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DNA microarray analyses of the long-term adaptive response of Escherichia coli to acetate and propionate.

In its natural environment, Escherichia coli is exposed to short-chain fatty acids, such as acetic acid or propionic acid, which can be utilized as carbon sources but which inhibit growth at higher concentrations. DNA microarray experiments revealed expression changes during exponential growth on complex medium due to the presence of sodium acetate or sodium propionate at a neutral external pH. The adaptive responses to acetate and propionate were similar and involved genes in three categories. First, the RNA levels for chemotaxis and flagellum genes increased. Accordingly, the expression of chromosomal fliC'-'lacZ and flhDC'-'lacZ fusions and swimming motility increased after adaptation to acetate or propionate. Second, the expression of many genes that are involved in the uptake and utilization of carbon sources decreased, indicating some kind of catabolite repression by acetate and propionate. Third, the expression of some genes of the general stress response increased, but the increases were more pronounced after short-term exposure for this response than for the adaptive response. Adaptation to propionate but not to acetate involved increased expression of threonine and isoleucine biosynthetic genes. The gene expression changes after adaptation to acetate or propionate were not caused solely by uncoupling or osmotic effects but represented specific characteristics of the long-term response of E. coli to either compound.

Acetates↗

Carbon-13 nuclear magnetic resonance study of metabolism of propionate by Escherichia coli.

We have evaluated the use of [1,2-13C2]propionate for the analysis of propionic acid metabolism, based on the ability to distinguish between the methylcitrate and methylmalonate pathways. Studies using propionate-adapted Escherichia coli MG1655 cells were performed. Preservation of the 13C-13C-12C carbon skeleton in labeled alanine and alanine-containing peptides involved in cell wall recycling is indicative of the direct formation of pyruvate from propionate via the methylcitrate cycle, the enzymes of which have recently been demonstrated in E. coli. Additionally, formation of 13C-labeled formate from pyruvate by the action of pyruvate-formate lyase is also consistent with the labeling of pyruvate C-1. Carboxylation of the labeled pyruvate leads to formation of [1,2-13C2]oxaloacetate and to multiply labeled glutamate and succinate isotopomers, also consistent with the flux through the methylcitrate pathway, followed by the tricarboxylic acid (TCA) cycle. Additional labeling of TCA intermediates arises due to the formation of [1-13C]acetyl coenzyme A from the labeled pyruvate, formed via pyruvate-formate lyase. Labeling patterns in trehalose and glycine are also interpreted in terms of the above pathways. The information derived from the [1, 2-13C2]propionate label is contrasted with information which can be derived from singly or triply labeled propionate and shown to be more useful for distinguishing the different propionate utilization pathways via nuclear magnetic resonance analysis.

Acyl Coenzyme A↗

Identification of two prpDBC gene clusters in Corynebacterium glutamicum and their involvement in propionate degradation via the 2-methylcitrate cycle.

Genome sequencing revealed that the Corynebacterium glutamicum genome contained, besides gltA, two additional citrate synthase homologous genes (prpC) located in two different prpDBC gene clusters, which were designated prpD1B1C1 and prpD2B2C2. The coding regions of the two gene clusters as well as the predicted gene products showed sequence identities of about 70 to 80%. Significant sequence similarities were found also to the prpBCDE operons of Escherichia coli and Salmonella enterica, which are known to encode enzymes of the propionate-degrading 2-methylcitrate pathway. Homologous and heterologous overexpression of the C. glutamicum prpC1 and prpC2 genes revealed that their gene products were active as citrate synthases and 2-methylcitrate synthases. Growth tests showed that C. glutamicum used propionate as a single or partial carbon source, although the beginning of the exponential growth phase was strongly delayed by propionate for up to 7 days. Compared to growth on acetate, the specific 2-methylcitrate synthase activity increased about 50-fold when propionate was provided as the sole carbon source, suggesting that in C. glutamicum the oxidation of propionate to pyruvate occurred via the 2-methylcitrate pathway. Additionally, two-dimensional gel electrophoresis experiments combined with mass spectrometry showed strong induction of the expression of the C. glutamicum prpD2B2C2 genes by propionate as an additional carbon source. Mutational analyses revealed that only the prpD2B2C2 genes were essential for the growth of C. glutamicum on propionate as a sole carbon source, while the function of the prpD1B1C1 genes remains obscure.

