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PubMed2004

Peptones from autohydrolysed fish viscera for nisin and pediocin production.

Various peptones obtained from hydrolysed visceral homogenates of four fishery residues showed their suitability for promoting the growth of lactic acid bacteria, micro-organisms with particularly complex requirements regarding peptidic nutrients. The assay of several treatments with two bacterial species, producers of the two main bacteriocins (nisin and pediocin) demostrated that optimum conditions only imply a brief autohydrolysis at natural pH and room temperature, with subsequent steam-flow stabilisation. Later kinetic analysis of the cultures of both bacteria in the best media provided parameters which, for production of both biomass and bacteriocins (the latter behaved in the majority of cases as a mixed metabolite), indicate comparable or superior results to those found in costly commercial media, specifically recommended for culture of lactic acid bacteria.

Animals↗
PubMed2003

Conformational changes of pediocin in an aqueous medium monitored by fourier transform infrared spectroscopy: a biological implication.

Fourier transform infrared (FTIR) spectroscopy was used to investigate the secondary structure of pediocin PA-1 in different aqueous media in relation to its antimicrobial activity. The experiments were performed at pD (pH meter corrected for deuterium isotope effect) 6, 7, and 8 and during a heating-cooling cycle of 20-80 degrees C. At pD 6, (i.e. pediocin's most active form), the FTIR results show that pediocin adopts an unordered structure with a small contribution of beta-turn. After a heating-cooling cycle, thermally-induced changes in pediocin are reversed and its activity is maintained. Increasing the pD to 7 and 8 leads to a more ordered secondary structure. For these two pD values, an increase in temperature induces an irreversible aggregation of protein as revealed by the amide I' band. The analysis of the Tyr region provides more insight into the aggregation process. In fact, it appears to be a two-step process, involving first the C (carboxy)-terminus of pediocin and then the N (amino)-terminus. This study reveals two major points: (1) the preservation of pediocin flexibility is essential for maintaining its activity; and (2) the aggregation of its C-terminus is sufficient to induce a loss of activity, suggesting that this region plays an important role in the activity of pediocin.

Anti-Bacterial Agents↗
PubMed2000

Differentiation of Tetragenococcus populations occurring in products and manufacturing processes of puffer fish ovaries fermented with rice-bran.

Tetragenococcus strains isolated from the manufacturing process of Japanese puffer fish ovaries fermented with rice-bran were characterized and differentiated phenotypically and genotypically. A total of 413 Tetragenococcus isolates were evaluated. On the basis of five representative substrates, the isolates were grouped into seven groups. An RFLP (restriction fragment length polymorphisms) analysis of the 16S rRNA gene of representative strains of major groups revealed that they could be grouped into two groups: one was identified as the most prominent halophilic lactic acid coccus, Tetragenococcus halophilus, and the other as T. muriaticus, which has recently been added to the genus Tetragenococcus as a new species. Physiologically, the major differences between the two groups were found in the ability to grow in medium not supplemented with NaCl and the fermentation of L-arabinose, sucrose and D-mannitol, and several other carbohydrates.

Animals↗
PubMed2003

Bioprotectives and probiotics for dry sausages.

The microbial stability of dry sausages is determined by the combination and timing of different factors referred to as the hurdle-concept. However, the hurdles present in dry sausage are not sufficient to prevent the survival of Listeria monocytogenes or enterohemorrhagic Escherichia coli O157:H7. Recently bioprotective lactic acid bacteria, which in addition to the production of antimicrobial lactic acid, have been found to contribute to the safety of the dry sausage by producing antimicrobial peptide, i.e. bacteriocins and other low-molecular-mass compounds. Furthermore, the possibilities to use probotics in dry sausage manufacturing process has been addressed recently. As one possible mode of action for probiotics is the production of antimicrobial compounds, lactic acid bacteria may act as both probiotic and bioprotective culture as well as fermenting agent in meat product, such as dry sausage.

Consumer Product Safety↗
PubMed1997

Purification of bacteriocins of lactic acid bacteria: problems and pointers.

