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PubMed2002

Characteristics of L-lactic acid transport in basal membrane vesicles of human placental syncytiotrophoblast.

The characteristics of L-lactic acid transport across the trophoblast basal membrane were investigated and compared with those across the brush-border membrane by using membrane vesicles isolated from human placenta. The uptake of L-[(14)C]lactic acid into basal membrane vesicles was Na(+) independent, and an uphill transport was observed in the presence of a pH gradient ([H(+)](out) > [H(+)](in)). L-[(14)C]lactic acid uptake exhibited saturation kinetics with a K(m) value of 5.89 +/- 0.68 mM in the presence of a pH gradient. p-Chloromercuribenzenesulfonate and alpha-cyano-4-hydroxycinnamate inhibited the initial uptake, whereas phloretin or 4,4'-diisothiocyanostilbene-2,2'-disulfonate did not. Mono- and dicarboxylic acids suppressed the initial uptake. In conclusion, L-lactic acid transport in the basal membrane is H(+) dependent and Na(+) independent, as is also the case for the brush-border membrane transport, and its characteristics resemble those of monocarboxylic acid transporters. However, there were several differences in the effects of inhibitors between basal and brush-border membrane vesicles, suggesting that the transporter(s) involved in L-lactic acid transport in the basal membrane of placental trophoblast may differ from those in the brush-border membrane.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2006

Production of lactic acid with loofa sponge immobilized Rhizopus oryzae RBU2-10.

Lactic acid production by Rhizopus oryzae RBU2-10 immobilized in loofa sponge was evaluated. Shape and texture of loofa sponge, which was obtained from the mature dried fruit of Luffa cylindrica, remained intact after its treatment with buffers of varying pH and following its repeated autoclaving for up to four cycles (121 degrees C, 20 min per cycle). The medium having four pieces of loofa sponge (1.008 cm(3)) per 100 ml medium and inoculated with 3 x1 0(6) spores ml(-1) resulted maximum production (80.75 g l(-1)) of lactic acid in 48 h of fermentation. Repeated batch fermentation for lactic acid production could be carried out for 10 cycles. Remarkably higher levels of productivity (1.66-1.84 g l(-1)h(-1)) was obtained during first five cycles of fermentation with a maximum productivity (1.84 g l(-1)h(-1)) obtained during third cycle of fermentation.

Animals↗
PubMed1996

The proteolytic systems of lactic acid bacteria.

Proteolysis in dairy lactic acid bacteria has been studied in great detail by genetic, biochemical and ultrastructural methods. From these studies the picture emerges that the proteolytic systems of lactococci and lactobacilli are remarkably similar in their components and mode of action. The proteolytic system consists of an extracellularly located serine-proteinase, transport systems specific for di-tripeptides and oligopeptides (> 3 residues), and a multitude of intracellular peptidases. This review describes the properties and regulation of individual components as well as studies that have led to identification of their cellular localization. Targeted mutational techniques developed in recent years have made it possible to investigate the role of individual and combinations of enzymes in vivo. Based on these results as well as in vitro studies of the enzymes and transporters, a model for the proteolytic pathway is proposed. The main features are: (i) proteinases have a broad specificity and are capable of releasing a large number of different oligopeptides, of which a large fraction falls in the range of 4 to 8 amino acid residues; (ii) oligopeptide transport is the main route for nitrogen entry into the cell; (iii) all peptidases are located intracellularly and concerted action of peptidases is required for complete degradation of accumulated peptides.

Amino Acid Sequence↗
PubMed2002

Combined effects of lactic acid and nisin solution in reducing levels of microbiological contamination in red meat carcasses.