Base Sequence↗

Transport of propionate by human ileal brush-border membrane vesicles.

Human ileal brush-border membrane vesicles were employed to study the mechanisms of short-chain fatty acid (propionate) absorption especially to determine the effects of intravesicular HCO3- and the component of nonionic diffusion. Preloading the vesicles with HCO3- resulted in up to 20-fold "overshoots" of transport, and this effect was not seen with other intravesicular anions. This transport process was very fast (peak uptake 6 s) and was not due to intravesicular buffering by HCO3-. Radiolabeled propionate transport demonstrated transstimulation when the vesicles were preloaded with unlabeled propionate. An inward H+ gradient led to stimulation of propionate transport much smaller than in the presence of trans-HCO3-, whereas an inward Na+ gradient had no effect. Propionate transport was attenuated by the anion exchange inhibitors SITS and DIDS. Under HCO3- gradient conditions, propionate transport exhibited saturation kinetics with an apparent Km of 21 +/- 3 mM and a Vmax of 50 +/- 3 nmol.mg protein-1.3 s-1. Propionate transport was inhibited up to 40% by 2-5 carbon short-chain fatty acids (10 mM) but not by other organic anions. Short-chain fatty acid transport in the human ileum is Na+ independent and occurs mostly via a specific anion exchange mechanism with HCO3-. Our results also demonstrate a small component of nonionic diffusion of the protonated fatty acid (or anion exchange for OH-).

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Contrasting effects of propionate and propionyl-L-carnitine on energy-linked processes in ischemic hearts.

Propionyl-L-carnitine, unlike L-carnitine, is known to improve myocardial function and metabolism altered during the course of ischemia-reperfusion. In this study, the effect of propionyl-L-carnitine has been compared with that of propionate and carnitine on the performance of rat hearts perfused with a glucose-containing medium either under normoxia, ischemia, or postischemic reperfusion. In the postischemic phase, contractile parameters were partially restored both in the control and in the propionate plus carnitine-treated hearts, were markedly impaired by propionate, and were fully recovered by propionyl-L-carnitine. In addition, propionyl-L-carnitine, but not propionate, reduced the functional decay of mitochondria prepared from the ischemic hearts. Even in normoxic conditions propionate, unlike propionyl-L-carnitine, caused a drastic reduction of free CoA and L-carnitine. The concomitant increase in lactate production and decrease in ATP content might be explained by the inhibition of pyruvate dehydrogenase caused by the accumulation of propionyl-CoA. Indeed, when pyruvate was the only oxidizable substrate, propionate induced a gradual decrease in developed pressure, which was largely prevented by L-carnitine. The protective effect of propionyl-L-carnitine may be a consequence of the anaplerotic utilization of propionate in the presence of an optimal amount of ATP and free L-carnitine.

Animals↗

Pancytopenia in propionic acidemia: hematologic evaluation and studies of hematopoiesis in vitro.

This study investigated the hematologic abnormalities of an infant with propionic acidemia and reversible pancytopenia. Light and electron microscopy of her bone marrow revealed severely disturbed cellular morphology with trilineage dysmyelopoiesis, hemophagocytosis, and numerous multinucleated histiocytes and megakaryocytes. The effects of her serum and of organic acids associated with propionic acidemia were studied on hematopoiesis in vitro. Mouse erythroid (CFU-E) and granulocyte-monocyte colonies (CFU-GM) were assayed by fibrin clot technique; human CFU-GM were grown in agar culture. The infant's serum reduced mouse CFU-E and CFU-GM by 43 and 32%, respectively, compared with normal human sera, but had no effect on human CFU-GM in our culture system. Buffered propionic acid caused concentration-dependent inhibition of mouse CFU-E and human CFU-GM over a range reported in sera of acutely ill infants with propionic acidemia. Neither cell viability nor subsequent colony formation was diminished by preincubation of bone marrow cells with propionic acid for 48 h. The three other organic acids studied, tiglic acid, 3-OH propionate, and glycine, did not inhibit growth of mouse CFU-E, CFU-GM, or human CFU-GM, and glycine significantly enhanced formation of the latter. Evaluation of the infant's hematologic abnormalities suggests that inhibition of bone marrow proliferation and maturation and, perhaps, shortened red blood cell survival were responsible for her pancytopenia. The studies performed in vitro implicate propionic acid in this hematopoietic dysfunction.

