled successively higher concentrations. Two subjects did not cough at any concentration. In the remaining eight the threshold concentration was significantly higher when measured in the afternoon than it was in the morning (p less than 0.05). The expected diurnal variation in peak expiratory flow rate (PEFR) was found (significant in the whole group--p less than 0.05); but PEFR did not change significantly when measured before and immediately after coughing caused by citric acid inhalations. In a second group of 10 normal subjects two series of citric acid inhalations were given, separated by one hour. The total number of coughs was significantly lower on the second run (p less than 0.05). Thus diurnal variation and adaptation of the cough response must be taken into account when antitussive drugs are tested.

Adaptation, Physiological↗
PubMedDate not supplied

Chemical evolution of the citric acid cycle: sunlight photolysis of the amino acids glutamate and aspartate.

Sunlight photolysis of the amino acids glutamate and aspartate were carried out on 0.1 M aqueous solutions at pH = 7.0. The non-volatile products were identified by GC-MS analysis of derived methyl esters. The major product from glutamic acid was succinic acid, and, analogously, aspartic acid photolyzed to malonic acid. The photochemical oxidative decarboxylation of glutamate parallels its metabolism in modern cells and may provide an evolutionary link between simple amino acids and reactions of the citric acid cycle.

Aspartic Acid↗
PubMed2003

Stimulatory effect of alcohols (methanol and ethanol) on citric acid productivity by a 2-deoxy D-glucose resistant culture of aspergillus niger GCB-47.

The present study describes citric acid fermentation by Aspergillus niger GCB-47 in a 15-1 stainless steel stirred fermentor. Among the alcohols tested as stimulating agents, 1.0% (v/v) methanol was found to give maximum amount of anhydrous citric acid (90.02 +/- 2.2 g/l), 24 h after inoculation. This yield of citric acid was 1.96 fold higher than the control. Methanol has a direct effect on mycelial morphology and it promotes pellet formation. It also increases the cell membrane permeability to provoke more citric acid excretion from the mycelial cells. The sugar consumed and % citric acid was 108 +/- 3.8 g/l and 80.39 +/- 4.5%, respectively. The desirable mycelial morphology was in the form of small round pellets having dry cell mass 14.5 +/- 0.8 g/l. Addition of ethanol, however, did not found to enhance citric acid production, significantly. The maximum value of Yp/x (i.e., 5.825 +/- 0.25 g/g) was observed when methanol was used as a stimulating agent. The best results of anhydrous citric acid were observed, 6 days after inoculation when the initial pH of fermentation medium was kept at 6.0.

Aspergillus niger↗
PubMed2001

Optimization of citric acid production from Candida lipolytica Y-1095 using n-paraffin.

Currently, the majority of worldwide microbial production of citric acid utilizes Aspergillus niger in a carbohydrate based submerged fermentation. Due to their high carbon content, hydrocarbons also have the potential of producing high concentrations of citric acid. Initial lab experiments conducted using 1875 ml batch fermentations with n-paraffin found that Candida lipolytica NRRL-Y-1095 assimilated the feedstock and had a citric acid productivity of 47 mg l(-1) h(-1). To determine the optimum level of initial biomass concentration, n-paraffin concentration, iron concentration and temperature for the production of citric acid, a central composite design was developed using 200 ml batch fermentations. The design involved conducting 31 batch fermentations under various combinations of high and low values of these four parameters. From this investigation empirical models were developed describing the interactions between the experimental parameters and citric acid production. It was found that the maximum concentration of citric acid produced was 9.8 g l(-1) and the optimum levels of each parameter for citric acid production were, 10--12% volume for initial biomass concentration, 10--15% volume for n-paraffin concentration, 10 mg l(-1) for ferric nitrate concentration, and 26--30 degrees C for temperature.

Candida↗
PubMed2003

Modeling the adsorption of citric acid onto Muloorina illite and related clay minerals.

The adsorption of citric acid onto goethite, kaolinite, and illite was measured as a function of pH (adsorption edges) and concentration (adsorption isotherms) at 25 degrees C. The greatest adsorption was onto goethite and the least onto illite. Adsorption onto goethite was at a maximum below pH 5 and decreased as the pH was increased to pH 9. For kaolinite, maximum adsorption occurred between pH 4.5 and pH 7, decreasing below and above this pH region, while for illite maximum adsorption occurred between about pH 5 and pH 7, decreasing at both lower and higher pH. ATR-FTIR spectra of citrate adsorbed to goethite at pH 4.6, pH 7.0, and pH 8.8 were compared with those of citrate solutions between pH 3.5 and pH 9.1. While the spectra of adsorbed citrate resembled those of the fully deprotonated solution species, there were significant differences. In particular the C[bond]O symmetric stretching band of the adsorbed species at pH 4.6 and 7.0 changed shape and was shifted to higher wave number. Further spectral analysis suggested that citrate adsorbed as an inner-sphere complex at pH 4.6 and pH 7.0 with coordination to the surface most probably via one or more carboxyl groups. At pH 8.8 the intensity of the adsorbed bands was much smaller but their shape was similar to those from the deprotonated citrate solution species, suggesting outer-sphere adsorption. Insufficient citric acid adsorbed onto illite or kaolinite to provide spectroscopic information about the mode of adsorption onto these minerals. Data from adsorption experiments, and from potentiometric titrations of suspensions of the minerals in the presence of citric acid, were fitted by extended constant-capacitance surface complexation models. On the goethite surface a monodentate inner-sphere complex dominated adsorption below pH 7.9, with a bidentate outer-sphere complex required at higher pH values. On kaolinite, citric acid adsorption was modeled with a bidentate outer-sphere complex at low pH and a monodentate outer-sphere complex at higher pH. There is evidence of dissolution of kaolinite in the presence of citric acid. For illite two bidentate outer-sphere complexes provided a good fit to all data.

Journal Article↗