Changes in bacterial counts on beef carcasses at specific points during slaughter and fabrication were determined, and the effectiveness of nisin, lactic acid, and a combination of the lactic acid and nisin in reducing levels of microbiological contamination was assessed. Swab samples were obtained from the surfaces of randomly selected beef carcasses. Carcasses were swabbed from the neck, brisket, and renal site after skinning, splitting, and washing. Treatments involving lactic acid (1.5%), nisin (500 IU/ml), or a mixture of nisin and lactic acid were applied after the neck area was washed. A control group was not sprayed. Results indicated that the highest prevalence of aerobic plate counts (APCs), total coliforms, and Escherichia coli was found in the neck site after splitting, and the lowest level of microbial contamination was found after skinning. Washing with water did not significantly reduce the bacterial load. The largest reduction in APCs, total coliforms, and E. coli occurred on carcasses treated with a mixture of nisin and lactic acid. A mixture of nisin and lactic acid can be applied to beef carcasses through spray washing and can reduce bacterial populations by 2 log units.

Animals↗
PubMed2004

Plasma D-lactic acid level: a useful marker to distinguish perforated from acute simple appendicitis.

Early diagnosis of perforated appendicitis is important for reducing morbidity rates. The aim of this study was to determine the value and utility of plasma D-lactic acid levels in identifying the type of appendicitis. In this clinical study, plasma D-lactic acid levels were assessed in 44 consecutive paediatric patients (23 with acute appendicitis, 21 with perforated appendicitis) before laparotomy. D-lactic acid levels were determined by an enzymatic spectrophotometric technique using a D-lactic acid dehydrogenase kit. Patients with perforated appendicitis had higher D-lactic acid levels (3.970 +/- 0.687 mg/dL) than patients in the control group (0.478 +/- 0.149 mg/dL) and patients with acute appendicitis (1.409 +/- 0.324 mg/dL; p < 0.05). For a plasma D-lactic acid level greater than 2.5 mg/dL, the sensitivity and specificity of the D-lactic acid assay were 96% and 87%, respectively. The positive predictive value was 87%, the negative predictive value was 96%, and the diagnostic value was 91%. These results suggest that the measurement of plasma D-lactic acid levels may be a useful adjunct to clinical and radiological findings in distinguishing perforated from acute non-perforated appendicitis in children.

Acute Disease↗
PubMed2004

Lactic acid root canal irrigation for dowel and core treatment: a pilot study.

STATEMENT OF PROBLEM: Various solutions used to irrigate root canals and remove the smear layer prior to obturation of a root canal affect surface topography and may influence the sealing ability of endodontic therapy, retention of endodontic dowels, and restoration survival. PURPOSE: This study microscopically evaluated the effects of varying dilutions of lactic acid on the removal of the smear layer created by hand instrumentation in the coronal, middle, and apical thirds of root canals. Lactic acid solution was compared with other solutions deemed to be suitable root canal irrigants. MATERIAL AND METHODS: Thirty-five extracted human single-rooted teeth were manually shaped with files (K-flex) using the step-back technique. ISO size 50 files were used as master apical files. The teeth were equally divided into 7 test groups according to the canal irrigant used. While the teeth were cleaned and shaped, the root canals were irrigated with 3 mL of 1 of the following irrigants between each file size and as a final flush: no irrigant (control), 5% hydrogen peroxide, 5% sodium hypochlorite, a combination of 5% hydrogen peroxide and 5% sodium hypochlorite, 15% ethylenediaminotetraacetic acid (EDTA), 10% lactic acid, or 20% lactic acid. Specimens were critical-point dried and freeze-fractured for scanning electron microscope analysis at 3 sites (cervical, middle, and apical). RESULTS: Scanning electron micrographs showed no differences between the cervical, middle, and apical levels of the specimens tested with any of the irrigants used. Also, no difference was found within individual groups. The surface of the control specimens had a smear layer that contained smear plug material. Hydrogen peroxide and sodium hypochlorite used separately and in combination consistently produced surfaces with a smear layer. Lactic acid solutions and EDTA, however, removed the dentinal smear layer but left remnants of smear plugs in the dentinal tubules. CONCLUSIONS: Within the limitations of this study, differences related to the type of irrigant used were found in the characteristics of the pulpal dentin surface. Although lactic acid effectively cleaned and smoothed the dentin walls of root canals of endodontically prepared teeth, further study is needed before this procedure can be recommended for routine clinical use.

Chelating Agents↗
PubMed1989

In vitro effects of gossypol and lactic acid on rat uterus and ovary during implantation and antiimplantation.

