Teichoic acids possessing phosphate-sugar linkages in strains of Lactobacillus plantarum.
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ABSTRACT:
SUBMITTER: Adams JB
PROVIDER: S-EPMC1184619 | biostudies-other | 1969 Jun
REPOSITORIES: biostudies-other
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Project description:The microbial environment influences animal physiology. However, the underlying molecular mechanisms of such functional interactions are largely undefined. Previously, we showed that during chronic undernutrition, strains of Lactobacillus plantarum, a major commensal partner of Drosophila, promote host juvenile growth and maturation partly through enhanced expression of intestinal peptidases. By screening a transposon insertion library of Lactobacillus plantarum in gnotobiotic Drosophila larvae, we identify a bacterial cell-wall-modifying machinery encoded by the pbpX2-dlt operon that is critical to enhance host digestive capabilities and promote animal growth and maturation. Deletion of this operon leads to bacterial cell wall alteration with a complete loss of D-alanylation of teichoic acids. We show that L. plantarum cell walls bearing D-alanylated teichoic acids are directly sensed by Drosophila enterocytes to ensure optimal intestinal peptidase expression and activity, juvenile growth and maturation during chronic undernutrition. We thus conclude that besides peptidoglycan, teichoic acid modifications participate in the host-commensal bacteria molecular dialogue occurring in the intestine.
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Project description:BACKGROUND: Specific strains of Lactobacillus plantarum are marketed as health-promoting probiotics. The role and interplay of cell-wall compounds like wall- and lipo-teichoic acids (WTA and LTA) in bacterial physiology and probiotic-host interactions remain obscure. L. plantarum WCFS1 harbors the genetic potential to switch WTA backbone alditol, providing an opportunity to study the impact of WTA backbone modifications in an isogenic background. RESULTS: Through genome mining and mutagenesis we constructed derivatives that synthesize alternative WTA variants. The mutants were shown to completely lack WTA, or produce WTA and LTA that lack D-Ala substitution, or ribitol-backbone WTA instead of the wild-type glycerol-containing backbone. DNA micro-array experiments established that the tarIJKL gene cluster is required for the biosynthesis of this alternative WTA backbone, and suggest ribose and arabinose are precursors thereof. Increased tarIJKL expression was not observed in any of our previously performed DNA microarray experiments, nor in qRT-PCR analyses of L. plantarum grown on various carbon sources, leaving the natural conditions leading to WTA backbone alditol switching, if any, to be identified. Human embryonic kidney NF-?B reporter cells expressing Toll like receptor (TLR)-2/6 were exposed to purified WTAs and/or the TA mutants, indicating that WTA is not directly involved in TLR-2/6 signaling, but attenuates this signaling in a backbone independent manner, likely by affecting the release and exposure of immunomodulatory compounds such as LTA. Moreover, human dendritic cells did not secrete any cytokines when purified WTAs were applied, whereas they secreted drastically decreased levels of the pro-inflammatory cytokines IL-12p70 and TNF-? after stimulation with the WTA mutants as compared to the wild-type. CONCLUSIONS: The study presented here correlates structural differences in WTA to their functional characteristics, thereby providing important information aiding to improve our understanding of molecular host-microbe interactions and probiotic functionality.
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Project description:1. The identities of the component glycerol glucosides of the wall teichoic acids of Lactobacillus plantarum N.I.R.D. C106 have been confirmed by methylation analysis. These glucosides are alpha-d-glucopyranosyl-(1-->1)-l-glycerol, alpha-d-glucopyranosyl-(1-->2)-alpha-d-glucopyranosyl-(1-->1)-l-glycerol and alpha-d-glucopyranosyl-(1-->3)-alpha-d-glucopyranosyl-(1-->1)-l-glycerol. 2. These units are connected by phosphodiester groups attached to the 3(l)-hydroxyl group of glycerol and the 6-hydroxyl group of the non-reducing terminal glucose residues in the adjacent unit. 3. Concanavalin A forms a precipitate with the teichoic acid and the material so precipitated contains only the alpha-d-glucopyranosyl-(1-->2)-alpha-d-glucopyranosyl -(1-->1)-l-glycerol component. This unit is therefore present in a homogeneous polymer so that the teichoic acid is a mixture of this and of other possibly homogeneous chains containing the other two components.
