Utilization of short chain monocarboxylic acids in an effluent of a petrochemical industry by Acinetobacter calcoaceticus

The aqueous effluent generated by the Fischer--Tropsch process, containing a total of 13 g/L C(2)-C(5) monocarboxylic acids, was investigated as a potential substrate for the production of single-cell protein (SCP). A bacterial isolate, Acinetobacter calcoaceticus, could utilize all the acids in the effluent simultaneously in chemostat cultures, and no residual acids were detected in the culture below a dilution rate of 0.78 h(-1). The critical dilution rate was 1.04 h(-1). The maintenance energy requirement of the cells growing on the monocarboxylic acid mixture was considerably lower than that of cells growing on acetate as the sole carbon source. Enrichment of the effluent with ethanol to increase the biomass concentration was successful and still allowed the simultaneous and efficient utilization of all the carbon sources, but resulted in a decrease of the critical dilution rate by ca. 20%.     

Molecular Distribution of Monocarboxylic Acids in Asuka Carbonaceous Chondrites from Antarctica

Molecular distribution of low-molecular-weight monocarboxylic acids was studied in three CM2 Asuka carbonaceous chondrites (A-881280, A-881334 and A-881458), which were recovered from Antarctica by the 29th Japanese Antarctic Research Expedition in 1988. GC and GC/MS analyses identified more than 30 monocarboxylic acids in A-881458, including aliphatic and aromatic acids with various structural isomers. Isomeric phenolic compounds were also identified. The aliphatic carboxylic acids have straight-chain structures having 2 to 12 carbon atoms (C2 to C12), and branched-chain structures (C4 to C9). The quantities of straight-chain acids decrease logarithmically with increasing carbon number. At the same carbon number, a straight-chain isomer is always predominant compared to branched-chain isomers. All of the 14 possible C4, C5 and C6 aliphatic monocarboxylic acids (not including optical isomers) have been identified, although all the isomers were not reported in Murchison and Y-791198 meteorites. Of the 17 possible isomeric C7 acids, at least 14 isomers were tentatively identified by mass spectra (EI and CI mode). At C8 or above, peaks of branched-chain isomers become obscure, probably due to the large number of isomers and small concentrations. Branched-chain monocarboxylic acids over C6 have never been reported in Murchison. Although occurrence of aliphatic acids are similar between A-881458 and Murchison at C4, C5 and C6 acids, a major difference is that A-881458 as well as Y-791198 have straight- chain predominance among isomers in contrast to Murchison being branched-chain predominant. In the case of isomeric aromatic compounds such as toluic acids and cresols, m-toluic acid and p-cresol are more abundant among their isomers, respectively. The molecular distribution may not reflect thermodynamic equilibrium but rather a kinetically controlled process for their formation mechanism. The other two CM2 chondrites (A-881280 and A-881334) were depleted in carboxylic acids in spite of similar carbon contents. The depletion is not due to weathering on ice, because the degrees of weathering are small and similar among the three chondrites. Probably those latter two chondrites may have been subjected to aqueous alteration or metamorphism on their meteorite parent bodies.     

Chemiluminescence of the peroxomonosulphate-cobalt(II)-aliphatic monocarboxylic acids system.

A weak chemiluminescence (CL) emission was observed due to the production of singlet oxygen ((1)O(2)) during the decomposition of peroxomonosulphate (HSO(5)(-)) catalysed by cobalt(II). Low molecular mass aliphatic monocarboxylic acids, such as formic, acetic, propionic, butyric and valeric acids, influenced the CL emission, and the reaction of aliphatic monocarboxylic acids with HSO(5)(-)/Co(2+) solution was further investigated using a flow injection analysis (FIA) CL method. The results indicated that the CL intensities of aliphatic monocarboxylic acids were improved with increase in the carbon chain length in the potassium peroxomonosulphate-cobalt(II) sulphate system. Generation of singlet oxygen was confirmed by the fact that the CL emission of aliphatic monocarboxylic acids with the HSO(5)(-)/Co(2+) solution was quenched by NaN(3), and from the CL spectrum of the reaction system. Additionally, a possible mechanism of aliphatic monocarboxylic acids CL emission enhancement was proposed.     