ATPase activity of uterus and ovary was markedly elevated in presence of gossypol and decreased in presence of lactic acid indicating activation and inhibition of energy metabolism by gossypol and lactic acid respectively. The elevated levels of glycogen in uterus indicate inhibition of glycogenolysis as supported by phosphorylase activity. Whereas in ovary the glycogen depletion indicates activation of glycogenolysis supported by phosphorylase activity. The activity levels of aldolase and G-6-PDH decreased in the uterus in presence of gossypol and increased in presence of lactic acid. The same were elevated in ovary indicating the activation of hexose mono and diphosphate pathways. Lactic acid accumulated in presence of both gossypol and lactic acid with a depletion in level of pyruvic acid in both the tissues. This situation in the uterus indicates the condition of anti-implantation in presence of both gossypol and lactic acid. The NAD-LDH activity was inhibited in presence of gossypol and activated in presence of lactic acid in both tissues.

Animals↗
PubMed1996

Combined effects of packaging atmosphere and lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat 4 degrees C.

The effect of lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat stored under refrigeration and various gas environments was investigated. Frozen crayfish tail meat was thawed overnight, autoclaved, cooled, and inoculated with approximately 4 log colony-forming units (CFU) of a mixed-strain (Scott A and F5027) L. monocytogenes culture per gram of meat. Inoculated samples were blended with 0, 0.5, 1.0, 1.5, or 2.0% lactic acid and packaged under air, vacuum, or modified atmosphere (74.8% CO2, 10.4% O2, and 14.8% N2) and stored at 4 degrees C for 20 days. Results demonstrated that modified atmosphere packaging inhibited the growth of L. monocytogenes more than air and vacuum packaging at 0 and 1% lactic acid. Microbial counts declined steadily in crayfish tail meat treated with 2% lactic acid, with no differences among the packaging atmospheres. The lag phase was extended by 8 days in samples treated with 1% lactic acid and modified atmosphere compared to that in air or vacuum packaging. Overall, the combination of lactic acid and modified atmosphere had the greatest potential to prevent growth of L. monocytogeines.

Animals↗
PubMed1974

Lactic acid translocation: terminal step in glycolysis by Streptococcus faecalis.

Streptococcus faecalis obtains metabolic energy chiefly from the conversion of glucose to lactic acid; the present experiments deal with the mechanism of lactic acid translocation across the cytoplasmic membrane. Efflux of [(14)C]lactate from preloaded cells was accelerated by raising the external pH, and also by the ionophores nigericin and valinomycin. These results suggest that lactate leaves the cell by an electroneutral process, presumably as lactic acid. Further evidence was obtained by studying the entry of [(14)C]lactate into nonmetabolizing cells. It appears that the membrane is essentially impermeable to the lactate anion, but allows passage of lactic acid. The most persuasive evidence is that, upon establishment of a pH gradient such that the cytoplasm was alkaline, l-[(14)C]lactate accumulated in the cells against the concentration gradient. Accumulation was transient, and dissipated in parallel with the collapse of the pH gradient. The concentration gradient attained at the peak was a function of the pH difference. Ionophores which are known to collapse a pH gradient, such as nigericin and valinomycin, abolished accumulation of l-lactate. We infer that lactic acid translocation, whether into the cells or outward, is an electroneutral process and for that reason the distribution of lactic acid across the membrane is a function of the pH of cytoplasm and medium. The specificity of translocation and its kinetic parameters suggest that it is mediated by a carrier of low specificity.

Carbon Radioisotopes↗
PubMed2005

Gene cloning and molecular characterization of an extracellular poly(L-lactic acid) depolymerase from Amycolatopsis sp. strain K104-1.