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Project description:Naturally fermented pickles harbour many lactic acid bacteria (LAB). Forty-three LAB strains with conjugated linoleic acid (CLA)-producing ability were isolated from three naturally fermented pickle brines. Of these isolates, lp15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability (26.1% conversion) at 48 h in de Man Rogosa Sharpe (MRS) broth in the presence of 100 µg/ml of linoleic acid (LA). Compared to other strains, L. plantarum strain lp15 showed the highest tolerance upon increased levels of LA in the medium, i.e., up to 600 µg/ml. This strain converted about 25% of LA into CLA isomers [predominantly cis-9, trans-11 CLA (9-CLA) and trans-10, cis-12 CLA (10-CLA)], of which 75% was 9-CLA. Interestingly, though the conversion rate of LA into CLA by lp15 remained stable between 100 to 600 µg/ml LA levels in the medium, it dropped sharply at 1000 µg/ml. Taken together, the lp15 strain displayed relatively high LA tolerance with higher conversion rate, which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.
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Project description:We report here the genome sequences of four Lactobacillus plantarum strains which vary in surface hydrophobicity. Bioinformatic analysis, using additional genomes of Lactobacillus plantarum strains, revealed a possible correlation between the cell wall teichoic acid-type and cell surface hydrophobicity and provide the basis for consecutive analyses.
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Project description:A comparative study of two strains of Lactobacillus plantarum (REB1 and MLBPL1) grown in commercial medium (MRS broth), cucumber juice, and liquid pig feed was performed to explore changes to the metabolic pathways of these bacteria, using a proteomics approach (two-dimensional electrophoresis and liquid chromatography-tandem mass spectrometry) combined with analyses of fermentable sugars and fermentation end products. The protein expression showed that even with an excess of glucose in all media, both strains could metabolize different carbohydrates simultaneously and that hexoses could also be used via a phosphoketolase pathway with preferential expression in liquid feed. Sugar analyses showed that the fermentation of sugars was homolactic for all media, with some heterolactic activity in liquid feed, as shown by the production of acetate. Cucumber juice (the medium with the highest glucose content) showed the lowest hexose consumption (10%), followed by liquid feed (33%) and MRS broth (50%). However, bacterial growth was significantly higher in cucumber juice and liquid feed than in MRS broth. This discrepancy was due to the growth benefit obtained from the utilization of the malate present in cucumber juice and liquid feed. Despite different growth conditions, the synthesis of essential cellular components and the stress response of the bacteria were unaffected. This study has improved our understanding of the mechanisms involved in the growth performance of an appropriate lactic acid bacterium strain to be used for food and feed fermentation, information that is of crucial importance to obtain a high-quality fermented product.
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2017-07-12 | GSE101169 | GEO
Project description:We report the identification and chemical characterization of four antifungal substances, 3-(R)-hydroxydecanoic acid, 3-hydroxy-5-cis-dodecenoic acid, 3-(R)-hydroxydodecanoic acid and 3-(R)-hydroxytetradecanoic acid, from Lactobacillus plantarum MiLAB 14. The concentrations of the 3-hydroxy fatty acids in the supernatant followed the bacterial growth. Racemic mixtures of the saturated 3-hydroxy fatty acids showed antifungal activity against different molds and yeasts with MICs between 10 and 100 micrograms ml-1.
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Project description:To investigate the effects of Lactobacillus plantarum (L. plantarum) CAI6 and L. plantarum SC4 on hyperlipidemic mice.Male Kunming mice were fed a high-cholesterol diet for 28 d to construct hyperlipidemic models. Hyperlipidemic mice and normal mice were assigned to 3 groups which were separately treated with L. plantarum CAI6, L. plantarum SC4, and physiological saline through oral gavage for 28 d. Total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels were measured by commercially available enzyme kits. FACS Calibur flow cytometry was used to examine hepatic and renal nuclear factor-erythroid 2-related factor 2 (Nrf2) expression. The morphology of livers was checked by hematoxylin and eosin staining and optical microscope observation.Compared with normal mice, hyperlipidemic mice possessed significantly higher TC (3.50 ± 0.43 vs 2.89 ± 0.36, P < 0.01), TG (1.76 ± 0.07 vs 1.10 ± 0.16, P < 0.01), and LDL-C (1.72 ± 0.20 vs 0.82 ± 0.10, P< 0.01) levels, resulting in an increase of atherogenic index (AI) (2.34 ± 1.60 vs 0.93 ± 0.55, P < 0.05) and LDL-C/HDL-C ratio (1.43 ± 0.12 vs 0.51 ± 0.16, P < 0.05). After treatment with L. plantarum CAI6/L. plantarum SC4, TG (1.43 ± 0.27/1.54 ± 0.10 vs 1.76 ± 0.07, P < 0.01/P < 0.05) and LDL-C (1.42 ± 0.07/1.47 ± 0.12 vs 1.72 ± 0.20, P < 0.01/P < 0.01) in hyperlipidemic mice significantly decreased. In addition, TC, HDL-C, AI, and LDL-C/HDL-C ratio were all positively changed. Meanwhile, the treatment markedly alleviated hepatic steatosis and significantly stimulated Nrf2 expression (73.79 ± 0.80/72.96 ± 1.22 vs 54.94 ± 1.84, P < 0.01/P < 0.01) in hepatocytes of hyperlipidemic mice.L. plantarum CAI6 and L. plantarum SC4 may protect against cardiovascular disease by lipid metabolism regulation and Nrf2-induced antioxidative defense in hyperlipidemic mice.
