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Practical implications of lactate and pyruvate metabolism by lactic acid bacteria in food and beverage fermentations.

This article reviews the metabolism of pyruvate and lactate by lactic acid bacteria (LAB) involved in food and beverage fermentations with an emphasis on practical implications. First, the formation of pyruvate and lactate from a range of substrates, including carbohydrates, organic acids and amino acids, is briefly described. The catabolism of pyruvate and lactate by LAB is then reviewed. This is followed by a discussion of lactate degradation and racemisation by LAB from specific fermented foods and beverages. Finally, the impact of environmental factors and metabolic engineering on pyruvate and lactate metabolism by LAB is evaluated with regard to practical significance.

Beverages↗

Separation of cells and proteins from fermentation broth in a shear-enhanced cross-flow ultrafiltration module as the first step in the refinement of lactic acid.

A shear-enhanced, cross-flow ultrafiltration module was used to separate cells and proteins from the fermentation broth. Three (fermented) media were studied: rich medium, rich medium with hydrolytic enzymes added after fermentation, and wheat flour hydrolysate. To find a membrane with as high a flux as possible, but still capable of separating cells and proteins from the lactic acid containing broth, the performance of three hydrophilic membranes of varying cutoffs (10,000, 20,000, and 30,000) and one hydrophobic membrane (cutoff 25,000) was investigated. The proteins produced by the lactic acid bacteria during fermentation and the hydrolytic proteins were retained by the hydrophilic membrane with a cutoff of 20,000, whereas wheat flour proteins were detected in the permeate. In the permeates from the hydrophobic membrane (cutoff 25,000), almost no proteins were detected. The flux of the whole-wheat flour hydrolysate was significantly lower than that of rich medium, for both the hydrophilic and the hydrophobic membranes. The flux was, in all cases, higher for the hydrophilic membrane (12-85 L/[m2 x h], depending on which medium was treated) than for the hydrophobic one (8-45 L/[m2 x h]), even though the nominal cutoffs of the hydrophobic and hydrophilic membranes were almost the same. However, the difference in flux was smaller when the whole-wheat flour hydrolysate was processed (12 vs 8 L/[m2 x h]) than when the rich medium was processed (85 vs 45 L/[m2 x h]). Protein retention was higher for the hydrophobic membrane than for the hydrophilic membrane (cutoff 20,000) owing to blocking of the pores by proteins adsorbed on to the hydrophobic membrane surface.

Journal Article↗

Relations between ion shifting, ATP depletion and lactic acid formation in human red cells during moderate calcium loading using the ionophore A 23187.

Keeping constant cellular magnesium an A 23 187 mediated moderate calcium loading of human red cells causes isoosmotic cell shrinkage, potassium efflux, slight decrease of cellular pH, ATP depletion connected with an increase of AMP, ADP and Pi and enhanced lactic acid formation. The calcium loading and accompanying effects can be abolished by EGTA or by extracellular magnesium, the latter kept more than two orders of magnitude above that of calcium which was 30 micrometer. Inhibition of the (Mg2+ + Ca2+)-dependent ATPase by ruthenium red or lanthanum decreases the calcium stimulated lactic acid formation after a lag phase. However, the ATP depletion proceeds faster and is much more pronounced under these conditions. (Mg+2 + Na+ +K+)-dependent ATPase, hexokinase, phosphofructokinase and cell shrinkage are ruled out, too, as mediators of the ATP depletion. This suggests that an unknown ATP consuming reaction, apparently not being related to the calcium pump, causes the calcium induced ATP depletion.

Adenosine Triphosphate↗

[Metabolism of lactic acid stereoisomers in living organisms].

The present state of one of the sections of dynamic biochemistry concerning lactic acid enantiomers metabolism has been examined. Data on the distribution and activity of L- and D-lactate NAD+ oxidoreductases in various biological objects have been summed up. Probable involvement of prochiral and chiral compounds that stimulated biogenesis in the processes of quasi-metabolism self-organization and chiral purity of biosphere has been supposed.

Animals↗

Survival of lactic acid bacteria in simulated gastrointestinal juice protected by a DNA-based complex gel.

