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FEBS Letters Jun 1992The capacity of microsomes from aminopyrine-induced Jerusalem artichoke (Helianthus tuberosus L.) to oxidize saturated and unsaturated fatty acids has been investigated...
The capacity of microsomes from aminopyrine-induced Jerusalem artichoke (Helianthus tuberosus L.) to oxidize saturated and unsaturated fatty acids has been investigated using lauric acid and a series of unsaturated lauric acid analogs (7-, 8-, 9- and 10-dodecenoic acids) as radiolabeled substrates. In the presence of NADH, lauric acid was mono-hydroxylated principally at carbon 9. Steric analysis of this product showed a low enantiomeric excess of 28%. Mono-hydroxylated and mono-epoxidated reaction products were formed from the unsaturated analogs. The epoxidation/hydroxylation ratio was related to the position of the double bond in the aliphatic chain. The oxidation of 7-dodecenoic acid (7-DDNA) and 10-DDNA produced mainly 9-hydroxy-7-DDNA and 9-hydroxy-10-DDNA plus minor amounts of 7,8-epoxy- or 10,11-epoxylauric acid, respectively. In contrast, 8- and 9-DDNAs yielded essentially 8,9-epoxy- and 9,10-epoxylauric acids and smaller amounts of 10-hydroxy-9-DDNA and 8-hydroxy-9-DDNA, respectively. The optical purity and the absolute configuration of the major metabolites were investigated. Epoxidation of Z 8-DDNA and Z 9-DDNA occurs with high enantiomeric excesses. When the double bond was in the Z configuration, (8S,9R)/(8R,9S) 8,9-epoxylauric acid (93/7) or (9R,10S)/(9S,10R) 9,10-epoxylauric acid (89/11) were produced. In contrast, when the double bond was in the E configuration, steric analysis showed an enantiomeric ratio of 52/48 for E 8,9-epoxide and of 59/41 for E 9,10-epoxide. Z 7-DDNA led to the formation of 98% of the 9(S)-hydroxy-Z 7-DDNA enantiomer, while 9-hydroxy-Z 10-DDNA derived from Z 10-DDNA was 35% (R) and 65% (S).
Topics: Fatty Acids; Lauric Acids; Microsomes; Oxidation-Reduction; Oxygen; Plants; Stereoisomerism
PubMed: 1607005
DOI: 10.1016/0014-5793(92)80499-7 -
BMC Microbiology Feb 2014Extensive use of antibiotics has fostered the emergence of superbugs that are resistant to multidrugs, which becomes a great healthcare and public concern. Previous...
Diffusible signal factor (DSF) quorum sensing signal and structurally related molecules enhance the antimicrobial efficacy of antibiotics against some bacterial pathogens.
BACKGROUND
Extensive use of antibiotics has fostered the emergence of superbugs that are resistant to multidrugs, which becomes a great healthcare and public concern. Previous studies showed that quorum sensing signal DSF (diffusible signal factor) not only modulates bacterial antibiotic resistance through intraspecies signaling, but also affects bacterial antibiotic tolerance through interspecies communication. These findings motivate us to exploit the possibility of using DSF and its structurally related molecules as adjuvants to influence antibiotic susceptibility of bacterial pathogens.
RESULTS
In this study, we have demonstrated that DSF signal and its structurally related molecules could be used to induce bacterial antibiotic susceptibility. Exogenous addition of DSF signal (cis-11-methyl-2-dodecenoic acid) and its structural analogues could significantly increase the antibiotic susceptibility of Bacillus cereus, possibly through reducing drug-resistant activity, biofilm formation and bacterial fitness. The synergistic effect of DSF and its structurally related molecules with antibiotics on B. cereus is dosage-dependent. Combination of DSF with gentamicin showed an obviously synergistic effect on B. cereus pathogenicity in an in vitro model. We also found that DSF could increase the antibiotic susceptibility of other bacterial species, including Bacillus thuringiensis, Staphylococcus aureus, Mycobacterium smegmatis, Neisseria subflava and Pseudomonas aeruginosa.
CONCLUSION
The results indicate a promising potential of using DSF and its structurally related molecules as novel adjuvants to conventional antibiotics for treatment of infectious diseases caused by bacterial pathogens.
Topics: Anti-Bacterial Agents; Dose-Response Relationship, Drug; Drug Synergism; Gentamicins; Gram-Negative Bacteria; Gram-Positive Bacteria; Humans; Lauric Acids; Quorum Sensing
PubMed: 24575808
DOI: 10.1186/1471-2180-14-51 -
The ISME Journal Jan 2008In addition to producing lethal antibiotics, microorganisms may also use a new form of antagonistic mechanism in which signal molecules are exported to influence the...