Low-molecular-weight aliphatic carboxylic acids in soil solutions under different vegetations determined by capillary zone electrophoresis

Concentrations of low-molecular-weight aliphatic carboxylic acids in soil solution were determined by a newly developed capillary zone electrophoresis method. Soil solution samples were collected by centrifugation of soil from the A horizon of a Danish, homogeneous, nutrient-rich Hapludalf in adjacent forested and arable plots. The forested plots of 0.5 ha were 33-year old stands of beech (Fagus sylvatica L.), oak (Quercus robur L.), grand fir (Abies grandis Lindl.), and Norway spruce (Picea abies (L.) Karst.), while sugar beet (Beta vulgaris L.) and winter wheat (Triticum aestivum L.) were the agricultural crops this year. High variability in soil solution concentrations of metal cations (Al, Ca, K, Mg, Na), monocarboxylic acids (formic, acetic, lactic, and valeric acids), and di- and tricarboxylic acids (oxalic, malic, succinic, and citric acids) were found within each plot. Despite this short-range within-plot variability, higher concentrations of di- and tricarboxylic acids were found in the forested soils than in the arable soils. The vegetation seemed to favour some monocarboxylic acids, but the total monocarboxylic acid concentrations showed little relation to the vegetation. Probably due to much less soil water in the Norway spruce plot, the low-molecular-weight aliphatic carboxylic acid concentrations in the samples from that plot were much higher than those found in samples from the other plots. Carbon in low-molecular-weight aliphatic carboxylic acids only accounts for a few percent of dissolved organic carbon, and no general relation was found between carbon in low-molecular-weight aliphatic carboxylic acids and dissolved organic carbon, although the correlation between carbon in di- and tricarboxylic acids and dissolved organic carbon was significant. This revised version was published online in June 2006 with corrections to the Cover Date.     

Long-chain α,ω-dioic acids as inducers of cyclosporin A-insensitive nonspecific permeability of the inner membrane of liver mitochondria loaded with calcium or strontium ions

Long-chain saturated monocarboxylic fatty acids can induce nonspecific permeability of the inner membrane (open pores) of liver mitochondria loaded with Ca²⁺ or Sr²⁺ by the mechanism insensitive to cyclosporin A. In this work we investigated the effect of their metabolites — α,ω-dioic (dicarboxylic) acids — as potential inducers of pore opening by a similar mechanism. It was established that the addition of α,ω-hexadecanedioic acid (HDA) at a concentration of 10–30 μM to liver mitochondria loaded with Ca²⁺ or Sr²⁺ leads to swelling of the organelles and release of these ions from the matrix. The maximum effect of HDA is observed at 50 μM Ca²⁺ concentration. Cyclosporin A at a concentration of 1 μM, previously added to the mitochondria, did not inhibit the observed processes. The calcium uniporter inhibitor ruthenium red, which blocks influx of Ca²⁺ and Sr²⁺ to the matrix of mitochondria, prevented HDA-induced swelling. The effect of HDA as inducer of swelling of mitochondria was compared with similar effects of α,ω-tetradecanedioic and α,ω-dodecanedioic acids whose acyl chains are two and four carbon atoms shorter than HDA, respectively. It was found that the efficiency of these α,ω-dioic acids decreases with reducing number of carbon atoms in their acyl chains. It was concluded that in the presence of Ca²⁺ or Sr²⁺ long-chain saturated α,ω-dioic acids can induce a cyclosporin A-insensitive permeability of the inner membrane (open pores) of liver mitochondria as well as their monocarboxylic analogs.     