Amino Acid Metabolism, Inborn Errors↗

Using a feed-grade zinc propionate to achieve molt induction in laying hens and retain postmolt egg production and quality.

A commercial-feed-grade form of zinc propionate was examined as a potential feed amendment at a concentration of 1% zinc to induce molt in 90-wk-old hens. Dietary treatments consisted of 4 treatment groups of 28 birds each randomly assigned to either (1) molted conventionally by feed withdrawal, (2) 1% zinc as Zn acetate, (3) 1% zinc as Zn propionate, or (4) nonmolted control for 9 d. Ovary weights of hens fed Zn acetate or Zn propionate were not significantly different from each other, but hens fed Zn acetate or Zn propionate were significantly (p<0.05) lighter than the ovary weight of nonmolted control hens. Zinc concentrations in the kidney and liver were significantly (p<0.05) increased in both Zn acetate- and Zn propionate-molted hens when compared to either nonmolted control-fed hens or feed-withdrawal molted hens. Over the entire 3-mo postmolt period, there were no significant differences in interior or exterior egg qualities among the four treatments. Egg production of hens fed Zn acetate was significantly lower than feed-withdrawal hens, Zn propionate-fed hens, or nonmolted control hens (p<0.05). The data of the current study demonstrated that feeding a feed grade of Zn propionate (1% Zn)-supplemented diet can induce molt and retain postmolt egg quality and production comparable to hens molted by feed withdrawal.

Animal Feed↗

Propionic acid disappearance from the foregut and small intestine of the beef steer.

Studies were conducted to determine the disappearance of propionic acid from the rumen and across the small intestine. Four crossbred beef steers, fitted with ruminal, duodenal and ileal cannulas, were given pulsed-continuous infusions of a nonabsorbable liquid phase marker (CrEDTA) and buffered propionic acid to achieve ruminal propionate productions (RPP) of 142 (basal), 567 and 997 mmol/h. Liquid flows from the rumen, and at the duodenum and ileum (2.75, 3.57 and .92 liters/h, respectively) were not affected by RPP (P greater than .23), although significant differences existed between sites (P = .001), reflecting a net addition of liquid between the rumen and duodenum and a net removal of liquid within the small intestine. Propionate disappearance from the rumen was 40 to 57% irrespective of RPP, with the complement passing from the rumen. Of the propionate that exited the rumen, 93 to 97% disappeared prior to entering the duodenum. Overall removal of RPP prior to the duodenum was 97 to 99% irrespective of RPP. Passage of propionic acid at the duodenum and at the ileum did not differ (P greater than .76) across all RPP, and the overall mean passages did not differ from zero (P greater than .34); similar results were observed for other VFA. These observations indicate that essentially all RPP is removed prior to entering the small intestine. Additionally, net propionate disappearance across the small intestine was undetectable and likely biologically irrelevant. Therefore, VFA absorption does not appear to be a limiting biological process in the beef steer.

Animals↗

Effects of isoenergetic infusions of propionate and glucose on portal-drained visceral nutrient flux and concentrations of hormones in lambs maintained by total intragastric infusion.