In addition to producing lethal antibiotics, microorganisms may also use a new form of antagonistic mechanism in which signal molecules are exported to influence the gene expression and hence the ecological competence of their competitors. We report here the isolation and characterization of a novel signaling molecule, cis-2-dodecenoic acid (BDSF), from Burkholderia cenocepacia. BDSF is structurally similar to the diffusible signal factor (DSF) that is produced by the RpfF enzyme of Xanthomonas campestris. Deletion analysis demonstrated that Bcam0581, which encodes an RpfF homologue, was essential for BDSF production. The gene is highly conserved and widespread in the Burkholderia cepacia complex. Exogenous addition of BDSF restored the biofilm and extracellular polysaccharide production phenotypes of Xanthomonas campestris pv. campestris DSF-deficient mutants, highlighting its potential role in inter-species signaling. Further analyses showed that Candida albicans germ tube formation was strongly inhibited by either coculture with B. cenocepacia or by exogenous addition of physiological relevant levels of BDSF, whereas deletion of Bcam0581 abrogated the inhibitory ability of the bacterial pathogen. As B. cenocepacia and C. albicans are frequently encountered human pathogens, identification of the BDSF signal and its activity thus provides a new insight into the molecular grounds of their antagonistic interactions whose importance to microbial ecology and pathogenesis is now becoming evident.
Topics: Antibiosis; Bacterial Proteins; Biofilms; Burkholderia Infections; Burkholderia cepacia; Candida albicans; Enoyl-CoA Hydratase; Fatty Acids, Monounsaturated; Gene Expression Regulation, Bacterial; Genetic Complementation Test; Mutant Proteins; Open Reading Frames; Polysaccharides, Bacterial; Sequence Alignment; Sequence Deletion; Signal Transduction; Xanthomonas campestris
PubMed: 18049456
DOI: 10.1038/ismej.2007.76 -
The Journal of Biological Chemistry Aug 1995The intermediate metabolic events which degrade hydroxy polyunsaturated fatty acids is largely unknown. Such molecules are common products of lipid peroxidation and...
Metabolism of 6-trans-isomers of leukotriene B4 in cultured hepatoma cells and in human polymorphonuclear leukocytes. Identification of a delta 6-reductase metabolic pathway.
The intermediate metabolic events which degrade hydroxy polyunsaturated fatty acids is largely unknown. Such molecules are common products of lipid peroxidation and lipoxygenase catalyzed oxidation of arachidonic acid. Metabolism of two 5,12-dihydroxyeicosatetraenoic acids, 6-trans-LTB4 (leukotriene B4), and 6-trans-12-epi-LTB4 was studied in HepG2 cells (a human-derived hepatoma cell line). Extensive metabolism was observed with a major metabolite identified as 4-hydroxy-6-dodecenoic acid for both epimers. Incubation of 6-trans-LTB4 epimers at shorter times revealed the formation of intermediate metabolites, including 6-hydroxy-4,8-tetradecadienoic acid and 8-hydroxy-4,6,10-hexadecatrienoic acid suggesting beta-oxidation as the major pathway leading to the formation of the common terminal metabolite. Two additional metabolites were structurally elucidated as 5-oxo-6,7-dihydro-LTB4 and 6,7-dihydro-LTB4 which have not been previously described. Formation of 5-oxo-6,7-dihydro-LTB4 and 6,7-dihydro-LTB4 were also observed during metabolism of 6-trans-12-epi-LTB4 in human polymorphonuclear leukocytes. Of particular interest is the metabolism of these compounds by beta-oxidation from the carboxyl terminus, a process which is not observed with leukotriene B4 or leukotriene C4. Identification of these metabolites suggested the operation of the 5-hydroxyeicosanoid dehydrogenase pathway followed by a delta 6-reductase metabolic pathway which has not been previously described. This pathway of beta-oxidation may limit the activity of various 5,12-diHETEs including nonenzymatic hydrolysis products of LTA4 and also the recently described B4-isoleukotrienes.
Topics: Carcinoma, Hepatocellular; Humans; In Vitro Techniques; Leukotriene B4; Mass Spectrometry; Neutrophils; Oxidation-Reduction; Oxidoreductases; Stereoisomerism; Tumor Cells, Cultured
PubMed: 7649996
DOI: 10.1074/jbc.270.34.19845 -
Microbiology (Reading, England) Mar 2008Exopolysaccharide and several extracellular enzymes of Xanthomonas campestris pv. campestris (Xcc), the causative agent of black rot in crucifers, are virulence factors....