Bacteriocins of lactic acid bacteria have been widely studied in recent years. However, there are relatively few studies that describes their biochemical structure. This study may be due to the many challenges associated with the purification of these antimicrobial peptides. This review focuses on the purification procedures used with bacteriocins of lactic acid bacteria and conveys some of the problems associated with this process as well as some of the lessons learned. An improvement in the efficiency of the purification process should contribute significantly to research at the understanding of the biochemical nature of bacteriocins.

Bacteriocins↗
PubMed1997

A computerised system for the identification of lactic acid bacteria.

A generic computerised system for the identification of bacteria was developed. The system is equipped with a key to the identification of lactic acid bacteria. The identification is carried out in two steps. The first step distinguishes groups of bacteria by following a decision tree with general identification tests. The second step in the identification is the distinction of species within a group on the basis of biochemical fermentation patterns. During grouping, probabilities for test failure are used. These probabilities can be used for assessing the quality of a given test answer. The probabilities are also used to select the most probable test answer in cases where the test result is inconclusive. The probabilities of test failure were determined by a group of experts and a group of potential users of the identification system. During species identification, similarity indices are calculated for all bacteria in a group. The described identification system has the ability to "learn" from different sessions in the species identification step, improving both identification speed and accuracy. Because of the versatile way in which the system is set up, it can very easily be expanded with identification keys to other organisms.

Carbohydrate Metabolism↗
PubMed2003

Development of specific fluorescent oligonucleotide probes for in situ identification of wine lactic acid bacteria.

A rapid method for the identification of lactic acid bacteria (LAB) from wine has been developed. This method is based on fluorescence in situ hybridisation (FISH), using fluorescent oligonucleotide probes, homologous to 16S rDNA of those species of LAB commonly found in wines. The protocol for the specific detection of these bacteria was established through the hybridisation of 36 reference strains. The specificity of the probes was evaluated by using pure cultures. Probes were used to identify species in different wines, making it evident that direct identification and quantification from natural samples without culturing is also possible. The results show that FISH is a promising technique for the rapid identification of LAB, allowing positive identification in a few hours (4-16 h).

Base Sequence↗
PubMed1976

Acetaldehyde: an intermediate in the formation of ethanol from glucose by lactic acid bacteria.

Group N streptococci formed acetaldehyde and ethanol from glucose. As the enzymes aldehyde dehydrogenase, phosphotransacetylase and acetate kinase were present this would enable these organisms to reduce acetyl-CoA to acetaldehyde and convert acetyl-CoA to acetyl phosphate and acetate. A pentose phosphate pathway which converted ribose-5-phosphate to glyceraldehyde-3-phosphate was also present. Acetaldehyde could not be formed via the hexose monophosphate shunt or by direct decarboxylation of pyruvate, as the enzymes phosphoketolase and alpha-carboxylase were absent. Phosphoketolase activity was induced in Streptococcus lactis subsp. diacetylactis after growth on D-xylose. Group N streptococci also contained an NAD-dependent alcohol dehydrogenase which reduced acetaldehyde to ethanol while both NAD- and NADP-dependent alcohol dehydrogenase activities were found in Leuconostoc cremoris.

Acetaldehyde↗
PubMed1976

Formation of acetaldehyde from threonine by lactic acid bacteria.

Group N streptococci were found to cleave threonine to form acetaldehyde and glycine. Threonine aldolase, the enzyme catalysing this reaction, was found in all strains except Streptococcus cremoris Z8, an organism which had been shown previously to have a nutritional requirement for glycine. The enzyme was strongly inhibited by glycine and cysteine. The inhibition showed characteristics of allosteric inhibition and was pH-dependent. Inhibition by glycine, but not by cysteine, was highly specific. Analogues and derivatives of cysteine which contained a thiol group and a free amino group inhibited the activity of threonine aldolase. The presence of a carboxyl group was not necessary for inhibition. The cleavage of threonine by whole-cell suspensions was stimulated by either an energy source to aid transport or by rendering the cells permeable to substrate with oleate. Threonine did not appear to be degraded by enzymes other than threonine aldolase, as threonine dehydratase activity was low and NAD- and NADP-dependent threonine dehydrogenases were absent.

