Aims: To determine structure–function relationships of antibacterial phenolic acids and their metabolites produced by lactic acid bacteria (LAB). Methods and Results: Minimum inhibitory ...concentrations (MICs) of 6 hydroxybenzoic and 6 hydroxycinnamic acids were determined with Lactobacillus plantarum, Lactobacillus hammesii, Escherichia coli and Bacillus subtilis as indicator strains. The antibacterial activity of phenolic acids increased at lower pH. A decreasing number of hydroxyl groups enhanced the activity of hydroxybenzoic acids, but had minor effects on hydroxycinnamic acids. Substitution of hydroxyl groups with methoxy groups increased the activity of hydroxybenzoic, but not of hydroxycinnamic, acid. Metabolism of chlorogenic, caffeic, p‐coumaric, ferulic, protocatechuic or p‐hydroxybenzoic acids by L. plantarum, L. hammesii, Lactobacillus fermentum and Lactobacillus reuteri was analysed by LC‐DAD‐MS. Furthermore, MICs of substrates and metabolites were compared. Decarboxylated and/or reduced metabolites of phenolic acids had a lower activity than the substrates. Strain‐specific metabolism of phenolic acids generally corresponded to resistance. Conclusions: The influence of lipophilicity on the antibacterial activity of hydroxybenzoic acids is stronger than that of hydroxycinnamic acids. Metabolism of phenolic acids by LAB detoxifies phenolic acids. Significance and Impact of the Study: Results allow the targeted selection of plant extracts for food preservation, and selection of starter cultures for fermented products.
Aims: Isomalto-oligosaccharides (IMO) with α(1→6) and α(1→4) glucosidic linkages are produced by enzymatic conversion of starch. IMO are only partially digestible but data on their influence on ...intestinal microbiota are limited. It was the aim of this study to investigate the effect of IMO diet on intestinal microbiota and short-chain fatty acids production (SCFA) in rats. Methods and results: Three groups of F344 rats, each consisting of six animals, were fed IMO, inulin or a control diets for six weeks. A qualitative assessment of the intestinal microbiota was achieved by PCR-denaturing gradient gel electrophoresis (DGGE). Major bacterial taxa were quantified by quantitative PCR (qPCR), and SCFA were measured using gas chromatography. Quantitative PCR demonstrated that lactobacilli were one of the dominant bacterial taxa in faecal samples from rats. IMO increased the number of lactobacilli and the total number of intestinal bacteria in rats fed IMO compared with animals receiving control and inulin diets. Furthermore, PCR-DGGE with lactobacilli-specific primers showed an altered biodiversity of lactobacilli in rats fed IMO compared with control diet. Conclusions: IMO selectively stimulates lactobacilli and increases their diversity in rats. Significance and impact of study: Isomalto-oligosaccharides specifically stimulate growth of intestinal lactobacilli in a rat model system.
This study determined exopolysaccharide (EPS) production by Weissella cibaria MG1 in sourdoughs prepared from gluten-free flours (buckwheat, oat, quinoa and teff), as well as wheat flour. Sourdoughs ...(SD) were fermented without sucrose, or by replacing 10% flour with sucrose to support EPS production. The amount of EPS depended on the substrate: high amounts of EPS corresponding to low amounts of oligosaccharides were found in buckwheat (4.2 g EPS/kg SD) and quinoa sourdoughs (3.2 g EPS/kg SD); in contrast, no EPS but panose-series oligosaccharides (PSO) were detected in wheat sourdoughs. Organic acid production, carbohydrates and rheological changes during fermentation were compared to the EPS negative control without added sucrose. Corresponding to the higher mineral content of the flours, sourdoughs from quinoa, teff and buckwheat had higher buffering capacity than wheat. Fermentable carbohydrates in buckwheat, teff and quinoa flours promoted W. cibaria growth; indicating why W. cibaria failed to grow in oat sourdoughs. Endogenous proteolytic activity was highest in quinoa flour; α-amylase activity was highest in wheat and teff flours. Protein degradation during fermentation was most extensive in quinoa and teff SD reducing protein peaks 18-29, 30-41 and 43–55 kDa extensively. Rheological analyses revealed decreased dough strength (AF) after fermentation, especially in sucrose-supplemented buckwheat sourdoughs correlating with amounts of EPS. High EPS production correlated with high protein, fermentable sugars (glucose, maltose, fructose), and mineral contents in quinoa flour. In conclusion, W. cibaria MG1 is a suitable starter culture for sourdough fermentation of buckwheat, quinoa and teff flour.