Mechanism of inhibition of beta-hexosaminidase B from human liver by short- and medium-chain monocarboxylic acids

Short- and medium-chain monocarboxylic acids showed an inhibitory effect on enzymatic activity of beta-hexosaminidase B (Hex B) when 4-methylumbelliferyl-2-acetamido-2-deoxyglucopyranoside (MU-GlcNAc) was used as substrate: 1. Two groups were distinguished according to the chain length of the monocarboxylic acids: the first was only constituted by acetic acid (C2) whereas the second group exhibited a broader chain length specificity for medium-chain monocarboxylic acids (between C6 and C9). 2. Both groups were reversible competitive inhibitors (Km = 0.52 +/- 0.15 mM; KiC2 = 21.4 +/- 3.0 mM; KiC7 = 3.4 +/- 0.5 mM). Competition experiments between C2 and C7 (as representent of medium-chain monocarboxylic acids group) demonstrated that these inhibitors were bound to different subsites. 3. Competition experiments between C2 and 2-acetamido-2-deoxy-D-galactonolactone (GalNAcLone) (a competitive inhibitor of lysosomal hexosaminidases) demonstrated that these two inhibitors were mutually exclusive, i.e. they were probably bound at the same subsite. This feature and the structural analogy of C2 with the acetyl residue of GalNAcLone (and of the saccharidic part of the substrate) suggested that C2 bound to the substrate site where the N-acetyl residue of the beta-N-acetyl hexosaminide was positioned. 4. The inhibitory effect of medium-chain monocarboxylic acids (C6 and C9) was dependent on their physical state. Below the critical micellar concentration (CMC), detected by a dye spectral shift method, no significant inhibition was detected, but as extensively reported using C7, an obvious inhibitory effect occurred at concentrations higher than CMC.(ABSTRACT TRUNCATED AT 250 WORDS)     

Transport of acetic acid in Zygosaccharomyces bailii: effects of ethanol and their implications on the resistance of the yeast to acidic environments.

Cells of Zygosaccharomyces bailii ISA 1307 grown in a medium with acetic acid, ethanol, or glycerol as the sole carbon and energy source transported acetic acid by a saturable transport system. This system accepted propionic and formic acids but not lactic, sorbic, and benzoic acids. When the carbon source was glucose or fructose, the cells displayed activity of a mediated transport system specific for acetic acid, apparently not being able to recognize other monocarboxylic acids. In both types of cells, ethanol inhibited the transport of labelled acetic acid. The inhibition was noncompetitive, and the dependence of the maximum transport rate on the ethanol concentration was found to be exponential. These results reinforced the belief that, under the referenced growth conditions, the acid entered the cells mainly through a transporter protein. The simple diffusion of the undissociated acid appeared to contribute, with a relatively low weight, to the overall acid uptake. It was concluded that in Z. bailii, ethanol plays a protective role against the possible negative effects of acetic acid by inhibiting its transport and accumulation. Thus, the intracellular concentration of the acid could be maintained at levels lower than those expected if the acid entered the cells only by simple diffusion.     

Lipid components of the seagrasses Posidonia australis and Heterozostera tasmanica as indicators of carbon source

The component hydrocarbons, sterols, alcohols, monocarboxylic, α, ω-dicarboxylic and ω-hydroxy acids of the seagrasses Posidonia australis and Heterozostera tasmanica and a sample of P. australis detritus are reported. The fresh leaves of P. australis and P. australis detritus are characterized by a distinctive distribution of solvent-extractable long-chain monocarboxylic, α, ω-dicarboxylic and ω-hydroxy acids. This distinctive pattern should enable these lipid components along with other distinctive components to be used as chemical markers of the seagrass P. australis. H. tasmanica is characterized by (1) higher relative concentrations of 16:2ω6 and 16:3ω3 than P. australis, (2) the absence of the distinctive distribution pattern of long-chain monocarboxylic and ω-hydroxy acids observed for P. australis, (3) the absence of α, ω-diacids and (4) a lower absolute concentration of ω-hydroxy acids than P. australis.     