Salmon milt DNA was utilized in the approach to protect functional materials, proteins, microorganisms, drugs, etc., by forming a DNA-based complex gel with gelatin and kappa-carrageenan. The multiplier effect of the combination of DNA, gelatin and kappa-carrageenan was examined by measuring the release of methyl orange from the gel after being incubated in simulated gastric juice. The results indicated that the hydrogel made of DNA, gelatin and kappa-carrageenan can efficiently maintain the functional materials. Furthermore, a complex gel was developed and optimized by applying an emulsion of lactic acid bacteria and cacao oil onto the already prepared hydrogel. The existence of the lactic acid bacteria in the oil emulsion particles was confirmed by scanning electron microscopy. The protection capabilities in the simulated gastrointestinal juice, which had a viability of 1.50 x 10(6) CFU/g gel after 2 h incubation in the simulated gastric juice and 2.53 x 10(7) CFU/g gel after 6 h continued incubation in the simulated intestinal juice, were measured, in contrast to 2.58 x 10(7) CPU/g in the original gel before treatment.

Carrageenan↗

An introduction to rod-shaped lactic-acid bacteria.

The genus Lactobacillus is mainly found in the intestinal tract of healthy humans and animals as well as in fermenting vegetables or plant materials, such as silage. It has a moderate diffusion in meat products and is rarely found in wines and beers. On the other hand, rod-shaped lactic-acid bacteria are largely used in the preparation of a variety of foods and feed products. As starter cultures, they are omnipresent in cheeses and dairy manufacturing. Specific fermentation processes have been developed in order to encourage the growth of the desired species, some of which are fastidious organisms such as L. delbrueckii ssp. bulgaricus, L. helveticus and L. sanfrancisco. In addition, a promising and interesting perspective is the use of rod-shaped lactic-acid bacteria, primarily L. acidophilus, L. reuteri and L. salivarius as probiotic starters to preserve the natural biological equilibrium of the intestinal tract.

Lactobacillus↗

Poly-L-lactic acid: an overview.

In August 2004, the US Food and Drug Administration approved a poly-L-lactic acid (PLLA)-based injectable medical device for restoration and/or correction of the signs of facial fat loss (lipoatrophy) in people with human immunodeficiency virus. As a result, the properties of the PLLA microparticles have received considerable interest from the medical community. Polylactides have a long-standing history of safe use in medical applications, such as pins, plates, screws, intra-bone and soft-tissue implants, and as vectors for sustained release of bioactive compounds. The L-isomer of polylactic acid is a biodegradable, biocompatible, biologically inert, synthetic polymer. Putatively, PLLA microparticles initiate neocollagenesis as a result of a normal foreign-body reaction to their presence. The build-up of collagen over time creates volume at the site of injection, while the PLLA microparticles are metabolized to carbon dioxide and water and expelled through the respiratory system.

Absorbable Implants↗

Influence of sodium nitrite, temperature, and lactic acid bacteria on the growth of Brochothrix thermosphacta under anaerobic conditions.

In commercial vacuum-packed bologna, growth of Brochothrix thermosphacta was limited at 5 degrees C during a 4-week storage period. Factors controlling the growth of this organism were associated in part with certain lactobacilli isolated from the meat. The lactic acid bacteria associated with the decline of B. thermosphacta and overall spoilage of vacuum-packed bologna were isolated and identified. The principal organisms were Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus plantarum, Lactobacillus viridescens, Leuconostoc mesenteroides, and certain streptococci. In associated growth experiments with B. thermosphacta at 5 and 15 degrees C, L. brevis and L. plantarum showed the greatest antagonism towards the Brochothrix species. Studies with changes in incubation temperatures (5 or 15 degrees C) or nitrite levels (0-50 microgram/mL) indicated that these two factors influence the antagonism but were minor compared with the influence of the lactic acid bacteria.

Anaerobiosis↗

Potential of using lactic acid bacteria for therapy and immunomodulation in man.

There is in 1993 no proven medical indication of lactic acid bacteria (LAB) for therapy or immunomodulation in man. However, within the bulk of publications, rigorous trials have now opened rational fields of research on beneficial effects of LAB. These include lactose digestion, cholesterol metabolism, diarrheal disorders, prophylaxis of intestinal or urogenital infections, immunomodulation or even oral vaccination. We try here to analyse these studies, considering LAB as pharmacological agents, and conclude that pursuit of research could be promising for ecological therapy of mucosal diseases, and for development of original and flexible vectors for targeting in the gastrointestinal tract.

Colonic Neoplasms↗

Chryseobacterium shigense sp. nov., a yellow-pigmented, aerobic bacterium isolated from a lactic acid beverage.