Three lambs were used in a repeated Latin square design to determine the influence of isoenergetic infusions of propionate or glucose on portal-drained visceral flux (PDV) of nutrients and concentrations of insulin, glucagon, growth hormone and prolactin. Lambs were fitted with appropriate catheters for blood sampling and maintained on total intragastric infusion of nutrients. Basal VFA, casein, mineral and vitamin infusions (isocaloric and isonitrogenous) were supplemented with an additional 22 +/- .5 kcal/h from propionate, glucose or a combination of propionate plus glucose. Ruminal fluid proportion and arterial blood concentration and PDV flux of propionate increased (P less than .10) by 17 mol/100 mol, .02 mM and 40 mmol/h, respectively, with infusion of an additional 61 mmol/h of propionate. Regression equations predicted that, on a net basis, 67% of ruminally infused propionate and 43% of abomasally infused glucose appeared in portal blood. Arterial L-lactate, beta-hydroxybutyrate and acetate concentrations, and beta-hydroxybutyrate flux were increased (P less than .10) by .34 mM, .20 mM, .50 mM and 4.2 mmol/h, respectively, with infusion of 33 mmol/h of added glucose. Net utilization of glucose by the PDV was approximately 4.4 mmol/h when no glucose was infused. Increased infusion of propionate resulted in a 22.2-micrograms/h increase in PDV flux of insulin (P less than .08) but had no effect on arterial insulin, glucagon and prolactin concentrations (P greater than .10). Arterial growth hormone increased by 3.8 ng/ml with increasing glucose infusion (P less than .08).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Plasma insulin and glucagon responses to propionate infusion into femoral and mesenteric veins in sheep.

Propionate (1, 2, 4, 8, 16, 32, and 64 mumol.kg BW-1.min-1 for 30 min) was infused into the femoral and mesenteric veins of adult sheep to investigate the physiological significance of propionate in regulating plasma insulin and glucagon concentrations. The increments in arterial blood propionate concentrations during propionate infusion increased (P < .001) with increasing infusion rates for both infusion sites, and they were smaller (P < .001) for the mesenteric vein infusion than for the femoral vein infusion. Plasma insulin concentrations during propionate infusion increased (P < .10) from preinfusion values with infusion rates of > or = 8 mumol.kg BW-1.min-1 for both infusion sites. The response areas of plasma insulin concentration above basal tended to be smaller (P < .112) for the mesenteric vein infusion than for the femoral vein infusion. Plasma glucagon concentrations during propionate infusion increased (P < .05) from preinfusion values with infusion rates of > or = 8 and 64 mumol.kg BW-1.min-1 for the femoral and mesenteric vein infusions, respectively. The response areas of plasma glucagon concentration above basal were smaller (P < .011) for the mesenteric vein infusion than for the femoral vein infusion. We conclude that in sheep propionate absorbed from the alimentary tract has a physiological role in regulating circulating concentrations of insulin and glucagon.

Animals↗

Preference for flavored wheat straw by lambs conditioned with intraruminal administrations of sodium propionate.

We hypothesized that volatile fatty acids are feedback signals that condition food preferences in ruminants, and we tested two predictions based on this hypothesis: 1) low doses of propionate condition preferences for low-quality foods (Exp. 1 and 2) preferences are not caused by osmotic load (Exp. 2). In Exp. 1, lambs were offered chopped wheat straw flavored with either oregano or onion on odd days, whereas on even days flavors were switched and lambs received capsules containing sodium propionate. During four 8-d conditioning periods, the amounts of propionate delivered ranged from .7 to 1.4% of the daily DE intake (Period 1) or were fixed at .7% (Period 2) and 1% of the daily DE intake (Periods 3 and 4). After each 8-d conditioning period, lambs were offered oregano- and onion-flavored straw. Conditioning was then suspended and lambs were offered onion- and oregano-flavored straw at weekly intervals for 1 mo (extinction). Lambs preferred the flavor paired with propionate during conditioning (P < .001) and extinction (P < .07). During Exp. 2, a different group of lambs was conditioned as in Exp. 1, but sodium chloride was delivered at osmotic loads equivalent to those when propionate supplied .7% and 1% of the daily DE intake. Lambs strongly avoided the flavor paired with sodium chloride (P < .001). Thus, lambs acquired preferences for straw conditioned with doses of propionate typically considered ineffective in the regulation of food intake, and osmolalities generated by propionate did not cause, but probably attenuated, food preferences.

Animals↗

Counteractive effects of propionate or 1,2-propanediol against hypoglycemia and ketonemia of tributyrin-treated cows.