Influence des lecithines dicarboxyliques sur les liposomes

We have investigated the effect of dicarboxylic lecithins on liposomes prepared from egg-lecithins and their incorporation into liposome preparation.In the presence of phospholipids, liposome cell permeability to glucose is five times higher than in controls.The incorporation of dicarboxylic lecithins into liposomes at a molar concentration of 50 molecules per cent reduces by 60% the amount of glucose trapped inside the liposomes.Measurements of ovelecithin-glutaryl-lecithin liposomes stability show that stable structures are formed at a molar concentration of 70 molecules per cent in the presence of cholesterol. On the contrary, the liposomes prepared in the absence of cholesterol are not stable at a molar concentration of 50 molecules per cent.Measurements of sedimentation of mixed dispersions of monocarboxylic lecithins and glutaryl-lecithins show that glutaryl-lecithins must be in the form of micelles to solubilize the phosphatidyl cholines.     

Two mire species respond differently to enhanced ultraviolet-B radiation: effects on biomass allocation and root exudation

Increased ultraviolet-B (UV-B) radiation arising from stratospheric ozone depletion may influence soil microbial communities via effects on plant carbon allocation and root exudation. Eriophorum angustifolium and Narthecium ossifragum plants, grown in peatland mesocosms consisting of Sphagnum peat, peat pore water and natural microbial communities, were exposed outdoors to enhanced UV-B radiation simulating 15% ozone depletion in southern Scandinavia for 8 wk. Enhanced UV-B increased rhizome biomass and tended to decrease the biomass of the largest root fraction of N. ossifragum and furthermore decreased dissolved organic carbon (DOC) and monocarboxylic acid concentration, which serves as an estimate of net root exudation, in the pore water of the N. ossifragum mesocosms. Monocarboxylic acid concentration was negatively related to the total carbon concentration of N. ossifragum leaves, which was increased by enhanced UV-B. By contrast, enhanced UV-B tended to increase monocarboxylic acid concentration in the rhizosphere of E. angustifolium and its root : shoot ratio. Microbial biomass carbon was increased by enhanced UV-B in the surface water of the E. angustifolium mesocosms. Increased UV-B radiation appears to alter below-ground biomass of the mire plants in species-specific patterns, which in turn leads to a change in the net efflux of root exudates.     

The utilization of short-chain monocarboxylic acids as carbon sources for the production of gamma-linolenic acid by Mucor strains in fed-batch culture

The Fischer-Tropsch reaction water, which contains C₂ to C₅ monocarboxylic acids, generated as a co-product of the Sasol industrial oil-from-coal process, constitutes a potential cheap carbon substrate for the production of gamma-linolenic acid (GLA) by selced Mucor species. Three strains of Mucor were each grown in an air-lift reactor operated in a fed-batch, pH-stat mode under N-limitation with a mixture of C₂ to C₅ monocarboxylic acids as both pH titrant and carbon source. The production of GLA from this substrate was evaluated. Growth typically resulted in the rapid assimilation of acetic, n-butyric and n-valeric acids. Although propionic, iso-butyric and iso-valeric acids were assimilated to varying degrees, these acids accumulated in the culture. Mucor circinelloides CBS 203.28 gave the best results in that it assimilated 36% to 100% of each acid, had a biomass yield coefficient of 0.3 (calculated on acids utilized), and contained 28% crude oil, 84% of which comprised neutral lipids with a GLA content of 14.4%, giving 33 mg GLA/g biomass. GLA accumulation coincided with a decrease in the stearic-acid content of the neutral-lipid fraction. The results were comparable with previous results obtained with acetic acid and glucose as sole carbon sources, demonstrating the feasibility of producing GLA from the above mixture of organic acids.     

