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PubMed1986

The characterization and cloning of a gluconate (gnt) operon of Bacillus subtilis.

The enzymes involved in gluconate utilization in Bacillus subtilis seemed to be gluconate permease and gluconate kinase. Several mutants unable to grow on gluconate were isolated. The mutations they harboured (gnt) were clustered between iol-6 and fdp-74 on the B. subtilis chromosome (a tentative map order of gnt-10, gnt-4, gnt-26, gnt-23 and gnt-9 was obtained). The gnt-10 mutation seemed to be located within the structural gene of the kinase, and the gnt-23 and gnt-26 mutations seemed to be within that of the permease. An EcoRI fragment (4.5 MDal) containing an intact gluconate (gnt) operon consisting of these two structural genes was cloned in phage phi 105 by prophage transformation and was mapped physically. The physical location of the mutations coincided with their order on the genetic map. The HindIII-A fragment (2.4 MDal), which corrects all the gnt mutations, was subcloned in plasmid pC194. The fragment contained the structural genes for the gluconate permease and kinase, but not the regulatory region of the gluconate operon.

Bacillus subtilis↗
PubMedDate not supplied

The efficacy of treatment with triamcinolone acetonide in calcinosis cutis following extravasation of calcium gluconate: a preliminary study.

Neonatal hypocalcemia is not an uncommon condition, especially in the premature neonate. It is effectively treated by intravenous administration of calcium gluconate. Complications of extravasation during intravenous infusion include localized calcification and occasionally necrosis. When this occurs, however, there is no specific mode of treatment except supportive management and skin graft. This experiment was designed to evaluate the efficacy and safety of treatment with triamcinolone acetonide in calcinosis cutis following extravasation of calcium gluconate. Initially, 2 cc of 10% calcium gluconate was injected subcutaneously into two rabbits at seven sites on the shaved skin of the back. Another two rabbits were injected at the same sites with 0.5 cc of triamcinolone acetonide (10 mg/dl) after injection of 2 cc of 10% calcium gluconate. As a control, 2 cc of normal saline was injected into another rabbit in the same manner. These five rabbits were observed over the next 7 weeks and underwent pathologic examination at various intervals (on days 1, 3, 8, 15, 30, 37, 45). In the 10% calcium gluconate injected rabbits, nodules and large ulcerated lesions developed after 15 days. Multiple, linear, ulcerative and indurated masses were noted on day 37. The lesions healed progressively with a decrease in ulceration, and after 2 months, the masses disappeared gradually. Histologically, on day 15 calcium deposits were seen in the walls of the arteries, veins, dermis, and muscle fibers and epidermal necrosis was seen at the injection sites. From day 37 discharge of calcium deposits began to take place by means of transepidermal elimination. After 2 months, the calcium and mucin deposition was observed focally in the dermis. In the rabbits injected with 10% calcium gluconate and triamcinolone acetonide, mild erythema and induration were seen after day 15 at the injection sites; this gradually disappeared. After 30 days the injection sites were normal in appearance. Histologically, at day 15 calcium deposition was seen in the upper dermis, but after 1 month the injection sites were histologically normal. We suggest that intralesional injection of triamcinolone acetonide for the treatment of calcinosis cutis following extravasation of calcium gluconate is effective, probably due to its antiinflammatory effect and its role in facilitating the resorption of calcium in the tissue.

Administration, Topical↗
PubMedDate not supplied

Muscle potassium content and potassium gluconate supplementation in normokalemic cats with naturally occurring chronic renal failure.

Muscle potassium content and supplementation with potassium gluconate were evaluated in normokalemic cats with chronic renal failure (CRF). Affected cats received standard medical therapy for renal failure and either placebo (sodium gluconate) or potassium gluconate. At the beginning of the study and after 6 months of supplementation, glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were estimated using 3H-inulin and 14C-tetraethylammonium bromide (TEA) clearances. Muscle potassium content was determined in biopsy specimens using atomic absorption spectroscopy. Muscle biopsy samples obtained from cats with CRF before treatment had significantly lower muscle potassium content than did those from normal control cats. Over the 6-month period of supplementation, muscle potassium content increased both in cats with CRF that received potassium gluconate and in those that received placebo (sodium gluconate). Serum potassium concentration and fractional excretion of potassium remained relatively unchanged in both groups of cats throughout the treatment period. There were no significant differences in the percentage change in GFR and ERPF between treatment groups over the 6-month time period. Median values for pH, HCO3-, and total CO2 at 6 months were higher than baseline in the potassium gluconate group but lower than baseline in the sodium gluconate group.