We administered tributyrin (500 ml), tributyrin (500 ml) plus magnesium propionate (400 g), tributyrin (500 ml) plus sodium propionate (400 g), or tributyrin (500 ml) plus 1,2-propanediol (400 ml) as a single dose into rumens of lactating cows and then measured in blood the plasma concentrations of glucose, acetoacetate, 3-hydroxybutyrate, free fatty acids, and insulin as a function of time. Tributyrin administration caused hypoglycemia and hyperketonemia similar to the ketotic condition in less than 3 h and was a negative correlation of --.88 between glucose and ketone concentrations in blood plasma. Administration of either magnesium propionate, sodium propionate, or 1,2-propanediol could counteract the hypoglycemia and hyperketonemia induced by tributyrin administration without significantly changing the insulin response. Of the two propionate compounds, magnesium propionate was more effective than sodium propionate for alleviating hypoglycemia and hyperketonemia.

3-Hydroxybutyric Acid↗

Effects of propionate and methylmalonate on conversions of acetate, butyrate, and D(-)-3-hydroxybutyrate to fatty acids and carbon dioxide by mammary tissue slices of goats.

Incorporations of [1-carbon-14] acetate, [1-carbon-14] propionate, n-[1-carbon-14] butyrate, and D(-)-3-hydroxy[3-carbon-14] butyrate into individual milk fatty acids and their conversion to carbon dioxide were studied in vitro with caprine mammary tissue slices in the presence and absence of propionate and methylmalonate. Neither propionate nor methylmalonate affected incorporation of these substances into fatty acids. In a decreasing order butyrate, acetate, propionate, and D(-)-3-hydroxybutyrate were converted to carbon dioxide. Acetate had the highest incorporation rate into fatty acids followed by D(-)-3-hydroxybutyrate, butyrate, and propionate. Labeled propionate was incorporated mainly into odd-numbered fatty acids. Results do not support the theory that either propionate or its metabolite, methylmalonate, inhibit de novo synthesis of fatty acids in the mammary gland in relation to the etiology of low milk fat syndrome.

3-Hydroxybutyric Acid↗

Effect of mesenteric vein infusion of propionate on splanchnic metabolism in primiparous Holstein cows.

Our objective was to assess the effects of increased propionate supply on gut and liver function in lactating cows. Four multicatheterized, primiparous cows (30.4 +/- .5 kg/d of milk) were fed for ad libitum intake a diet of 50% alfalfa hay and 50% concentrate (20.6 +/- 1.9 kg/d of DM, 226 +/- 21 MJ/d of metabolizable energy, and 611 +/- 56 g/d of N). Each cow received intramesenteric infusions of NaCl (control) or Na-propionate (150 mmol/h of a 2.5 M solution) in a reversal design. After 72 h of infusion, blood flow (by indicator dilution) and net flux (venoarterial differences multiplied by blood flow) were measured across portal-drained viscera and the liver. Energy supply from feed consumed and from infusion was similar between treatments. Energy that was excreted as milk decreased with propionate infusion. Propionate infusion increased arterial concentration of propionate; decreased absorption of acetate, butyrate, and valerate; and decreased hepatic removal of L-lactate, butyrate, valerate, NEFA, and oxygen. Propionate infusion decreased splanchnic release of glucose and increased splanchnic release of acetate and alanine. Net flux of urea, BHBA, insulin, or glucagon was unaffected by treatments. Our data show a link between a greater proportion of energy supplied as propionate and decreased energy excreted as milk. This response was associated with decreased net removal of glucogenic and ketogenic substrates by the liver and increased supply of acetate for use by peripheral tissues.

Analysis of Variance↗

The importance of pH in the regulation of ruminal acetate to propionate ratio and methane production in vitro.

Grain feeding often causes a decrease in ruminal pH, and experiments were conducted to define the role of pH in regulating the acetate to propionate ratio and production of CH4. Cows that were fed 90% concentrate had lower ruminal pH values (6.22 vs. 6.86), higher VFA concentrations (85 vs. 68 mM), and lower acetate to propionate ratios (2.24 vs. 4.12) than did cows that were fed forage only. When mixed ruminal bacteria from cows that were fed 90% concentrate or 100% forage were incubated (48 h) with hay (10 g/L) or cracked corn (5 g/L) in a medium containing bicarbonate (38 mM) and tricarballylate (50 mM), the final pH values were less than 0.3 units lower than the initial pH. At final pH values less than 5.7, hay fermentation was inhibited, the acetate to propionate ratio and CH4 production declined more than twofold, and the inoculum source was without effect. Small amounts of H2 were detected at pH values less than 5.5. Total VFA production from cracked corn decreased when pH declined, but only if the inoculum was obtained from cows that were fed 90% concentrate. The acetate to propionate ratio of cracked corn incubations declined from 1.2 to 0.6 when final pH was decreased from 6.5 to 5.3, and CH4, as a percentage of total VFA production, also decreased. At pH values less than 5.3, the acetate to propionate ratio of cracked corn increased more than fourfold, and large amounts of H2 could be detected. Over the final pH range of 6.5 to 5.3, CH4 production was highly correlated with acetate to propionate ratio, which was dependent on pH and substrate (CH4 = 0.02 + 0.05 pH; r2 = 0.80). Calculations based on the differences between pH 6.5 and 5.8 indicated that as much as 25% of the decrease in acetate to propionate ratio could be explained by the effect of pH alone.