A yellow-pigmented bacterium designated strain GUM-Kaji(T) was isolated from a lactic acid beverage. The strain had Gram-negative, non-motile, rod-shaped cells. It was strictly aerobic and chemo-organotrophic and grew at 5-30 degrees C and at pH 5-8. The major components of the non-polar and 3-hydroxy fatty acids were C15:0 iso and 3-OH-C17:0 iso, respectively. Menaquinone MK-6 was detected as the sole quinone. 16S rRNA gene sequence comparisons revealed that strain GUM-Kaji(T) is affiliated to the genus Chryseobacterium, with Chryseobacterium joostei as its phylogenetic neighbour, but there were low levels of similarity (<96%) to any established species of the genus. The G+C content of the genomic DNA was 36.6 mol%. The novel bacterium differed from any known species of Chryseobacterium in terms of a number of phenotypic properties. Thus, the name Chryseobacterium shigense sp. nov. is proposed for this novel bacterium. The type strain is strain GUM-Kaji(T) (=BAMY 1001(T)=NCIMB 14047(T)=DSM 17126(T)).

Aerobiosis↗

The fate of the poly-L-lactic acid interference screw after anterior cruciate ligament reconstruction.

We report the persistence of a poly-L-lactic acid (PLLA) interference screw 2.5 years after anterior cruciate ligament (ACL) reconstruction with Achilles tendon allograft. The arthroscopy was performed because the patient sustained a reinjury of the ACL graft, making ACL revision surgery necessary. At the time of arthroscopy, both PLLA screws were macroscopically still intact but could not be removed in 1 piece. No inflammation could be observed either macroscopically or in the histologic analysis. The biopsy specimen from the femoral insertion of the graft showed parts of the PLLA material surrounded by scar tissue. This case shows that biodegradation of PLLA material in the knee joint causes no irritation and can take several years, even if the material is in contact with the synovial fluid.

Absorbable Implants↗

Surface modification of poly-L-lactic acid (PLLA) membrane by grafting acrylamide: an effective way to improve cytocompatibility for chondrocytes.

Poly-L-lactic acid (PLLA) membranes were photo-oxidized in hydrogen peroxide solution under ultraviolet light (UV) to introduce hydroperoxide groups onto the PLLA membrane surfaces. The photo-oxidized membranes were then immersed in acrylamide (AAm) solution containing Fe2+ to graft polyacrylamide (PAAm) onto the PLLA membrane surfaces. The density of the hydroperoxide groups introduced on the PLLA membrane surfaces varied with the temperature and the photo-oxidization time. The occurrence of grafting was verified by X-ray photoelectron spectroscopy (XPS). The degree of grafting increased with the monomer concentration and the polymerization time. Water contact angle measurements showed that the wettability of the modified PLLA membranes had improved. Chondrocytes proliferated more rapidly and were more spread out on the modified membrane than on the control PLLA membrane, indicating that the PAAm-grafted PLLA membrane has better cytocompatibility for chondrocytes.

Acrylamide↗

Biodegradable poly(lactic acid) and poly(lactide-co-glycolide) microcapsules: problems associated with preparative techniques and release properties.

Poly(lactic acid) [PLA] and its co-polymers with glycolic acid [PLCG] have been known to be biodegradable and histocompatible for the past 20 years. Their physico-chemical and biological properties have been found suitable, in many instances, for sustaining drug release in vivo for days or months. Several dosage forms for parenteral administration have been investigated using these polymers and a microencapsulation technique is chosen frequently for its unique properties. There are a limited number of published papers concerning preparation and characterization of PLA or PLCG microcapsules, possibly because of commercial unavailability and difficulties in the synthesis of reproducible batches of these polymers. However, microcapsules can be made using different traditional and non-traditional techniques containing core materials ranging from biological proteins to synthetic drugs. An attempt is made here to review problems associated with the different microencapsulation techniques using PLA or PLCG. In vivo and in vitro drug release from these microcapsules is also reviewed.

Animals↗

Marinilactibacillus piezotolerans sp. nov., a novel marine lactic acid bacterium isolated from deep sub-seafloor sediment of the Nankai Trough.