Exopolysaccharide and several extracellular enzymes of Xanthomonas campestris pv. campestris (Xcc), the causative agent of black rot in crucifers, are virulence factors. In this study, sequence and mutational analysis has demonstrated that Xcc pehA encodes the major polygalacturonase, a member of family 28 of the glycosyl hydrolases. Using the 5' RACE (rapid amplification of cDNA ends) method, the pehA transcription initiation site was mapped at 102 nt downstream of a Clp (cAMP receptor protein-like protein)-binding site. Transcriptional fusion assays showed that pehA transcription is greatly induced by polygalacturonic acid, positively regulated by Clp and RpfF (an enoyl-CoA hydratase homologue which is required for the synthesis of cis-11-methyl-2-dodecenoic acid, a low-molecular-mass diffusible signal factor), subjected to catabolite repression, which is independent of Clp or RpfF, and repressed under conditions of oxygen limitation or nitrogen starvation. Our findings extend previous work on Clp and RpfF regulation to show that they both influence the expression of pehA in Xcc.
Topics: Artificial Gene Fusion; Bacterial Proteins; Base Sequence; Endopeptidase Clp; Enoyl-CoA Hydratase; Gene Expression Regulation, Bacterial; Genes, Reporter; Molecular Sequence Data; Pectins; Polygalacturonase; Promoter Regions, Genetic; Transcription Initiation Site; Xanthomonas campestris; beta-Galactosidase
PubMed: 18310017
DOI: 10.1099/mic.0.2007/012930-0 -
Chemical Research in Toxicology May 2007Polyunsaturated fatty acids can be converted to lipid hydroperoxides through nonenzymatic and enzymatic pathways. The prototypic omega-6 lipid hydroperoxide...
Polyunsaturated fatty acids can be converted to lipid hydroperoxides through nonenzymatic and enzymatic pathways. The prototypic omega-6 lipid hydroperoxide 13-hydroperoxy-octadecadienoic acid (13-HPODE) homolytically decomposes to form highly reactive alpha,beta-unsaturated aldehydes, such as 9,12-dioxo-10(E)-dodecenoic acid (DODE), 4-oxo-2(E)-nonenal (ONE), 4,5-epoxy-2(E)-decenal (EDE), and 4-hydroxy-2(E)-nonenal (HNE), that can form covalent adducts with DNA. Both 4-oxo-2(E)-nonenal and 4-hydroxy-2(E)-nonenal can also modify proteins to form products that can potentially serve as biomarkers of lipid hydroperoxide-mediated macromolecule damage. In this study, cytochrome c was used to identify and individually characterize the modification sites for each of these aldehydes and also determine the most abundant adduct formed following the decomposition of 13-HPODE. The adducts were characterized by ESI-TOF/MS analysis of the intact proteins and by a combination of ESI-ion-trap/MSn and quadrupole-TOF/MS/MS analysis of the tryptic and chymotryptic peptides. The major adducts included an HNE-His Michael adduct on H33, EDE-Lys adducts on K7 and K8, ONE-Lys ketoamide adducts on K5, K7, and K8, an apparent ONE-Lys Michael adduct on K5, and DODE-Lys carboxyl ketoamide adducts on K86 and K87. DODE was the most reactive aldehyde toward cytochrome c. The major adduct from this reaction was analogous to the most abundant adduct resulting from the decomposition of 13-HPODE in the presence of cytochrome c.
Topics: Cytochromes c; DNA Adducts; DNA Damage; Linoleic Acids; Lipid Peroxides; Myoglobin; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry
PubMed: 17407328
DOI: 10.1021/tx600289r -
Research in Microbiology 2007Stenotrophomonas maltophilia is widespread in natural environments such as soil, sewage and plant rhizospheres. Surfactants frequently function in modulating bacterial...
Stenotrophomonas maltophilia is widespread in natural environments such as soil, sewage and plant rhizospheres. Surfactants frequently function in modulating bacterial surface translocation. In this study, rpfB and rpfF orthologues were identified from S. maltophilia strain WR-C, which was isolated from the clogged zone of a septic system. These genes play a role in the biosynthesis of eight extracellular compounds that facilitated flagella-independent translocation by the wild-type or a flagella-defective mutant. This type of surface translocation has not been reported previously for this organism. These eight compounds include cis-delta 2-11-methyl-dodecenoic acid and seven structural derivatives. Two are saturated fatty acids; the others are unsaturated fatty acids with double bonds at position 2. These fatty acids vary in chain length from 12 to 14 carbons and in the position of the branched methyl group. Our results demonstrated that independently cis-delta 2-11-methyl-dodecenoic acid and 11-methyl-dodecanoic acid promoted flagella-independent translocation by S. maltophilia strain WR-C by acting as wetting agents.
Topics: Aconitate Hydratase; Bacterial Proteins; Biological Transport; Cytokines; Fatty Acids; Flagella; Stenotrophomonas maltophilia
PubMed: 18054205
DOI: 10.1016/j.resmic.2007.09.002 -
Scientific Reports Aug 2015Members of the diffusible signal factor (DSF) family are a novel class of quorum sensing (QS) signals in diverse Gram-negative bacteria. Although previous studies have...