We have isolated a polylactide or poly(L-lactic acid) (PLA)-degrading bacterium, Amycolatopsis sp. strain K104-1, and purified PLA depolymerase (PLD) from the culture fluid of the bacterium. Here, we cloned and expressed the pld gene encoding PLD in Streptomyces lividans 1326 and characterized a recombinant PLD (rPLD) preparation. We also describe the processing mechanism from nascent PLD to mature PLD. The pld gene encodes PLD as a 24,225-Da polypeptide consisting of 238 amino acids. Biochemical and Western immunoblot analyses of PLD and its precursors revealed that PLD is synthesized as a precursor (prepro-type), requiring proteolytic cleavage of the N-terminal 35-amino-acid extension including the 26-amino-acid signal sequence and 9-residue prosequence to generate the mature enzyme of 20,904 Da. The cleavage of the prosequence was found to be autocatalytic. PLD showed about 45% similarity to many eukaryotic serine proteases. In addition, three amino acid residues, H57, D102, and S195 (chymotrypsin numbering), which are implicated in forming the catalytic triad necessary for cleavage of amide bond of substrates in eukaryotic serine proteases, were conserved in PLD as residues H74, D111, and S197. The G193 residue (chymotrypsin numbering), which is implicated in forming an oxyanion hole with residue S195 and forms an important hydrogen bond for interaction with the carbonyl group of the scissile peptide bond, was also conserved in PLD. The functional analysis of the PLD mutants H74A, D111A, and S197A revealed that residues H74, D111, and S197 are important for the depolymerase and caseinolytic activities of PLD and for cleavage of the prosequence from pro-type PLD to form the mature one. The PLD preparation had elastase activity which was not inhibited by 1 mM elastatinal, which is 10 times higher than needed for complete inhibition of porcine pancreatic elastase. The rPLD preparation degraded PLA with an average molecular mass of 220 kDa into lactic acid dimers through lactic acid oligomers and finally into lactic acid. The PLD preparation bound to high polymers of 3-hydoxybutyrate, epsilon-caprolacton, and butylene succinate as well as PLA, but it degraded only PLA.

3-Hydroxybutyric Acid↗
PubMedDate not supplied

[Adhesive properties of lactic acid bacteria and methods of their investigation].

Current status of the question concerning the study of lactic acid bacteria adhesion process is presented in the review. Lactic acid bacteria colonization capacity allows them to form monolayers, to join the mucosal microbiota, to become a component of ecological barrier, which protects an organism from penetration of pathogens. The available methods of investigation of adhesive properties such as microbiological, light and electron microscopy, biophysical, mathematical and others are presented in detail. The works concerning lactic acid bacteria adhesines nature and factors that influence the exhibition of adhesion have been analyzed. Studies concerning the mechanisms responsible for attachment of cells of lactic acid bacteria to the surface of different types of epithelium and substances that are products of endothelium secretion are analyzed. Questions on adhesion specificity and factors that inhibit its exhibition are considered. The chart of current concept of the adhesive process and factors that reciprocally affect the exhibition of lactic acid bacteria adhesion properties is developed and presented by the authors.

Adhesins, Bacterial↗
PubMed2002

Molecular typing techniques to characterize the development of a lactic acid bacteria community on vacuum-packaged beef.

The development of a community of lactic acid bacteria from vacuum-packaged beef was investigated during a 6-week storage trial at 2 degrees C. The lactic acid bacteria population was monitored by using molecular techniques to identify a random sample of isolates at biweekly intervals during the storage trial. The polymerase chain reaction and a randomly amplified polymorphic DNA technique were used to identify and distinguish populations of lactic acid bacteria that developed during the storage trial. At week 0, the population of lactic acid bacteria was 3.5 log cfu/120 cm2 and by week 6, the population reached a maximum of 7.6 log cfu/120 cm2. A sampling from the week 0 population indicated a mixed community of Lactobacillus curvatus, Lactobacillus sakei and Leuconostoc spp. However, the sampling from week 6 indicated the population composition had changed to one where a single Leuconostoc strain predominated. This strain demonstrated antagonism towards the growth of other lactic acid bacteria isolated during the study. Additionally, the strain inhibited the growth of foodborne pathogens Escherichia coli O157:H7 and Listeria monocytogenes. DNA sequence data from the 16S rRNA gene suggested that the isolate may be a Leuconostoc gelidum strain.

Animals↗
PubMed2003

Production of optically pure D-lactic acid in mineral salts medium by metabolically engineered Escherichia coli W3110.