Inhibitory effects of anions and active site amino acid sequence of chicken liver L-2-hydroxyacid oxidase A, a member of the FMN-dependent α-hydroxyacid oxidizing enzyme family

Monocarboxylic acids with aliphatic chains were found to be mixed inhibitors of chicken liver L-2-hydroxyacid oxidase A when L-2-hydroxy-4-methylthiobutanoic acid was used as the substrate. The finding that the binding affinity of the enzyme for monocarboxylic acids was directly proportional to the number of carbon atoms in the chain strongly suggests that in addition to the electrostatic interaction due to the carboxyl moiety, hydrophobic forces may also be involved in the binding affinity of monocarboxylic acids to the enzyme's active site. Oxalate, a dicarboxylic acid, also resulted in a mixed-type inhibition of chicken liver L-2-hydroxyacid oxidase A, and, surprisingly, its binding affinity to the enzyme was found to be quite high as compared with monocarboxylic acids. This is probably due to the fact that the two carboxyl groups of oxalate give rise to electrostatic interactions with the positively charged side chains of two adjacent residues in the polypeptide chain. The inhibitory effects of other dicarboxylic acids was found to decrease as the number of carbon atoms in the chain increased. Oxamate was found however to be a novel type of potent inhibitor of the enzyme. All in all, these kinetic studies and the amino acid sequence determination in the active site region after limited proteolysis of the polypeptide chain definitely establish that chicken liver NADH/FMN containing L-2-hydroxyacid oxidase A is a member of the FMN-dependent α-hydroxyacid oxidizing enzyme family.     

Irreversible inhibition of hexosaminidase C by medium-chain monocarboxylic acids and Triton X-100

The neutral beta-N-acetylhexosaminidase (hexosaminidase C) from human brain was partially purified (separated from lysosomal beta-N-acetylhexosaminidases by chromatography on a Con A-Sepharose column). Hexosaminidase C was inhibited by medium-chain fatty acids (monocarboxylic acids with chain-length between C6 and C9), whereas shorter-chain monocarboxylic acids showed no inhibitory effect. Studies on the inhibition mechanism showed an irreversible and pH-dependent inhibition which progresses with time and which is not reversed by the removal of fatty acids (by Bio-Beads SM-2). Similar inhibitory effects were also obtained using Triton X-100 (but not with homologous alkylamines). These results suggest that the hexosaminidase C inactivation is related to the hydrophobic properties of the inhibitor which acts as a denaturing agent mainly at acidic pH. The possibility has been discussed that this inactivation effect of monocarboxylic acid on hexosaminidase C could constitute a molecular model of the toxicity of medium-chain-length fatty acids.     

Partial molar volumes of some alpha-amino acids in aqueous sodium acetate solutions at 308.15 K

The apparent molar volumes V(2,phi) have been determined for glycine, DL-alpha-alanine, DL-alpha-amino-n-butyric acid, DL-valine and DL-leucine in aqueous solutions of 0.5, 1.0, 1.5 and 2.0 mol kg(-1) sodium acetate by density measurements at 308.15 K. These data have been used to derive the infinite dilution apparent molar volumes V(0)(2,phi) for the amino acids in aqueous sodium acetate solutions and the standard volumes of transfer, Delta(t)V(0), of the amino acids from water to aqueous sodium acetate solutions. It has been observed that both V(0)(2,phi) and Delta(t)V(0) vary linearly with increasing number of carbon atoms in the alkyl chain of the amino acids. These linear correlations have been utilized to estimate the contributions of the charged end groups (NH(3)(+), COO(-)), CH(2) group and other alkyl chains of the amino acids to V(0)(2,phi) and Delta(t)V(0). The results show that V(0)(2,phi) values for (NH(3)(+), COO(-)) groups increase with sodium acetate concentration, and those for CH(2) are almost constant over the studied sodium acetate concentration range. The transfer volume increases and the hydration number of the amino acids decreases with increasing electrolyte concentrations. These facts indicate that strong interactions occur between the ions of sodium acetate and the charged centers of the amino acids. The volumetric interaction parameters of the amino acids with sodium acetate were calculated in water. The pair interaction parameters are found to be positive and decreased with increasing alkyl chain length of the amino acids, suggesting that sodium acetate has a stronger dehydration effect on amino acids which have longer hydrophobic alkyl chains. These phenomena are discussed by means of the co-sphere overlap model.     