Commercial isomalto‐oligosaccharides (IMO) are functional food ingredients. They are composed of α(1→6)‐ and α(1→4)‐linked oligosaccharides. IMO are partially indigestible, and dietary IMO stimulate ...beneficial members of intestinal microbiota, including lactobacilli and bifidobacteria. However, data on IMO metabolism by lactobacilli are not available. It was the aim of this study to identify metabolic pathways of IMO metabolism in lactobacilli. This study focused on the host‐adapted species Lactobacillus reuteri. Metabolism of bifidobacteria was analysed for comparison. Commercial IMO contained IMO with a degree of polymerization (DP) of up to four and panose‐series oligosaccharides (POS) with a DP of up to 5. Lactobacilli metabolized isomaltose preferentially over oligosaccharides with higher DP. Bifidobacteria preferentially metabolized oligosaccharides with higher DP and accumulated glucose. Metabolism of IMO and POS by L. reuteri was attributed to α(1→6)‐specific glucanase DexB and maltose phosphorylase. Contribution of maltose phosphorylase was verified by quantification of IMO and POS phosphorolysis in crude cellular extracts of L. reuteri 100‐23. In conclusion, metabolism of IMO by lactobacilli is limited to short‐chain oligosaccharides, while bifidobacteria preferentially metabolize oligosaccharides with higher DP. The functionality of commercial IMO can thus be modified by degree of polymerization.
Significance and Impact of the Study
Isomalto‐oligosaccharides (IMO) are applied as functional food ingredients, but the composition and biological functionality of current commercial products are poorly documented. This study is the first to analyse IMO metabolism by Lactobacillus reuteri. Bifidobacteria were used for comparison. Commercial IMO contained IMO with degree of polymerization (DP) of up to four and panose‐series oligosaccharides with DP of up to 5. L. reuteri preferentially metabolized short‐chain oligosaccharides, whereas bifidobacteria preferentially metabolized higher oligosaccharides. Results of this study allow the modification of the biological and technological functionality of commercial IMO by adjustment of the degree of polymerization and will thus facilitate the application development for IMO.
Significance and Impact of the Study: Isomalto‐oligosaccharides (IMO) are applied as functional food ingredients, but the composition and biological functionality of current commercial products are poorly documented. This study is the first to analyse IMO metabolism by Lactobacillus reuteri. Bifidobacteria were used for comparison. Commercial IMO contained IMO with degree of polymerization (DP) of up to four and panose‐series oligosaccharides with DP of up to 5. L. reuteri preferentially metabolized short‐chain oligosaccharides, whereas bifidobacteria preferentially metabolized higher oligosaccharides. Results of this study allow the modification of the biological and technological functionality of commercial IMO by adjustment of the degree of polymerization and will thus facilitate the application development for IMO.
Aims: This study aimed to determine the survival of Escherichia coli strains during steam and lactic acid decontamination interventions currently used by the beef-processing industry, and to ...determine their heat resistance. Methods and Results: Strains were grouped into cocktails of five strains each differing in their RAPD patterns for subsequent identification. Steam and lactic acid treatments on meat reduced cell counts of E. coli strain cocktails by 90-99%. The 20 slaughter plant isolates exhibited only minor variation in their resistance to steam and lactic acid treatments but were more resistant than reference strains (three strains) or isolates from live cattle (seven strains). D₆₀ values of strains from live cattle, and reference strains ranged from 0·1 to 0·5 min, in keeping with literature data. However, D₆₀ values of current slaughter plant isolates ranged between 15 for E. coli DM18.3 and 71 min AW 1.7. Cell counts of E. coli AW 1.7 were reduced by <5 log₁₀ CFU g⁻¹ in ground beef patties cooked to an internal temperature of 71°C. Conclusions: Strains of E. coli that survive cooking of ground beef to the recommended internal temperature of 71°C can be isolated from beef-processing facilities. Significance and Impact of the Study: Pathogen interventions in current commercial beef slaughter may select for extremely heat-resistant strains of E. coli.
Aims
To study the antifungal effects of the potato secondary metabolites α‐solanine, α‐chaconine, solanidine and caffeic acid, alone or combined.
Methods and Results
Resistance to glycoalkaloids ...varied among the fungal species tested, as derived from minimum inhibitory concentrations assays. Synergistic antifungal activity between glycoalkaloids and phenolic compounds was found. Changes in the fluidity of fungal membranes caused by potato secondary plant metabolites were determined by calculation of the generalized polarization values. The results partially explained the synergistic effect between caffeic acid and α‐chaconine and supported findings on membrane disruption mechanisms from previous studies on artificial membranes. LC/MS analysis was used to determine variability and relative amounts of sterols in the different fungal species. Results suggested that the sterol pattern of fungi is related to their resistance to potato glycoalkaloids and to their taxonomy.
Conclusion
Fungal resistance to α‐chaconine and possibly other glycoalkaloids is species dependent. α‐Chaconine and caffeic acid show synergistic antifungal activity. The taxonomic classification and the sterol pattern play a role in fungal resistance to glycoalkaloids.
Significance and Impact of the Study
Results improve the understanding of the antifungal mode of action of potato secondary metabolites, which is essential for their potential utilization as antifungal agents in nonfood systems.
