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PubMed2005

Effect of expressing polyhydroxybutyrate synthesis genes (phbCAB) in Streptococcus zooepidemicus on production of lactic acid and hyaluronic acid.

Hyaluronic acid (HA) has been industrially produced using the gram-positive bacterium Streptococcus zooepidemicus. Large amount of lactic acid formation was one of the important factors that restricted cell growth and HA productivity and lowered the substrate to HA conversion efficiency in a fermentor. In this study, polyhydroxybutyrate (PHB) synthesis genes (phbCAB) of Ralstonia eutropha were cloned from the plasmid pBHR68 and were inserted into the plasmid pEU308, an expression vector for gram-positive bacteria. The plasmid was transformed into S. zooepidemicus by electroporation. beta-Ketothiolase (PhbA), acetoacetyl-CoA reductase (PhbB), and polyhydroxyalkanoate (PHA) synthase (PhbC) activity assays were carried out to demonstrate the expression of these genes. The PhbA and PhbB activities were 3.13 and 1.23 U mg(-1), respectively. No PhbC activities were detected. In shake flask studies, there was no obvious difference between the wild-type and recombinant S. zooepidemicus harboring phbCAB genes in terms of lactic acid and HA formation. However, in fermentor studies, the recombinant produced only 40 g L(-1) lactic acid and 7.5 g L(-1) HA, whereas the wild type produced 65 g L(-1) lactic acid and 5.5 g L(-1) HA. These results suggested that expression of phbCAB genes in S. zooepidemicus could help regulate HA production metabolism. Because the lactic acid formation in S. zooepidemicus was sensitive to cellular oxidation/reduction potential, it is proposed that the PHB synthesis pathway could act as a regulator to adjust the cellular oxidation/reduction potential. This is the first study demonstrating that PHA synthesis related to energy and carbon metabolism could be employed as a pathway to regulate other cellular metabolism and possibly to regulate the production of other metabolic products.

Acetyl-CoA C-Acyltransferase↗
PubMed2006

Interaction effects of lactic acid and acetic acid at different temperatures on ethanol production by Saccharomyces cerevisiae in corn mash.

The combined effects of lactic acid and acetic acid on ethanol production by S. cerevisiae in corn mash, as influenced by temperature, were examined. Duplicate full factorial experiments (three lactic acid concentrations x three acetic acid concentrations) were performed to evaluate the interaction between lactic and acetic acids on the ethanol production of yeast at each of the three temperatures, 30, 34, and 37 degrees C. Corn mash at 30% dry solids adjusted to pH 4 after lactic and acetic acid addition was used as the substrate. Ethanol production rates and final ethanol concentrations decreased (P<0.001) progressively as the concentration of combined lactic and acetic acids in the corn mash increased and the temperature was raised from 30 to 37 degrees C. At 30 degrees C, essentially no ethanol was produced after 96 h when 0.5% w/v acetic acid was present in the mash (with 0.5, 2, and 4% w/v lactic acid). At 34 and 37 degrees C, the final concentrations of ethanol produced by the yeast were noticeably reduced by the presence of 0.3% w/v acetic acid and >or=2% w/v lactic acid. It can be concluded that, as in previous studies with defined media, lactic acid and acetic acid act synergistically to reduce ethanol production by yeast in corn mash. In addition, the inhibitory effects of combined lactic and acetic acid in corn mash were more apparent at elevated temperatures.

Acetic Acid↗
PubMed1993

Energy transduction in lactic acid bacteria.

In the discovery of some general principles of energy transduction, lactic acid bacteria have played an important role. In this review, the energy transducing processes of lactic acid bacteria are discussed with the emphasis on the major developments of the past 5 years. This work not only includes the biochemistry of the enzymes and the bioenergetics of the processes, but also the genetics of the genes encoding the energy transducing proteins. The progress in the area of carbohydrate transport and metabolism is presented first. Sugar translocation involving ATP-driven transport, ion-linked cotransport, heterologous exchange and group translocation are discussed. The coupling of precursor uptake to product product excretion and the linkage of antiport mechanisms to the deiminase pathways of lactic acid bacteria is dealt with in the second section. The third topic relates to metabolic energy conservation by chemiosmotic processes. There is increasing evidence that precursor/product exchange in combination with precursor decarboxylation allows bacteria to generate additional metabolic energy. In the final section transport of nutrients and ions as well as mechanisms to excrete undesirable (toxic) compounds from the cells are discussed.