Dicarboxylic acids with limited numbers of hydrocarbons stabilize cell membrane and increase osmotic resistance in rat erythrocytes

We examined the effect of dicarboxylic acids having 0 to 6 hydrocarbons and their corresponding monocarboxylic or tricarboxylic acids in changing the osmotic fragility (OF) in rat red blood cells (RBCs). Malonic, succinic, glutaric and adipic acids, which are dicarboxylic acids with 1, 2, 3 and 4 straight hydrocarbons located between two carboxylic groups, decreased the OF in a concentration-dependent manner. Other long-chain dicarboxylic acids did not change the OF in rat RBCs. The benzoic acid derivatives, isophthalic and terephthalic acids, but not phthalic acid, decreased the OF in a concentration-dependent manner. Benzene-1,2,3-tricarboxylic acid, but not benzene-1,3,5-tricarboxylic acid, also decreased the OF in rat RBCs. On the other hand, monocarboxylic acids possessing 2 to 7 straight hydrocarbons and benzoic acid increased the OF in rat RBCs. In short-chain dicarboxylic acids, a limited number of hydrocarbons between the two carboxylic groups are thought to form a V- or U-shaped structure and interact with phospholipids in the RBC membrane. In benzene dicarboxylic and tricarboxylic acids, a part of benzene nucleus between the two carboxylic groups is thought to enter the plasma membrane and act on acyl-chain in phospholipids in the RBC membrane. For dicarboxylic and tricarboxylic acids, limited numbers of hydrocarbons in molecules are speculated to enter the RBC membrane with the hydrophilic carboxylic groups remaining outside, stabilizing the structure of the cell membrane and resulting in an increase in osmotic resistance in rat RBCs.     

镉在有机酸存在时对红壤中微生物生物量的影响

在预培红壤中加入定量的有机酸和不同浓度的Cd ,2 5℃培养 14d ,测定土壤微生物生物量C(Cmic)、N(Nmic) ,结果表明 ,存在有机酸时 ,土壤中Cmic和Nmic随着Cd浓度的增加而降低 ;Cmic/Nmic随着Cd浓度的增加而升高 .施加低分子量有机酸的土壤中 ,Cd浓度高于 2 5mg·kg-1土时 ,土壤中Cmic和Nmic均比未加有机酸时低 ,说明此时低分子量有机酸助长Cd的毒性 ,而Cd浓度低于 2 5mg·kg-1土时 ,土壤中Cmic和Nmic均比未加有机酸时高 ,说明此时低分子量有机酸可降低部分Cd的毒性 ;施加胡敏酸的土壤中 ,土壤中Cmic和Nmic均比未加有机酸时高 ,说明胡敏酸降低Cd的毒性并提供N源 .不含Cd时 ,加入有机酸导致土壤中Cmic和Nmic增加 ,其中胡敏酸最明显     

L-lactate transport in Ehrlich ascites-tumour cells.

Ehrlich ascites-tumour cells were investigated with regard to their stability to transport L-lactate by measuring either the distribution of [14C]lactate or concomitant H+ ion movements. The movement of lactate was dependent on the pH difference across the cell membrane and was electroneutral, as evidenced by an observed 1:1 antiport for OH- ions or 1:1 symport with H+ ions. 2. Kinetic experiments showed that lactate transport was saturable, with an apparent Km of approx. 4.68 mM and a Vmax. as high as 680 nmol/min per mg of protein at pH 6.2 and 37 degrees C. 3. Lactate transport exhibited a high temperature dependence (activation energy = 139 kJ/mol). 4. Lactate transport was inhibited competitively by (a) a variety of other substituted monocarboxylic acids (e.g. pyruvate, Ki = 6.3 mM), which were themselves transported, (b) the non-transportable analogues alpha-cyano-4-hydroxycinnamate (Ki = 0.5 mM), alpha-cyano-3-hydroxycinnamate (Ki = 2mM) and DL-p-hydroxyphenyl-lactate (Ki = 3.6 mM) and (c) the thiol-group reagent mersalyl (Ki = 125 muM). 5. Transport of simple monocarboxylic acids, including acetate and propionate, was insensitive to these inhibitors; they presumably cross the membrane by means of a different mechanism. 6. Experiments using saturating amounts of mersalyl as an "inhibitor stop" allowed measurements of the initial rates of net influx and of net efflux of [14C]lactate. Influx and efflux of lactate were judged to be symmetrical reactions in that they exhibited similar concentration dependence. 7. It is concluded that lactate transport in Ehrlich ascites-tumour cells is mediated by a carrier capable of transporting a number of other substituted monocarboxylic acids, but not unsubstituted short-chain aliphatic acids.     