Animals↗
PubMed1997

Cloning and expression of a gene cluster encoding three subunits of membrane-bound gluconate dehydrogenase from Erwinia cypripedii ATCC 29267 in Escherichia coli.

We have cloned the gene cluster encoding three subunits of membrane-bound gluconate dehydrogenase (GADH) from Erwinia cypripedii ATCC 29267 in Escherichia coli by performing a direct-expression assay. The positive clone converted D-gluconate to 2-keto-D-gluconate (2KDG) in the culture medium. Nucleotide sequence analysis of the GADH clone revealed that the cloned fragment contained the complete structural genes for a 68-kDa dehydrogenase subunit, a 47-kDa cytochrome c subunit, and a 24-kDa subunit of unknown function and that the genes were clustered with the same transcriptional polarity. Comparison of the deduced amino acid sequences and the NH2-terminal sequences determined for the purified protein indicated that the dehydrogenase, cytochrome c, and 24-kDa subunits contained typical signal peptides of 22, 19, and 42 amino acids, respectively. The molecular masses of the processed subunits deduced from the nucleotide sequences (65, 45, and 20 kDa) coincided well with the molecular masses of subunits estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In E. cypripedii and recombinant E. coli, the GADH was constitutively formed and the activity of GADH was enhanced more than twofold by addition of D-gluconate to the medium. The holoenzyme glucose dehydrogenase of E. coli was reconstituted by addition of pyrroloquinoline quinone to the culture medium, and the conversion of D-glucose or D-gluconate to 2KDG by recombinant E. coli harboring the cloned GADH gene was attempted in batch culture. The conversion yields for D-glucose were 0.95 mol of 2KDG/mol of D-glucose after 16 h of cultivation, and those for D-gluconate were 0.95 mol of 2KDG/mol of D-gluconate after 12 h of cultivation.

Amino Acid Sequence↗
PubMed1967

Glucose and gluconate metabolism in an Escherichia coli mutant lacking phosphoglucose isomerase.

A single gene mutant lacking phosphoglucose isomerase (pgi) was selected after ethyl methane sulfonate mutagenesis of Escherichia coli strain K-10. Enzyme assays revealed no pgi activity in the mutant, whereas levels of glucokinase, glucose-6-phosphate dehydrogenase, and gluconate-6-phosphate dehydrogenase were similar in parent and mutant. The amount of glucose released by acid hydrolysis of the mutant cells after growth on gluconate was less than 2% that released from parent cells; when grown in the presence of glucose, mutant and parent cells contained the same amount of glucose residues. The mutant grew on glucose one-third as fast as the parent; it also grew much slower than the parent on galactose, maltose, and lactose. On fructose, gluconate, and other carbon sources, growth was almost normal. In both parent and mutant, gluconokinase and gluconate-6-phosphate dehydrase were present during growth on gluconate but not during growth on glucose. Assay and degradation of alanine from protein hydrolysates after growth on glucose-1-(14)C and gluconate-1-(14)C showed that in the parent strain glucose was metabolized by the glycolytic path and the hexose monophosphate shunt. Gluconate was metabolized by the Entner-Doudoroff path and the hexose monophosphate shunt. The mutant used glucose chiefly by the shunt, but may also have used the Entner-Doudoroff path to a limited extent.

Alanine↗
PubMed1986

Utilization of gluconate by Aspergillus niger. II. Enzymes of degradation pathways and main end products.