Adenosine Triphosphate↗
PubMed1991

Effect of lactic acid producing bacteria on the human intestinal microflora during ampicillin treatment.

20 healthy volunteers participated in a double blind study concerning the effect of lactic acid producing bacteria on the intestinal microflora during ampicillin treatment. 10 volunteers received 500 mg ampicillin tablets t.i.d. together with capsules containing lactic acid producing bacteria (Lactobacillus acidophilus and Bifidobacterium bifidum) for 7 days, and the other 10 volunteers were given 500 mg ampicillin tablets together with placebo capsules t.i.d. for 7 days. Both groups of volunteers continued the intake of the capsules t.i.d. for another 14 days after the ampicillin administration had been completed. The number of enterococci, streptococci and corynebacteria decreased during ampicillin administration but returned to normal levels after 14 days. Yeasts increased during the antibiotic treatment but returned to the same levels as before treatment within 14 days. Escherichia coli strains were suppressed in most volunteers during ampicillin administration. The numbers of anaerobic gram-positive cocci and rods decreased in most subjects during ampicillin treatment but were normalized within 2 weeks. Bacteroides strains were recovered in higher numbers in the lactic acid producing bacteria group compared to the placebo group. The volunteers receiving lactic acid producing bacteria were recolonized slightly faster than those having placebo. There were adverse effects observed in 3 subjects receiving ampicillin plus placebo. In the lactic acid producing bacteria group, one subject had diarrhoea on day 3 to on day 3 to day 7.

Adult↗
PubMed1994

Aqueous humour lactic acid and proteins in eyes with iris neovascularization.

The content of lactic acid and proteins in the aqueous humour of eyes with neovascularization of the iris (NVI) of various etiology was determined. The results were analysed for any possible correlation with the angiogenesis observed in such eyes. Aqueous humour from patients with immature senile cataracts served as the control. The total protein concentration in cases of NVI (mean +/- SD, 345.7 +/- 161.0 mg/dl; n = 15) was significantly raised (P < 0.001) as compared with the control values (mean +/- SD, 30.91 +/- 14.95 mg/dl; n = 12). On the other hand, the concentration of lactic acid in cases of NVI (mean +/- SD, 45.97 +/- 15.5 mg/dl; n = 13) was significantly lower (P < 0.01) as compared with the controls (mean +/- SD, 65.86 +/- 10.9 mg/dl; n = 8). These findings show that angiogenesis is not associated with a raised level of lactic acid in the aqueous humour, whereas the elevated protein values are suggestive of a breakdown of the blood-aqueous barrier.

Adult↗
PubMed2000

Sustained drug release characteristics of biodegradable composite poly(d,l)lactic acid-poly(l)lactic acid microcapsules containing ciprofloxacin.

Ciprofloxacin polylactic microcapsules were prepared by the phase separation process. Two types of polylactic acid, poly(d,l)lactic acid and poly(l)lactic acid were combined as membrane materials to prevent the aggregation which happened frequently in the phase separation process. The polymer compositions of the microcapsules can influence the release rate of Ciprofloxacin. The optimal release rate of the drug can be obtained by modifying microcapsule compositions. Poly(d,l)lactic acid is superior in slowing the rate of drug release than poly(l)lactic acid. However, poly(l)lactic acid is necessary in the preparation of the microcapsules to prevent aggregation.

Anti-Infective Agents↗
PubMed1980

Myocardial lactic acid balance after left ventriculography.

In order to study the effects of contrast media on myocardial metabolism the percentage of lactic acid extraction during left ventricular angiography has been determined in 16 normal patients (Group I), and in 29 patients with coronary artery disease (Group II). In addition, the lactic acid uptake was evaluated after atrial pacing (in 10 patients of Group I and in 13 patients of Group II) and after aortography (in 6 patients of Group I and in 9 patients of Group II). Normal patients usually did not show a myocardial lactic acid production after left ventriculography while the patients with coronary artery disease showed a net lactic acid production in the coronary sinus, which was similar to that obtained after atrial pacing stimulation and after aortography. These results indicate that ventriculography can be a useful technique to assess the kinetics of lactic acid uptake or release into the coronary sinus in patients with coronary artery disease and, in this respect could be useful for studying myocardial metabolism in aortic valve disease and to assess the toxicity of different contrast media.

Adult↗
PubMed1994

Isolation from food sources, of lactic acid bacteria that produced antimicrobials.