Stimulation of pollen tube growthin vitro by dicarboxylic acids

Summary Effect of eight dicarboxylic acids and three monocarboxylic acids on pollen growth ofCamellia japonica was tested. While monocarboxylic acids inhibited pollen germination and pollen tube elongation, dicarboxylic acids, namely oxalic, succinic, suberic, adipic, sebacic, traumatic cis-1,2-cyclohexane dicarboxylic, and 3,3-diethyl glutaric acids stimulated pollen tube elongation stronger than indoleacetic acid.     

Specificity of the Na+-dependent monocarboxylic acid transport pathway in rabbit renal brush border membranes

The substrate specificity of a Na+-dependent transport pathway for L-lactate was studied in rabbit renal brush border membrane vesicles. Jmax for L-lactate transport was unaffected by the presence of a fixed concentration of two different short-chain monocarboxylic acids, while the apparent Kt(Ka) for L-lactate increased, and this is compatible with competitive inhibition. The inhibitor constants ("Ki"'s) for the transport pathway for the two solutes examined closely corresponded to the respective "Ki"'s derived from a Dixon plot. A broad range of compounds were then tested as potential inhibitors of L-lactate transport, and the "Ki"'s thereby derived yielded specific information regarding optimal substrate recognition by the carrier. A single carboxyl group is an absolute requirement for recognition, and preference is given to 3 to 6 C chain molecules. Addition of ketone, hydroxyl and, particularly, amine groups at any carbon position, diminishes substrate-carrier interaction. Intramolecular forces, notably the inductive effects of halogens, may play a role in enhancing substrate-carrier interaction; however, no correlation was found between pKa and "Ki" for the substrates examined. We conclude that a separate monocarboxylic acid transport pathway, discrete from either the D-glucose, alpha or beta neutral amino-acid, or dicarboxylic acid carriers, exists in the renal brush border, and this handles a broad range of monocarboxylates.     

Functional analysis of a mammalian odorant receptor subfamily

Phylogenetic analysis groups mammalian odorant receptors into two broad classes and numerous subfamilies. These subfamilies are proposed to reflect functional organization. Testing this idea requires an assay allowing detailed functional characterization of odorant receptors. Here we show that a variety of Class I and Class II mouse odorant receptors can be functionally expressed in Xenopus laevis oocytes. Receptor constructs included the N-terminal 20 residues of human rhodopsin and were co-expressed with Galphaolf and the cystic fibrosis transmembrane regulator to allow electrophysiological measurement of receptor responses. For most mouse odorant receptors tested, these conditions were sufficient for functional expression. Co-expression of accessory proteins was required to allow functional surface expression of some mouse odorant receptors. We used this assay to examine the receptive ranges of all members of the mouse odorant receptor 42 (MOR42) subfamily. MOR42-1 responded to dicarboxylic acids, preferring a 10-12 carbon chain length. MOR42-2 responded to monocarboxylic acids (7-10 carbons). MOR42-3 responded to dicarboxylic acids (8-10 carbons) and monocarboxylic acids (10-12 carbons). Thus, the receptive range of each receptor was unique. However, overlap between the individual receptive ranges suggests that the members of this subfamily form one contiguous subfamily receptive range, suggesting that odorant receptor subfamilies do constitute functional units.