Aspergillus niger was grown from conidia on a medium with glucose as the source of carbon and potassium nitrate in non-limiting concentration as the source of nitrogen. After the exhaustion of glucose gluconate was added, this compound representing the almost only carbon source in the culture fluid at this time. Gluconate was used rapidly by the preformed mycelium, the main end-products of its metabolization being mycelial substance (including protein), CO2, and oxalate. In cell-free extracts from gluconate utilizing mycelia 8 enzymes of the Embden-Meyerhof (EM) pathway, 5 enzymes of the tricarboxylic acid (TCA) cycle, and an oxalate forming enzyme, oxaloacetate hydrolase (EC 3.7.1.1) were identified. The addition of fluoroacetate together with gluconate resulted in the accumulation of citrate, and in the inhibition of mycelial growth and of accumulation of oxalate. It is concluded that the EM pathway and the TCA cycle are involved in the formation of mycelial substance, CO2 and oxalate from gluconate. There is good correspondence between the rates of gluconate utilization and of oxalate accumulation which were observed immediately after the addition of gluconate and the in vitro activities of gluconokinase and oxaloacetate hydrolase, respectively, at this time.

Aspergillus niger↗
PubMed1997

[The effect of calcium gluconate on the acute and chronic toxicity of doxorubicin in mice].

The effect of calcium gluconate on the toxicity of the anthracycline antibiotic doxorubicin (DOX) in mice was studied. Calcium gluconate showed a significant protective action with respect to the DOX acute toxicity. When DOX was used in the lethal doses, up to the LD50s calcium gluconate protected all the mice from death. At the DOX LD100 or higher the antitoxic effect of calcium gluconate manifested itself in a lower death rate and/or in a higher lifespan of the animals (at least 2-fold). When DOX was used for the treatment course its chronic toxicity in the presence of calcium gluconate was 2 or more times lower by all the quantitative indices: the lifespan of the animals that died, the maximum and minimum total lethal doses of DOX, the latent period before the first mouse death, the overall duration of the DOX treatment course. The antitoxic effect of calcium gluconate also manifested itself in a lower DOX acute and chronic toxicity with respect to the gastrointestinal tract. Thus, calcium gluconate proved to be an effective DOX antitoxic modificator which provided the use of about 2 times higher single and courses doses of DOX in mice.

Animals↗
PubMed1991

Chlorhexidine gluconate and acetate in patch testing.

Patch testing to chlorhexidine is usually performed with chlorhexidine gluconate aq. We report the results of 297 patients, almost all with leg ulcers, concomitantly tested with chlorhexidine acetate 1% aq. and chlorhexidine gluconate 1% aq. 39 patients had positive reactions to one of these compounds or to both. 36 positive reactions to the acetate were found, in contrast to 18 reactions to the gluconate. The reactions were considered relevant in 22 of 39 patients, since these patients had developed an eczema in an area where a chlorhexidine compound was used, and discontinuing the chlorhexidine compound resulted in improvement of the condition. In 10 of these 22 patients, the diagnosis would have been missed if the gluconate only had been used for testing, while the acetate failed to diagnose 2 patients. In 109 patients without leg ulcers, inconclusive patch test readings (i.e., irritant reactions or weak positive reactions) were found in 17% with chlorhexidine acetate 1% aq., compared to 5% with chlorhexidine gluconate 1% aq., indicating a high degree of irritant potential of the acetate 1% aq. We consider that some positive reactions are lost if chlorhexidine gluconate 1% aq. only is used for patch testing, but that chlorhexidine acetate 1% aq., on the other hand, is an unacceptably strong irritant. We therefore suggest that further testing with chlorhexidine acetate 1 and 0.5% aq. should be performed, in parallel with chlorhexidine gluconate 1% aq., in order to establish appropriate test concentrations. We find that up to 13% of the leg ulcer patients in this study may be sensitized to chlorhexidine, and we recommend that the indications for the drug in leg ulcer patients should be reconsidered.

Chlorhexidine↗
PubMed2006

[Resorption and tolerance of the high doses of ferrous sulfate and ferrous gluconate in the patients on peritoneal dialysis].