Acetaldehyde↗
PubMed2001

Conformational changes in pediocin AcH upon vesicle binding and approximation of the membrane-bound structure in detergent micelles.

Pediocin AcH is a 44-residue antimicrobial peptide with bactericidal potency against Gram-positive bacteria such as Listeria. It belongs to a family of bacteriocins that, when membrane-associated, is predicted to contain beta-sheet and alpha-helical regions. All bacteriocins in this family have a conserved N-terminal disulfide bond. An additional C-terminal disulfide bond in pediocin AcH is thought to confer enhanced potency and broader specificity range against sensitive bacteria. The C-terminal disulfide bond may also affect the conformation of the C-terminus. The secondary structures of pediocin AcH in aqueous solution and vesicles from susceptible cells, as well as the ability of trifluoroethanol (TFE) and detergent systems to induce secondary structures like those induced in vesicles, were studied by circular dichroism (CD) spectroscopy. Like related peptides, pediocin AcH was highly unordered in aqueous solution, 56%. However, it also contained 20% beta-strand and 15% beta-turn structures. Upon complete binding to vesicles, 32% alpha-helical structure formed, the unordered structure decreased to 32%, and the beta-strand and beta-turn structures remained largely unchanged. Thus, a betaalpha domain structure formed in vesicles. The helical structure likely forces the C-terminal tail to loop back on the helix so that the C24-C44 disulfide bond can form. Detergent micelles were superior to TFE in their ability to induce secondary structural fractions in pediocin AcH comparable to those observed in vesicles. This demonstrates the importance of a hydrocarbon-water interface to pediocin AcH structure induction and suggests that it is preferable to use detergent micelles as solvents in NMR studies of pediocin AcH structure.

Amino Acid Sequence↗
PubMed2005

Uracil as an index of lactic acid bacteria contamination of tomato products.

The aim of this research was to evaluate the suitability of uracil as an hygienic quality index of tomato products. Whereas uridine was naturally present throughout tomato fruits' ripening, uracil appeared only after microbial contamination. In tomato pulp inoculated with nine different microbial strains, all five lactic acid bacteria (LAB) studied released relevant quantities of uracil (150-1040 mg/kg of dm), with a correlated partial or total decrease of uridine. Uracil production by yeasts and molds was very low or nonexistent; the starting uridine concentration (approximately 960 mg/kg of dm) remained constant or increased. Uracil thermostability was also verified. Twenty-six samples of tomato paste (30 degrees Brix) were collected from bag-in-drums produced in an industrial processing plant, some with evident swelling symptoms. All of the samples with high microbial count presented uracil. Uracil was also present in samples with microbial contamination under the detection limit and Howard mold count below legislation limits, implying the reprocessing, at least partial, of altered tomato product. The results indicate that uracil presence in tomato products is an index of LAB contamination that has occurred before heat treatment.

Drug Stability↗
PubMed2005

Biogenic amines in wines from three Spanish regions.

One hundred and sixty-three wines from La Rioja, Utiel-Requena, and Tarragona were analyzed to determine if there were any differences in the concentrations of six biogenic amines that are found in these three regions. The influence of grape variety, type of vinification, wine pH, malolactic fermentation, and storage in bottle on biogenic amine concentrations was studied. Results show important differences in putrescine and histamine concentrations among regions, varieties of grape, and type of wine; differences were less appreciable for the remaining biogenic amines studied. Low pH prevented biogenic amine formation. Malolactic fermentation and short storage periods in bottle (3-6 months) showed increases in histamine concentration, whereas longer periods of storage led to a general decrease in histamine. Several strains of lactic acid bacteria were isolated in this work, and their ability to form biogenic amines was assayed in synthetic media, grape must, and wine. Grape varieties, different types of winemaking, pH, and lactic acid bacteria may be responsible for the differences observed in the biogenic amine concentrations of the wines analyzed.

Biogenic Amines↗