The potential of lactic acid bacteria, isolated from a variety of foods, to inhibit indicators representative of spoilage and pathogenic bacteria associated with food products was examined. Fruit and vegetables were a poor source of lactic acid bacteria but large numbers were readily isolated on MRS agar from cheese, milk and meat samples. Approximately 1000 isolates from each of the food samples were examined by the deferred antagonism procedure to determine their ability to inhibit Staphylococcus aureus, Listeria innocua and Pseudomonas fragi. Listeria innocua was the bacterium predominantly inhibited by isolates from the cheese, milk and meats, but antagonism was also observed to a lesser extent against the other indicators. The only inhibition observed for isolates from vegetable material was directed against Staph. aureus. The majority of inhibitor producers were effective against only one of the indicators but a small number were isolated which inhibited two or three.

Animals↗
PubMed2005

Optimization of lactic acid production in SSF by Lactobacillus amylovorus NRRL B-4542 using Taguchi methodology.

Lactic acid production parameter optimization using Lactobacillus amylovorus NRRL B-4542 was performed using the design of experiments (DOE) available in the form of an orthogonal array and a software for automatic design and analysis of the experiments, both based on Taguchi protocol. Optimal levels of physical parameters and key media components namely temperature, pH, inoculum size, moisture, yeast extract, MgSO4 . 7H20, Tween 80, and corn steep liquor (CSL) were determined. Among the physical parameters, temperature contributed higher influence, and among media components, yeast extract, MgSO4 . 7H20, and Tween 80 played important roles in the conversion of starch to lactic acid. The expected yield of lactic acid under these optimal conditions was 95.80% and the actual yield at optimum conditions was 93.50%.

Bacteriological Techniques↗
PubMed2003

Characterization of Lactobacillus sakei by the type of stereoisomers of lactic acid produced.

Lactobacillus sakei strains were characterized by the shift of the type of stereoisomers of lactic acid produced in the presence of 50 mM sodium acetate in a medium. Of 27 Lactobacillus sakei strains studied, 20 strains showed high levels of DNA-DNA similarity with L. sakei NRIC 1071(T), and were confirmed as L. sakei. The three remaining strains were identified as Lactobacillus curvatus by DNA-DNA similarity, and three other strains were included in the cluster of Lactobacillus plantarum/Lactobacillus pentosus/Lactobacillus paraplantarum and one strain in the cluster of Lactobacillus paracasei on the basis of 16S rRNA gene sequences. Of the 20 L. sakei strains, 19 strains shifted the type of stereoisomers of lactic acid produced from the DL-type to the L-type in the presence of 50 mM sodium acetate. L. curvatus strains and strains included in the cluster of L. plantarum/L. pentosus/L. paraplantarum and in the cluster of L. paracasei did not shift the type of stereoisomers of lactic acid produced. The change of the type of stereoisomers of lactic acid from the DL-type to the L-type in the presence of sodium acetate was concluded to be species-specific for L. sakei and useful for identification of strains in this species.

DNA, Bacterial↗
PubMed1991

Spectrophotometric method for the determination of microquantities of lactic acid in biological material.

A simple spectrophotometric method for the determination of microquantities of lactic acid is described. Lactic acid was quantitatively oxidized to acetaldehyde by cerium(IV) under prescribed conditions. Oxidation was terminated by reducing cerium(IV) to cerium(III) with nitrite. The acetaldehyde was reacted with copper ions and oxalydihydrazide under alkaline conditions to form an intense blue complex which is measured spectrophotometrically. The developed citrate-phosphate-borate-sodium hydroxide alkalizing reagent enhanced color stability by removing cerium from solution and also ensured extremely low blank values. Beer's law was followed over the range 0 to 150 micrograms of lactate. The method was successfully applied to the routine determination of lactate in silage and blood.

Biological Factors↗
PubMed2000

Modeling of direct recovery of lactic acid from whole broths by ion exchange adsorption.

Lactic acid fermentation process with L. casei CRL 686 was performed. The static adsorption isotherm over a strong anionic exchange resin, Amberlite IRA-400 was measured, and the static binding capacity parameters were quantified. Early recovery of lactic acid from this lactate producer from unclarified culture broth was performed in a liquid solid fluidized bed, with the resin as the solid adsorbent, and the dynamic adsorption capacity was calculated. Good agreement was found between static and dynamic binding capacity values. The fluidized bed height was twice the settled bed height and the overall process was controlled by the liquid solid mass transfer. This operation was also simulated by continuously well stirred tanks arranged in series and superficial solid deactivation as in a gas solid catalytic reactor. The deactivation process takes into account liquid channeling and agglomerations of solid induced by the viscosity of the broth and also by the cells during the adsorption. These patterns were also verified by experimental observations, and are in agreement with the results found in the literature. The breakthrough data together with others from previous works were satisfactorily fitted until the 90% dimensionless concentration was reached for both culture broths. The model could be used in future studies on predictions about the liquid solid fluidized bed behavior and other different operating conditions.