The resistance of polylactide to biodegradation and the physical properties of this polymer can be controlled by adjusting the ratio of L-lactic acid to D-lactic acid. Although the largest demand is for the L enantiomer, substantial amounts of both enantiomers are required for bioplastics. We constructed derivatives of Escherichia coli W3110 (prototrophic) as new biocatalysts for the production of D-lactic acid. These strains (SZ40, SZ58, and SZ63) require only mineral salts as nutrients and lack all plasmids and antibiotic resistance genes used during construction. D-Lactic acid production by these new strains approached the theoretical maximum yield of two molecules per glucose molecule. The chemical purity of this D-lactic acid was approximately 98% with respect to soluble organic compounds. The optical purity exceeded 99%. Competing pathways were eliminated by chromosomal inactivation of genes encoding fumarate reductase (frdABCD), alcohol/aldehyde dehydrogenase (adhE), and pyruvate formate lyase (pflB). The cell yield and lactate productivity were increased by a further mutation in the acetate kinase gene (ackA). Similar improvements could be achieved by addition of 10 mM acetate or by an initial period of aeration. All three approaches reduced the time required to complete the fermentation of 5% glucose. The use of mineral salts medium, the lack of antibiotic resistance genes or plasmids, the high yield of D-lactate, and the high product purity should reduce costs associated with nutrients, purification, containment, biological oxygen demand, and waste treatment.

Acetates↗
PubMed2003

Metabolic engineering of Lactobacillus fermentum for production of mannitol and pure L-lactic acid or pyruvate.

For production of mannitol in combination with pure L-lactic acid or pyruvate, the D- and L-lactate dehydrogenase genes (ldhD and ldhL) of a mannitol-producing Lactobacillus fermentum strain were cloned and stepwise inactivated. For inactivation of both ldh genes by a gene replacement technique, deletion constructs removing a 0.4-kb fragment from the promoter and the 5' end region of the ldh genes were used. The first inactivation mutant, designated L. fermentum GRL1030, carried the deletion in ldhD (DeltaldhD). A double mutant, DeltaldhD-DeltaldhL, was constructed by the inactivation of the ldhL gene of strain GRL1030, resulting in strain L. fermentum GRL1032. The correctness of the both mutants was confirmed at the DNA level by polymerase chain reaction, as shown by the absence of ldh transcripts by northern blotting and as a lack of the corresponding enzyme activity. In bioreactor cultivations, the single mutant GRL1030 produced mannitol and L-lactic acid as expected. Mannitol and lactic acid yields and productivities were practically unaffected by deletion of the ldhD gene. The double mutant GRL1032 produced mannitol and pyruvate as expected. However, although the yield of mannitol from fructose remained high, its volumetric productivity was reduced. The double mutation negatively affected the glucose consumption rate, resulting in reduced cellular growth. In addition to pyruvate, the double mutant produced 2,3-butanediol. More surprisingly, some lactic acid was still produced.

Bioreactors↗
PubMed1994

Kinetics and Modeling of Lactic Acid Production by Lactobacillus plantarum.

An unstructured model was developed to describe bacterial growth, substrate utilization, and lactic acid production by Lactobacillus plantarum in cucumber juice. Significant lactic acid production occurred during growth, as well as stationary phases. The percentage of acid produced after growth ceased was a function of the medium composition. Up to 51% of the lactic acid was produced after growth ceased when NaCl was not present in the medium, whereas not more than 18% of the total lactic acid was produced after the growth ceased in presence of NaCl, probably because of an increase in the cell death rate. An equation relating the specific death rate and NaCl concentration was developed. With the kinetic model proposed by R. Luedeking and E. L. Piret (J. Biochem. Microbiol. Technol. Eng. 1:393-412, 1958) for lactic acid production rate, the growth-associated and non-growth-associated coefficients were determined as 51.9 (+/-4.2) mmol/g of cells and 7.2 (+/-0.9) mmol/g of cells h respectively. The model was demonstrated for batch growth of L. plantarum in cucumber juice. Mathematical simulations were used to predict the influence of variations in death rate, proton concentration when growth ceased, and buffer capacity of the juice on the overall fermentation process.