A piezotolerant, mesophilic, marine lactic acid bacterium (strain LT20T) was isolated from a deep sub-seafloor sediment core collected at Nankai Trough, off the coast of Japan. Cells were Gram-positive, rod-shaped, non-sporulating and non-motile. The NaCl concentration range for growth was 0-120 g l(-1), with the optimum at 10-20 g l(-1). The temperature range for growth at pH 7.0 was 4-50 degrees C, with the optimum at 37-40 degrees C. The optimum pH for growth was 7.0-8.0. The optimum pressure for growth was 0.1 MPa with tolerance up to 30 MPa. The main cellular phospholipids were phosphatidylglycerols (25 %), diphosphatidylglycerols (34 %) and a group of compounds tentatively identified as ammonium-containing phosphatidylserines (32 %); phosphatidylethanolamines (9 %) were minor components. The fatty acid composition was dominated by side chains of 16 : 0, 14 : 0 and 16 : 1. The G+C content of the genomic DNA was 42 mol%. On the basis of 16S rRNA gene sequence analysis and the secondary structure of the V6 region, this organism was found to belong to the genus Marinilactibacillus and was closely related to Marinilactibacillus psychrotolerans M13-2(T) (99 %), Marinilactibacillus sp. strain MJYP.25.24 (99 %) and Alkalibacterium olivapovliticus strain ww2-SN4C (97 %). Despite the high similarity between their 16S rRNA gene sequences (99 %), the DNA-DNA hybridization levels were less than 20 %. On the basis of physiological and genetic characteristics, it is proposed that this organism be classified as a novel species, Marinilactibacillus piezotolerans sp. nov. The type strain is LT20T (=DSM 16108T=JCM 12337T).

Bacterial Typing Techniques↗

Oral ingestion of lactic-acid bacteria by rats increases lymphocyte proliferation and interferon-gamma production.

The effect of feeding lactic-acid bacteria on indices of functions of lymphocytes obtained from Peyer's patches, peripheral blood and spleen from inbred Wistar-Furth rats were studied. Rats were fed on purified diets supplemented with 350 g milk or yoghurt/kg diet for 4 weeks. At the end of the feeding period, immune cells from the three sites were isolated and proliferation, interferon-gamma production and lymphocyte subset composition were studied. Rats consuming yoghurt had a greater in vitro proliferative response to yoghurt bacteria in the three lymphoid compartments, a greater interferon-gamma production in response to bacteria and concanavalin A in Peyer's patches and spleen, and a greater number of Peyer's patches B lymphocytes than milk-fed rats. Macrophage and T lymphocyte proportions and lymphocyte subset composition in the three sites were unaffected by yoghurt. These results indicate that feeding live bacteria contained in yoghurt may interact with the intestinal immune system, and influence the systemic immune system.

Animals↗

The study of biocompatibility of super high molecular weight poly D,L-lactic acid implant.

OBJECTIVE: To evaluate the biocompatibility of the super high molecular weight poly D,L-lactic acid (SHMW-PDLLA) implant. METHODS: The SHMW-PDLLA plates were implanted into the SD-rats between the masseter and ramus of the mandible. The blood specimens were gained at 3, 6, 9, 12 months after the operation. The proteins, electrolyte, enzyme and other indices were tested by use of Beckman automatic biochemical analysis device. The soft tissue specimens around the SHMW-PDLLA plates were gained at 3, 6, 9, 12 months after the operation and the tissue reaction was observed with the pathological and haematological methods. RESULTS: There were not any abnormal findings in the blood after the SHMW-PDLLA plates implanted in the body of SD-rats. The implanted SHMW-PDLLA plates were degraded gradually in 6 to 12 months after the operation. There was not any abnormal tissue reaction found to the soft tissue around the SHMW-PDLLA plates by histological and pathological observations. CONCLUSIONS: The SHMW-PDLLA implant has a good biocompatibility to SD-rats.

Animals↗

Structure and mechanical properties of poly(D,L-lactic acid)/poly(epsilon -caprolactone) blends.

A series of blends of the biodegradable polymers poly(D,L-lactic acid) and poly( epsilon -caprolactone) were prepared by varying mass fraction across the range of compositions. Tensile testing was performed at room temperature using an extensometer and the elastic modulus was calculated for each blend. The blends were also tested to failure, and the strain-at-failure and yield stress recorded. While the blend has been shown to have a lower critical solution temperature, the mechanical properties were insensitive to the annealing conditions. Scanning electron microscopy was used to characterize the blend microstructure and poor adhesion was observed at the interface between blend components. Differential scanning calorimetry was performed but the results were somewhat variable, indicating this blend may have complex phase behavior that depends sensitively on the method of preparation. However, nuclear magnetic resonance data indicate the two components are phase separated. A percolation model is used to explain the observed mechanical data and the results are consistent with the predictions of the Kerner-Uemura-Takayangi model. The results of these experiments demonstrate the utility of polymer blending in tuning material properties.

Biocompatible Materials↗