Adsorption↗
PubMed2001

Simultaneous determination of D-lactic acid and 3-hydroxybutyric acid in rat plasma using a column-switching HPLC with fluorescent derivatization with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ).

A highly sensitive method for the determination of D-lactic acid and 3-hydroxybutyric acid (3-HB) in rat plasma was developed using high-performance liquid chromatography with octadecylsilica (ODS) connected to a chiral column. At first, (D + L)-lactic acid and 3-HB in the plasma were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on the ODS column and determined fluorimetrically at 547 nm with 491 nm of excitation wavelength. During the separation step on the ODS, the peak fraction of (D + L)-lactate derivative was introduced directly to a phenylcarbamoylated beta-cyclodextrin chiral column by changing the flow of the eluent via a six-port valve. Then, D-lactate derivative was separated enantiomerically from the L-lactate derivative, and the enantiomeric ratio was determined from the chromatogram. Intra- and inter-day accuracy values for the determination of D-lactic acid in 10 microL of rat plasma were 97.8-109.2 and 98.4-109.9%, and those for 3-HB were 99.8-108.4 and 99.8-103.8%, respectively. The intra- and inter-day precision values were within 4.6 and 5.1% for D-lactic acid, and 2.7 and 2.4% for 3-HB, respectively. The detection limits for D-lactic acid and 3-HB were approximately 2.0 and 0.04 microM, respectively (signal-to-noise ratio 3). The proposed method was applied to the plasma of diabetic rats induced by intraperitoneal administration of streptozotocin, and the significant increases of both D-lactic acid and 3-HB concentrations were observed in the diabetic rats as compared to the normal rats.

3-Hydroxybutyric Acid↗
PubMed1993

Characterization and frequency distribution of species of lactic acid bacteria involved in the processing of mawè, a fermented maize dough from Benin.

Lactic acid bacteria involved in the natural fermentation of both home-produced and commercial mawè were investigated during a 72 h fermentation period. Lactobacillus spp. constitute the majority (94%) of the strains of the lactic acid bacteria isolated, among which 89% represent the Betabacterium group. They include L. fermentum (biotype cellobiosus) (41%), L. fermentum or L. reuteri (19%), L. brevis (26%), L. confusus (less than 2%), L. curvatus (less than 1%) and L. buchneri (less than 1%). Other isolated lactic acid bacteria were L. salivarius, Lactococcus lactis, Pediococcus pentosaceus, Pediococcus acidilactici and Leuconostoc mesenteroides. Several species were detected at the early stage of fermentation, but the final stage was dominated by L. fermentum (biotype cellobiosus) and L. fermentum or L. reuteri totalling 90% of the isolated strains. The trend was the same for both home-produced and commercial mawè. No strains of L. plantarum, generally reported as dominating lactic acid bacteria at the final stage of fermentation of most plant foods, were isolated.

Benin↗
PubMed1988

Effects of lactic acid infusions and pH on cerebral blood flow and metabolism.

To determine the effects of lactic acidemia versus lactate on CBF, we infused lactic acid, either buffered with NaOH (L + NaOH) or with added NaCl (L + NaCl), to attain similar osmolalities in 18 piglets. CBF (microsphere technique), pH, blood gases, plasma osmolality, and cerebral arteriovenous differences of O2 content and lactic acid concentrations were measured prior to, at 30 min of a lactic acid infusion, and 15 and 90 min after completion of the infusion. Control arterial pH was comparable between groups (7.50 +/- 0.02 vs. 7.49 +/- 0.02, X +/- SE); during and following L + NaCl and L + NaOH, values were (p less than 0.05) 7.09 +/- 0.03, 7.35 +/- 0.02, and 7.46 +/- 0.02 vs. 7.58 +/- 0.03, 7.61 +/- 0.01, and 7.57 +/- 0.03, respectively. PaCO2 remained unchanged and osmolality rose by 15% in both groups during infusions and persisted throughout the study period. For L + NaCl piglets, CBF (ml/min.100 g) rose from 136 +/- 15 to 198 +/- 26 (p less than 0.05) at 30 min of infusion and remained elevated at 201 +/- 25 and 207 +/- 28 at 15 and 90 min following the infusion, respectively. Similarly, for L + NaOH piglets, CBF rose from 130 +/- 25 to 196 +/- 31 (p less than 0.05) with the infusion and was 174 +/- 17 and 166 +/- 21 at 15 and 90 min afterward, respectively. Although lactic acid infusion increases CBF, the associated metabolic acidemia is not responsible for changes in CBF.