Journal Article↗
PubMed1998

Effect of L-lactic acid on calcium absorption in rats fed omeprazole.

We examined the effect of L-lactic acid on calcium absorption in male Wistar rats made achlorhydric by dietary omeprazole, a proton pump inhibitor. The dietary omeprazole intake (0.03 g/100 g of diet) increased the gastric pH and decreased the apparent calcium absorption ratio. Dietary famotidine (0.03 g/100 g of diet), an H2-receptor antagonist, and lower doses of omeprazole (0.005 or 0.01 g/100 g of diet) did not affect the gastric pH or the calcium absorption. In a second experiment, dietary lactic acid (0.5, 1.0, or 2.5 g/100 g of diet) increased the intestinal calcium absorption dose dependently in rats fed omeprazole (0.03 g/100 g of diet). The gastric pH was significantly decreased only in the rats fed higher doses of lactic acid (1.0, or 2.5 g/100 g of diet). In a third experiment, a dietary sour milk beverage containing lactic acid (0.5 g/100 g of diet) increased the intestinal calcium absorption, but did not affect the gastric pH in rats fed omeprazole (0.03 g/100 g of diet). Although the significance of gastric acid in terms of overall calcium absorption is not known, under the present experimental conditions, the inhibition of gastric acid secretion by dietary omeprazole decreased the apparent calcium absorption, and the dietary lactic acid prevented the calcium absorption in rats fed omeprazole.

Animals↗
PubMed1981

Effect of CO2 and lactic acid on intracellular pH of ascites tumor cells.

The effect of CO2 and of lactic acid (L.A.) on the extracellular (pHe) and intracellular pH (pHi) of ascites tumor cells (DS-carcinosarcoma) in rats was studied by in vitro equilibration of ascites with CO2 and alteration of lactic acid concentration. pHi was determined by the distribution of DMO. The effects of lactic acid and CO2 on pH were additive and could be expressed as pHe = 8.872 - 0.745 logPCO2 - 0.0355 (L.A.) (R = 0.867, n = 201) pHi = 8.218 - 0.436 logPCO2 - 0.0275 (L.A.) (R = 0.812, n = 143) delta pHi/delta pHe was dependent on the way pHe was changed: If the change in pHe was due to lactic acid, delta pHi/delta pHe was 0.91; if it was due to CO2 delta pHi/delta pHe was 0.625. pHi exceeded pHe if either PCO2 and/or the concentration of lactic acid was raised above a critical level. The results render it questionable to predict intracellular pH values within solid tumor from pH measurements within the extracellular fluid.

Animals↗
PubMed2006

Efficacy of lactic acid against Listeria monocytogenes attached to poultry skin during refrigerated storage.

AIMS: The aim of this study was to evaluate the effect of lactic acid washing on the growth of Listeria monocytogenes on poultry legs stored at 4 degrees C for 7 days. METHODS AND RESULTS: Fresh inoculated chicken legs were dipped into either a 0.11, 0.22 mol l(-1) or 0.55 mol l(-1) lactic acid solution for 5 min or distilled water (control). Surface pH values, sensorial characteristics and L. monocytogenes, mesophiles and pychrotrophs counts were evaluated after treatment (day 0) and after 1, 3, 5 and 7 days of storage at 4 degrees C. Legs washed with 0.55 mol l(-1) lactic acid for 5 min showed a significant (P < 0.05) inhibitory effect on L. monocytogenes compared with control legs, being about 1.74 log units lower in the first ones than in control legs after 7 days of storage. Sensory quality was not adversely affected by lactic acid, with the exception of colour. CONCLUSIONS: Treatments with 0.55 mol l(-1) lactic acid reduced bacterial growth and preserved reasonable sensorial quality after storage at 4 degrees C for 7 days. However, it was observed a reduction in the colour score within 1 day post-treatment with 0.55 mol l(-1) lactic. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that, while lactic acid did reduce populations of L. monocytogenes on poultry, it did not completely inactivate the pathogen. The application of lactic acid may be used as an additional hurdle contributing to extend the shelf-life of raw poultry.

Animals↗