BACKGROUND/AIM: Iron supplementation plays a crucial role in peritoneal dialysis (PD) patients. Oral iron substitution is more convenient than intravenous therapy in PD patients, but impaired absorption and adverse effects may be limiting factors for oral treatment. The aim of this study was to compare the absorption and side effects of high doses ferrous sulphate and ferrous gluconate in PD patients. METHODS: Blood samples were taken from 29 PD patients at baseline, as well as 2, 4 and 8 hours after oral intake of 4 ferrous sulphate tablets (containing 105 mg elemental iron per tablet). The test was repeated using 8 ferrous gluconate drinkable ampoules (containing 50 mg elemental iron per ampoule). RESULTS: The maximal increase in serum iron level during the test with iron sulphate was 113.51 +/- 103.37% versus the initial values of 183.87 +/- 37.38% during the ferrous gluconate test. The maximal values of serum iron after the intake of ferrous sulphate were 26.23 +/- 9.95 micromol/l versus 30.97 +/- 8.65 micromol/l after the intake of ferrous gluconate. There was a statistically significant difference between these two groups. Six patients showed an increase in serum iron of more than 300% after a high ferrous gluconate dose, while in 15 of the patients serum iron increased between 100% and 300%, and in 8 of the patients serum iron levels increased by less than 100%. Side effects occurred more frequently after the intake of ferrous sulphate than ferrous gluconate. CONCLUSION: High doses of oral iron were well absorbed and tolerated in PD patients. Ferrous gluconate was better absorbed and tolerated than ferrous sulphate, thus we recommend it for oral iron supplementation in PD patients.

Administration, Oral↗
PubMed1981

Gluconate calcium therapy and neonatal hypercalciuria.

Nephrolithiasis was present in a 2-month-old premature infant with bronchopulmonary dysplasia who had been receiving furosemide and intravenous (IV) gluconate calcium therapy. This infant was found to be hypercalciuric. Furosemide therapy is known to increase calcium excretion. In the present study, we examined sick infants who were receiving gluconate calcium without furosemide to evaluate the effect of gluconate calcium therapy on urinary calcium excretion. The sick infants receiving gluconate calcium had higher values of urinary calcium than did the well infants taking regular formula feedings. Moreover, the calciuria appeared to increase progressively with continued gluconate calcium therapy. It appears that prolonged use of either furosemide or IV gluconate calcium leads to hypercalciuria, which, in turn, may predispose the premature infant to nephrolithiasis.

Calcium↗
PubMed2005

Bioconversion of waste office paper to gluconic acid in a turbine blade reactor by the filamentous fungus Aspergillus niger.

Gluconic acid production was investigated using an enzymatic hydrolysate of waste office automation paper in a culture of Aspergillus niger. In repeated batch cultures using flasks, saccharified solution medium (SM) did not show any inhibitory effects on gluconic acid production compared to glucose medium (GM). The average gluconic acid yields were 92% (SM) and 80% (GM). In repeated batch cultures using SM in a turbine blade reactor (TBR), the gluconic acid yields were 60% (SM) and 67% (GM) with 80-100 g/l of gluconic acid. When pure oxygen was supplied the production rate increased to four times higher than when supplying air. Remarkable differences in the morphology of A. niger and dry cell weight between SM and GM were observed. The difference in morphology may have caused a reduction of oxygen transfer, resulting in a decrease in gluconic acid production rate in SM.

Air↗
PubMed1987

Calcium chloride versus calcium gluconate: comparison of ionization and cardiovascular effects in children and dogs.

A randomized prospective study in both children and dogs compared ionization of calcium chloride and calcium gluconate. Five conditioned dogs under halothane anesthesia received calcium chloride (4, 8, 12 mg/kg) and calcium gluconate (14, 28, 42 mg/kg) intravenously. Ten children scheduled for burn wound excision and grafting received both calcium chloride (2.5 mg/kg) and calcium gluconate (7.5 mg/kg) injected through a central venous cannula. Ionized calcium was measured at 0, 0.5, 1, 3, 5, and 10 min in the children, and 0, 0.5, 1, 2, 3, 4, 5, 10, 20, and 45 min in the dogs. The authors conclude that equal elemental calcium doses of calcium gluconate (10%) and calcium chloride (10%) (approximately 3:1), injected over the same period of time, are equivalent in their ability to raise [Ca++] during normocalcemic states in children and dogs; the changes in [Ca++] following calcium administration are short-lived (minutes); rapidity of ionization seems to exclude hepatic metabolism as an important factor in the dissociation of calcium gluconate; and equivalent rises in [Ca++] produced by calcium gluconate or calcium chloride resulted in equivalent cardiovascular effects. The authors feel that either form of calcium salt would be satisfactory if indicated during cardiopulmonary resuscitation or for the treatment of ionized hypocalcemia due to massive blood transfusion.