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Project description:The objective of this research was the identification and characterization of lactic acid bacteria (LAB) isolated from Peruvian Amazonian fruits. Thirty-seven isolates were obtained from diverse Amazonian fruits. Molecular characterization of the isolates was performed by ARDRA, 16S-23S ITS RFLP and rep-PCR using GTG5 primers. Identification was carried out by sequencing the 16S rDNA gene. Phenotypic characterization included nutritional, physiological and antimicrobial resistance tests. Molecular characterization by Amplified Ribosomal DNA Restriction Analysis (ARDRA) and 16S-23S ITS RFLP resulted in four restriction profiles while GTG5 analysis showed 14 banding patterns. Based on the 16S rDNA gene sequence, the isolates were identified as Lactobacillus plantarum (75.7%), Weissella cibaria (13.5%), Lactobacillus brevis (8.1%), and Weissella confusa (2.7%). Phenotypic characterization showed that most of the isolates were homofermentative bacilli, able to ferment glucose, maltose, cellobiose, and fructose and grow in a broad range of temperatures and pH. The isolates were highly susceptible to ampicillin, amoxicillin, clindamycin, chloramphenicol, erythromicyn, penicillin, and tetracycline and showed great resistance to kanamycin, gentamycin, streptomycin, sulfamethoxazole/trimethoprim, and vancomycin. No proteolytic or amylolytic activity was detected. L. plantarum strains produce lactic acid in higher concentrations and Weissella strains produce exopolymers only from sucrose. Molecular methods allowed to accurately identify the LAB isolates from the Peruvian Amazonian fruits, while phenotypic methods provided information about their metabolism, physiology and other characteristics that may be useful in future biotechnological processes. Further research will focus especially on the study of L. plantarum strains.The objective of this research was the identification and characterization of lactic acid bacteria (LAB) isolated from Peruvian Amazonian fruits. Thirty-seven isolates were obtained from diverse Amazonian fruits. Molecular characterization of the isolates was performed by ARDRA, 16S-23S ITS RFLP and rep-PCR using GTG5 primers. Identification was carried out by sequencing the 16S rDNA gene. Phenotypic characterization included nutritional, physiological and antimicrobial resistance tests. Molecular characterization by Amplified Ribosomal DNA Restriction Analysis (ARDRA) and 16S-23S ITS RFLP resulted in four restriction profiles while GTG5 analysis showed 14 banding patterns. Based on the 16S rDNA gene sequence, the isolates were identified as Lactobacillus plantarum (75.7%), Weissella cibaria (13.5%), Lactobacillus brevis (8.1%), and Weissella confusa (2.7%). Phenotypic characterization showed that most of the isolates were homofermentative bacilli, able to ferment glucose, maltose, cellobiose, and fructose and grow in a broad range of temperatures and pH. The isolates were highly susceptible to ampicillin, amoxicillin, clindamycin, chloramphenicol, erythromicyn, penicillin, and tetracycline and showed great resistance to kanamycin, gentamycin, streptomycin, sulfamethoxazole/trimethoprim, and vancomycin. No proteolytic or amylolytic activity was detected. L. plantarum strains produce lactic acid in higher concentrations and Weissella strains produce exopolymers only from sucrose. Molecular methods allowed to accurately identify the LAB isolates from the Peruvian Amazonian fruits, while phenotypic methods provided information about their metabolism, physiology and other characteristics that may be useful in future biotechnological processes. Further research will focus especially on the study of L. plantarum strains.
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