Reutericyclin is an inhibitory compound produced by sourdough isolates of Lactobacillus reuteri that is structurally but not functionally related to naturally occurring tetramic acids. It is ...bacteriostatic or bactericidal to gram-positive bacteria based on its activity as a proton-ionophore, and a broad range of food-related spoilage organisms and pathogens is inhibited by reutericyclin. Gram-negative bacteria are resistant to reutericyclin because of the barrier properties of their outer membrane, and resistance of beer-spoiling lactobacilli towards hop bitter acids provides cross-protection to reutericyclin. Remarkably, reutericyclin-producing strains were shown to persist for a period of 10 years in an industrial sourdough fermentation, and reutericyclin was shown to be produced in concentrations active against competitors during growth of L. reuteri in sourdough. Based on the known properties of reutericyclin and L. reuteri, reutericyclin-producing strains may have applications in the biopreservation of foods. Furthermore, these strains were shown to colonize reconstituted lactobacilli-free mice at high levels. Therefore, they could serve as a suitable model system to evaluate a possible impact of antimicrobial compounds on the intestinal microflora of humans and animals.
It was the aim of our work to investigate glutamine deamidation by lactic acid bacteria isolated from cereal fermentations and to elucidate the ecological and technological relevance in baking of the ...conversion of glutamine to glutamate. Lactobacillus sanfranciscensis and Lact. reuteri were found to display glutaminase activity. The addition of glutamine to modified Man, Rogosa and Sharp medium increased the cell yields of Lact. sanfranciscensis, as well as the production of lactic and acetic acid. The final pH; however, was increased in the glutamine-containing medium. The addition of 47 mmol kg⁻¹ glutamate to chemically acidified doughs significantly changed the bread flavour. In sourdoughs with enhanced proteolytic activity, strain-dependent production of 27-120 mmol glutamate per kilogram sourdough was observed. Lactobacillus sanfranciscensis and Lact. reuteri converted glutamine into glutamate; this conversion improves the acid tolerance of lactobacilli and significantly influences wheat bread flavour. This paper illustrates the complex interaction of sourdough-lactobacilli with their environment: the flour provides substrates for metabolic activities that enable the lactobacilli to reach higher cell counts, and the produced metabolite may be one of the reasons why the flavour of fermented breads is different to the flavour of chemically acidified breads.
EPS formed by lactobacilli in situ during sourdough fermentation may replace hydrocolloids currently used as texturizing, antistaling, or prebiotic additives in bread production. In this study, a ...screening of >100 strains of cereal-associated and intestinal lactic acid bacteria was performed for the production of exopolysaccharides (EPS) from sucrose. Fifteen strains produced fructan, and four strains produced glucan. It was remarkable that formation of glucan and fructan was most frequently found in intestinal isolates and strains of the species Lactobacillus reuteri, Lactobacillus pontis, and Lactobacillus frumenti from type II sourdoughs. By the use of PCR primers derived from conserved amino acid sequences of bacterial levansucrase genes, it was shown that 6 of the 15 fructan-producing lactobacilli and none of 20 glucan producers or EPS-negative strains carried a levansucrase gene. In sourdough fermentations, it was determined whether those strains producing EPS in MRS medium modified as described by Stolz et al. (37) and containing 100 g of sucrose liter-1 as the sole source of carbon also produce the same EPS from sucrose during sourdough fermentation in the presence of 12% sucrose. For all six EPS-producing strains evaluated in sourdough fermentations, in situ production of EPS at levels ranging from 0.5 to 2 g/kg of flour was demonstrated. Production of EPS from sucrose is a metabolic activity that is widespread among sourdough lactic acid bacteria. Thus, the use of these organisms in bread production may allow the replacement of additives.
Aims
This study evaluated the aerobic and respiratory metabolism in Lactobacillus reuteri and Lactobacillus spicheri, two heterofermentative species used in sourdough fermentation.
Methods and ...Results
In silico genome analysis, production of metabolites and gene expression of pyruvate oxidase, pyruvate dehydrogenase and cytochrome oxidase were assessed in anaerobic and aerobic cultures of Lact. reuteri and Lact. spicheri. Respiring homofermentative Lactobacillus casei N87 and Lact. rhamnosus N132 were used for comparison.
Aerobiosis and respiration increased the biomass production of heterofermentative strains compared to anaerobic cultivation. Respiration led to acetoin production by Lact. rhamnosus and Lact. casei, but not in heterofermentative strains, in which lactate and acetate were the major end‐products. Lactobacillus spicheri LP38 showed the highest oxygen uptake. Pyruvate oxidase, respiratory cytochromes, NADH oxidase and NADH peroxidase were present in the genome of Lact. spicheri LP38. Both Lact. spicheri LP38 and Lact. rhamnosus N132 overexpressed pox in aerobic cultures, while cydA was up‐regulated only when haeme was supplied; pdh was repressed during aerobic growth.
Conclusions
Aerobic and respiratory growth provided physiological and metabolic advantages also in heterofermentative lactobacilli.
Significance and Impact of the Study
The exploitation of oxygen‐tolerant phenotypes of Lact. spicheri may be useful for the development of improved starter cultures.
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