Adolescent↗
PubMed2005

Process optimization of continuous gluconic acid fermentation by isolated yeast-like strains of Aureobasidium pullulans.

This study was focused on the optimization of a new fermentation process for continuous gluconic acid production by the isolated yeast-like strain Aureobasidium pullulans DSM 7085 (isolate 70). Operational fermentation parameters were optimized in chemostat cultures, using a defined glucose medium. Different optima were found for growth and gluconic acid production for each set of operation parameters. Highest productivity was recorded at pH values between 6.5 and 7.0 and temperatures between 29 and 31 degrees C. A gluconic acid concentration higher than 230 g/L was continuously produced at residence times of 12 h. A steady state extracellular gluconic acid concentration of 234 g/L was measured at pH 6.5. 122% air saturation yielded the highest volumetric productivity and product concentration. The biomass-specific productivity increased steadily upon raising air saturation. An intracellular gluconic acid concentration of about 159 g/L (0.83 mol) was determined at 31 degrees C. This is to be compared with an extracellular concentration of 223 g/L (1.16 mol), which indicates the possible existence of an active transport system for gluconic acid secretion, or the presence of extracellular glucose oxidizing enzymes. The new process provides significant advantages over the traditional discontinuous fungi operations. The process control becomes easier, thus offering stable product quality and quantity.

Ascomycota↗
PubMed2006

Continuous gluconic acid production by the yeast-like Aureobasidium pullulans in a cascading operation of two bioreactors.

The application of a new developed process for the continuous production of gluconic acid using a cascade of two bioreactors in a continuous process is shown reaching the highest concentration of gluconic acid described in the literature for continuous culture fermentation. Very high gluconic acid concentrations of 272-308 g/l have been achieved under continuous cultivation of free-growing cells of Aureobasidium pullulans in the first bioreactor at residence times (RT) between 19.5 and 24 h with formation rates for the generic product between 12.7 and 13.9 g/(l h). Gluconic acid, 350-370 g/l, was continuously reached in the second bioreactor at a total RT of 30.8-37 h with R (j) of 9.2-12 g/(l h). The highest specific gluconic acid production (m (p)) of 3.6 g/(g h) was found in the first bioreactor at the lowest RT of 19.5 h. The highest selectivity of 93.6% was determined in the first bioreactor as well. Complete glucose consumption was obtained at 37 h total residence time in the second bioreactor. Gluconic acid, 433 g/l, was continuously produced in the second bioreactor at a total RT of 37 h.

Ascomycota↗
PubMed2006

Gluconic acid: an antifungal agent produced by Pseudomonas species in biological control of take-all.

Pseudomonas strain AN5 (Ps. str. AN5), a non-fluorescent Australian bacterial isolate, is an effective biological control (biocontrol) agent of the take-all disease of wheat caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Ps. str. AN5 controls Ggt by producing an antifungal compound which was purified by thin layer and column chromatography, and identified by NMR and mass spectroscopic analysis to be d-gluconic acid. Commercially bought pure gluconic acid strongly inhibited Ggt. Two different transposon mutants of Ps. str. AN5 which had lost take-all biocontrol did not produce d-gluconic acid. Gluconic acid production was restored, along with take-all biocontrol, when one of these transposon mutants was complemented with the corresponding open reading frame from wild-type genomic DNA. Gluconic acid was detected in the rhizosphere of wheat roots treated with the wild-type Ps. str. AN5, but not in untreated wheat or wheat treated with a transposon mutant strain which had lost biocontrol. The antifungal compounds phenazine-1-carboxylic acid and 2,4-diacetylphloroglucinol, produced by other Pseudomonads and previously shown to be effective in suppressing the take-all disease, were not detected in Ps. str. AN5 extracts. These results suggest that d-gluconic acid is the most significant antifungal agent produced by Ps. str. AN5 in biocontrol of take-all on wheat roots.

Antifungal Agents↗
PubMed2002

Conformational changes during the catalytic cycle of gluconate kinase as revealed by X-ray crystallography.