Members of the diffusible signal factor (DSF) family are a novel class of quorum sensing (QS) signals in diverse Gram-negative bacteria. Although previous studies have identified RpfF as a key enzyme for the biosynthesis of DSF family signals, many questions in their biosynthesis remain to be addressed. In this study with the phytopathogen Xanthomonas campestris pv. campestris (Xcc), we show that Xcc produces four DSF-family signals (DSF, BDSF, CDSF and IDSF) during cell culture, and that IDSF is a new functional signal characterized as cis-10-methyl-2-dodecenoic acid. Using a range of defined media, we further demonstrate that Xcc mainly produces BDSF in the presence of carbohydrates; leucine and valine are the primary precursor for DSF biosynthesis; isoleucine is the primary precursor for IDSF biosynthesis. Furthermore, our biochemical analyses show that the key DSF synthase RpfF has both thioesterase and dehydratase activities, and uses 3-hydroxydedecanoyl-ACP as a substrate to produce BDSF. Finally, our results show that the classic fatty acid synthesis elongation cycle is required for the biosynthesis of DSF-family signals. Taken all together, these findings establish a general biosynthetic pathway for the DSF-family quorum sensing signals.
Topics: Amino Acids, Branched-Chain; Bacterial Proteins; Biosynthetic Pathways; Carbohydrates; Cerulenin; Diffusion; Fatty Acids; Genes, Bacterial; Models, Biological; Quorum Sensing; Thiolester Hydrolases; Xanthomonas campestris
PubMed: 26289160
DOI: 10.1038/srep13294 -
Journal of Chemical Ecology Jan 2019Two heliozelid species, Antispila oinophylla van Nieukerken & Wagner and Holocacista rivillei (Stainton) severely infest Italian grapevines. The volatile pheromones from...
Two heliozelid species, Antispila oinophylla van Nieukerken & Wagner and Holocacista rivillei (Stainton) severely infest Italian grapevines. The volatile pheromones from calling females were collected by solid phase micro extraction (SPME) and analyzed by gas chromatography with electroantennographic detection (GC-EAD). Two compounds from A. oinophylla females eliciting electrophysiological activity from the conspecific male antenna were identified as (Z)-5-tetradecenal and (Z)-7-tetradecenal by coupled gas chromatography/mass spectrometry (GC/MS) analysis. SPME collections from H. rivillei produced no GC-EAD active compounds but analysis of fatty acyl moieties in the pheromone gland, demonstrated the presence of the putative pheromone biosynthetic precursors (Z)-5-dodecenoic acid and (Z)-7-tetradecenoic acid. Field trapping experiments in Italy confirmed that (Z)-5-tetradecenal and (Z)-7-tetradecenal are essential for the attraction of male A. oinophylla in a blend ratio of 15:100 respectively, whereas (Z)-5-dodecenal and (Z)-7-tetradecenal attract male H. rivillei in a blend ratio of 100:6.
Topics: Aldehydes; Animals; Female; Gas Chromatography-Mass Spectrometry; Lepidoptera; Male; Sex Attractants; Vitis
PubMed: 30547362
DOI: 10.1007/s10886-018-1036-z -
Journal of Bacteriology Dec 2009Burkholderia cenocepacia produces the molecule cis-2-dodecenoic acid (BDSF), which was previously shown to play a role in antagonism against the fungal pathogen Candida...
Burkholderia cenocepacia produces the molecule cis-2-dodecenoic acid (BDSF), which was previously shown to play a role in antagonism against the fungal pathogen Candida albicans by interfering with its morphological transition. In this study, we show that production of BDSF is under stringent transcriptional control and the molecule accumulates in a cell density-dependent manner, typically found with quorum-sensing (QS) signals. B. cenocepacia mutant strain J2315 with a deleted Bcam0581 gene, which encodes an enzyme essential for BDSF production, exhibited a growth defect in minimal medium but not in rich medium, decreased virulence gene expression, and attenuated virulence in a zebrafish infection model. Exogenous addition of BDSF to the mutant rescues virulence gene expression but fails to restore its growth defect in minimal medium. We show that Bcam0581, but not BDSF, is associated with B. cenocepacia ATP biogenesis. We also provide evidence that some of the BDSF-regulated genes are also controlled by the acyl-homoserine-lactone-dependent QS system and are thus coregulated by two cell-to-cell signaling systems. These data demonstrate that in addition to the role in cross-kingdom signal interference, BDSF and its synthase are also important for the virulence and physiology of B. cenocepacia.
Topics: Adenosine Triphosphate; Animals; Bacterial Proteins; Burkholderia; Energy Metabolism; Fatty Acids, Monounsaturated; Gene Expression Regulation, Bacterial; Quorum Sensing; Reverse Transcriptase Polymerase Chain Reaction; Virulence; Zebrafish
PubMed: 19801414
DOI: 10.1128/JB.00681-09