Acetates↗

Propionate inhibits hepatocyte lipid synthesis.

Oat bran lowers serum cholesterol in animals and humans. Propionate, a short-chain fatty acid produced by colonic bacterial fermentation of soluble fiber, is a potential mediator of this action. We tested the effect of propionate on hepatocyte lipid synthesis in rats using [1-14C]acetate, 3H2O, and [2-14C]mevalonate as precursors. Propionate produced a statistically significant inhibition of cholesterol biosynthesis from [1-14C]acetate at a concentration of 1.0 mM and from 3H2O and [2-14C]mevalonate at concentrations of 2.5 mM. Propionate also produced a significant inhibition of fatty acid biosynthesis at concentrations of 2.5 mM using [1-14C]acetate as a precursor. The demonstration of propionate-mediated inhibition of cholesterol and fatty acid biosynthesis at these concentrations suggests that propionate may inhibit cholesterol and fatty acid biosynthesis in vivo and may mediate in part the hypolipidemic effects of soluble dietary fiber. Further studies are needed to clarify this action of propionate and to establish the exact mechanisms by which the inhibition occurs.

Animals↗

[Role of propionic acid, yeasts and ethyl alcohol in regulating the activity of H-factor in Drosophila simulans].

Effects of yeast, propionic acid and ethanol on the activity of H-factor, which sharply increases the frequency of somatic recombination in X-chromosomes of dorsal prothoracal disc cells in Drosophila simulans are studied. The frequency of yellow and singed mosaic spots in heterozygous yw ++/++sn1vHD. melanogaster females, inherited H-factor from the father (the stock sn1v) is estimated. The results of the varience analysis have shown that yeast and propionic acid regulate the activity of H-factor in cells of dorsal prothoracal disc, the interaction of yeast and propionic acid being also observed. Yeasts (or some unknown product of their metabolism) are the activator of H-factor; thus, when larvae eat much yeast, the frequency of yellow and singed mosaic spots in humeral region becomes high. A decrease of mosaic spot frequency under the increase of propionic acid content in nutrition medium is a result of the inhibitory effect of propionic acid on the yeast growth, but not of the direct repression of H-factor activity. So, propionic acid may be considered as a regulator of the second order. Ethanol does not activate H-factor. Changes in the content of yeast and propionic acid in nutrition medium do not affect the frequency of yellow and singed mosaic spots in other regions of D. simulans body, except humeral.

Animals↗

Effect of propionate on lipogenesis in adipose tissue.

The metabolism of propionate in adipose tissue and its effect on lipogenesis was investigated. Fasting induced changes in propionate metabolism of adipose tissue, drastically reducing higher fatty acid synthesis and increasing glyceride-glyerol formation from low concentrations of propionate (0.25 mM). Propionate also promoted lipogenesis from acetate-1-(14)C in tissues of fasted rats, while it inhibited lipogenesis and CO(2) formation from acetate in the fed animal. Treatment with actinomycin D or ethionine abolished both the increased glyceride-glycerol formation from propionate and the promoting effect on lipogenesis from acetate. Synthesis of long-chain fatty acids from propionate-1-(14)C was increased by actinomycin treatment. The change in propionate metabolism induced by fasting is, however, not entirely due to its conversion to glyceride-glycerol, since the latter was almost completely blocked by malonate while part of the promoting effect on fatty acid synthesis persisted.

Acetates↗