The crystal structure of gluconate kinase from Escherichia coli has been determined to 2.0 A resolution by X-ray crystallography. The three-dimensional structure was solved by multi-wavelength anomalous dispersion, using a crystal of selenomethionine-substituted enzyme. Gluconate kinase is an alpha/beta structure consisting of a twisted parallel beta-sheet surrounded by alpha-helices with overall topology similar to nucleoside monophosphate (NMP) kinases, such as adenylate kinase. In order to identify residues involved in substrate binding and catalysis, structures of binary complexes with ATP, the ATP analogue adenosine 5'-(beta,gamma-methylene) triphosphate and the product, gluconate-6-phosphate have been determined. Significant conformational changes are induced upon binding of ATP to the enzyme. The largest changes involve a hinge-bending motion of the NMP(bind) part and a motion of the LID with adjacent helices, which opens the cavity to the second substrate, gluconate. Opening of the active site cleft upon ATP binding is the opposite of what has been observed in the NMP kinase family so far, which usually close their active site to prevent fortuitous hydrolysis of ATP. The conformational change positions the side-chain of Arg120 to stack with the purine ring of ATP and the side-chain of Arg124 is shifted to interact with the alpha-phosphate in ATP, at the same time protecting ATP from solvent water. The beta and gamma-phosphate groups of ATP bind in the predicted P-loop. A conserved lysine side-chain interacts with the gamma-phosphate group, and might promote phosphoryl transfer. Gluconate-6-phosphate binds with its phosphate group in a similar position as the gamma-phosphate of ATP, consistent with inline phosphoryl transfer. The gluconate binding-pocket in GntK is located in a different position than the nucleoside binding-site usually found in NMP kinases.

Adenosine Triphosphate↗
PubMed2005

[Assessment of the photosensitization potential of zinc gluconate].

INTRODUCTION: Tolerance and clinical efficacy of zinc gluconate are well documented, however, no study have evaluated its photosensitizing potential. It is well known that many treatments of acne are photosensitizing. Evaluation of the photosensitizing potential of zinc gluconate was the aim of this study. PATIENTS AND METHOD: Two open, monocentric studies were carried out with acneic volunteers. The methodology used in this study was an adaptation from that existing for the evaluation of the photosensitizing potential of topical products. In the study of phototoxic potential, volunteers were exposed to 20 J/cm2 UVA and to 0.75 times MED, before and after administration of zinc gluconate. Clinical and colorimetric evaluations of reactions were then carried out 1, 24, 48 and 72 hrs after exposure. In the photoallergic potential study, during the first week of the induction phase, the volunteers were exposed to 2 times MED, and to 3 times the MED during the second and third week. Then during the challenge phase, they were exposed t o4 J/cm2 UVA and to 0.75 times MED. Zinc gluconate was administered throughout the study. Clinical and colorimetric evaluations of reactions were carried out 24, 48 and 72 hrs after exposure, only during the challenge phase. RESULTS: The majority of clinical scores measured on scales at 6 and 5 levels were equal to 0 and 0.5 (evaluation of the phototoxic potential) or all equal to 0 (evaluation of the photoallergic potential). Thus zinc gluconate did not induce phototoxic or photosensitive reactions, whatever the ultraviolet type used. DISCUSSION: Since zinc gluconate does not induce any photosensitive reaction, it could be prescribed during periods of exposure to sun.

Acne Vulgaris↗
PubMed2005

Potential application of a glucose-transport-deficient mutant of Schizosaccharomyces pombe for removing gluconic acid from grape must.

Musts from rotten grapes typically contain high levels of gluconic acid, which can raise severe problems in winemaking processes. In this work, the ability of the glucose-transport-deficient mutant YGS-5 of Schizosaccharomyces pombe to completely or partly remove gluconic acid from a synthetic glucose-containing medium and the potential use of this yeast strain for the same purpose in musts and wines were examined. Surprisingly, the S. pombe YGS-5 strain successfully removed 93% of the initial gluconic acid (2.5 gL(-1)) and 80% of the initial malic acid (1.0 gL(-1)) within 30 h after inoculation. Also, the yeast strain produced no volatile compounds other than those obtained in fermentations conducted with the wine yeast Saccharomyces cerevisiae. S. pombe YGS-5 could thus be used to remove gluconic acid present in musts from rotten grapes. On the basis of these results, various ways of using S. pombe YGS-5 to treat musts containing gluconic acid in order to solve the problems due to the high gluconic acid concentrations in botrytized grape must are proposed.

Fermentation↗