Animals↗
PubMed1997

Lactic acid potentiates bradykinin- and low-pH-induced release of CGRP from rat spinal cord slices.

Previous data from our laboratory have shown that calcitonin gene-related peptide (CGRP) is released into the circulation during pathogenesis of endotoxic, hemorrhagic, and septic shock and appears to mediate in part the vascular problems of shock. Elevations in the levels of bradykinin (BK) and lactic acid and lowering of tissue pH also occur during shock and could be involved in CGRP release. In the present study, we have tested whether lactic acid, alone or in combination with BK or low pH, triggers release of CGRP-like immunoreactivity (CGRP-LI) from sensory nerves, using rat spinal cord slices as a tissue model. Lowering media pH from 7.4 to < or = 6.0 increased the release of CGRP-LI. Lactic acid (5 and 10 mM) by itself elevated CGRP-LI release from a control of 6.89 +/- 0.95 to 57.2 +/- 8.2 and 116 +/- 13 pg/mg protein, respectively. The combination of pH 6.0 and lactic acid (5 or 10 mM) caused more than additive stimulation of CGRP-LI release. BK (50 or 100 microM) elevated CGRP-LI release, which was greatly enhanced by lactic acid (2.5 or 5 mM). The data indicate that lactic acid potentiates BK- and low-pH-induced release of CGRP from sensory nerves in spinal cord. Similar mechanisms may occur at peripheral ends of sensory nerves, contributing to CGRP release during septic shock and other conditions with elevated lactic acid levels (e.g., strenuous exercise and tissue ischemia).

Animals↗
PubMed1996

Migration of press-fit cups fixed with poly-L-lactic acid or titanium screws: a randomized study using radiostereometry.

The early migration of porous acetabular cups was studied in 43 patients with osteoarthrosis. The patients were randomly allocated to additional fixation of the cup with either biodegradable poly-L-lactic acid screws or titanium screws. Radiostereometric evaluation was done during the first 2 years after the procedure in 43 hips (23 with poly-L-lactic acid screws and 20 with titanium screws). At the 2-year follow-up, cups fixed with poly-L-lactic acid screws had migrated significantly more in the proximal-distal (p < 0.05) and medial-lateral (p < 0.05) directions. Cups with titanium screws displayed more pronounced rotations around the longitudinal axis (p < 0.05). Postoperatively, on the lateral view, there was an increased occurrence of radiolucencies at the dome of the cups fixed with poly-L-lactic acid screws (p < 0.05). The clinical result did not differ between the two groups. Inferior implant-bone contact in the poly-L-lactic acid group, local changes of the bone quality, and diminishing support of the poly-L-lactic acid screws caused by their degradation with time could be reasons for the different pattern of migration observed.

Acetabulum↗
PubMed1991

D-lactic acid production as a monitor of the effectiveness of antimicrobial agents.

Most bacteria at an infection site obtain energy by the breakdown of glucose via microaerophilic or anaerobic pathways and in the process yield various end products. In this study, production of D-lactic acid by Staphylococcus aureus and Escherichia coli was correlated with glucose utilization by bacteria during exposure to antibiotics at subinhibitory, inhibitory, and suprainhibitory concentrations. D-Lactic acid production was further correlated with production of a tissue-destroying enzyme, hyaluronidase, by S. aureus. For E. coli, all agents tested showed dose-related bacterial killing, with the most noticeable being with ampicillin, piperacillin, and ciprofloxacin. Imipenem, ciprofloxacin, and chloramphenicol had the most dose-related effects on D-lactic acid production. With few exceptions, hyaluronidase production correlated well with D-lactic acid production in S. aureus. Subinhibitory concentrations of erythromycin and clindamycin effectively decreased accumulation of D-lactic acid and hyaluronidase. Determination of D-lactic acid production may perhaps serve as a means of independently monitoring the effects of antimicrobial agents on bacterial metabolic activity, which is an important aspect of antimicrobial action that remains relatively unexplored.

Anti-Bacterial Agents↗