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Redox Biology Jun 2024Salmonella infection entails a cascade of attacks and defence measures. After breaching the intestinal epithelial barrier, Salmonella is phagocytosed by macrophages,...
Salmonella Typhimurium employs spermidine to exert protection against ROS-mediated cytotoxicity and rewires host polyamine metabolism to ameliorate its survival in macrophages.
Salmonella infection entails a cascade of attacks and defence measures. After breaching the intestinal epithelial barrier, Salmonella is phagocytosed by macrophages, where the bacteria encounter multiple stresses, to which it employs relevant countermeasures. Our study shows that, in Salmonella, the polyamine spermidine activates a stress response mechanism by regulating critical antioxidant genes. Salmonella Typhimurium mutants for spermidine transport and synthesis cannot mount an antioxidative response, resulting in high intracellular ROS levels. These mutants are also compromised in their ability to be phagocytosed by macrophages. Furthermore, it regulates a novel enzyme in Salmonella, Glutathionyl-spermidine synthetase (GspSA), which prevents the oxidation of proteins in E. coli. Moreover, the spermidine mutants and the GspSA mutant show significantly reduced survival in the presence of hydrogen peroxide in vitro and reduced organ burden in the mouse model of Salmonella infection. Conversely, in macrophages isolated from gp91phox mice, we observed a rescue in the attenuated fold proliferation previously observed upon infection. We found that Salmonella upregulates polyamine biosynthesis in the host through its effectors from SPI-1 and SPI-2, which addresses the attenuated proliferation observed in spermidine transport mutants. Thus, inhibition of this pathway in the host abrogates the proliferation of Salmonella Typhimurium in macrophages. From a therapeutic perspective, inhibiting host polyamine biosynthesis using an FDA-approved chemopreventive drug, D, L-α-difluoromethylornithine (DFMO), reduces Salmonella colonisation and tissue damage in the mouse model of infection while enhancing the survival of infected mice. Therefore, our work provides a mechanistic insight into the critical role of spermidine in stress resistance of Salmonella. It also reveals a bacterial strategy in modulating host metabolism to promote their intracellular survival and shows the potential of DFMO to curb Salmonella infection.
Topics: Animals; Salmonella typhimurium; Spermidine; Mice; Macrophages; Reactive Oxygen Species; Bacterial Proteins; Polyamines; Phagocytosis; Salmonella Infections; NADPH Oxidases; Host-Pathogen Interactions; Spermidine Synthase; Oxidative Stress; NADPH Oxidase 2; Membrane Proteins
PubMed: 38593631
DOI: 10.1016/j.redox.2024.103151 -
Journal of Medicinal Chemistry Apr 2024Cisplatin (cDDP) resistance is a matter of concern in triple-negative breast cancer therapeutics. We measured the metabolic response of cDDP-sensitive (S) and -resistant...
Cisplatin (cDDP) resistance is a matter of concern in triple-negative breast cancer therapeutics. We measured the metabolic response of cDDP-sensitive (S) and -resistant (R) MDA-MB-231 cells to PdSpermine(Spm) (a possible alternative to cDDP) compared to cDDP to investigate (i) intrinsic response/resistance mechanisms and (ii) the potential cytotoxic role of PdSpm. Cell extracts were analyzed by untargeted nuclear magnetic resonance metabolomics, and cell media were analyzed for particular metabolites. CDDP-exposed S cells experienced enhanced antioxidant protection and small deviations in the tricarboxylic acid cycle (TCA), pyrimidine metabolism, and lipid oxidation (proposed cytotoxicity signature). R cells responded more strongly to cDDP, suggesting a resistance signature of activated TCA cycle, altered AMP/ADP/ATP and adenine/uracil fingerprints, and phospholipid biosynthesis (without significant antioxidant protection). PdSpm impacted more markedly on R/S cell metabolisms, inducing similarities to cDDP/S cells (probably reflecting high cytotoxicity) and strong additional effects indicative of amino acid depletion, membrane degradation, energy/nucleotide adaptations, and a possible beneficial intracellular γ-aminobutyrate/glutathione-mediated antioxidant mechanism.
Topics: Humans; Triple Negative Breast Neoplasms; Cisplatin; Drug Resistance, Neoplasm; Antineoplastic Agents; Cell Line, Tumor; Female; Spermine; Palladium
PubMed: 38590144
DOI: 10.1021/acs.jmedchem.4c00435 -
The Journal of Biological Chemistry May 2024Spermine synthase is an aminopropyltransferase that adds an aminopropyl group to the essential polyamine spermidine to form tetraamine spermine, needed for normal human...
Spermine synthase is an aminopropyltransferase that adds an aminopropyl group to the essential polyamine spermidine to form tetraamine spermine, needed for normal human neural development, plant salt and drought resistance, and yeast CoA biosynthesis. We functionally identify for the first time bacterial spermine synthases, derived from phyla Bacillota, Rhodothermota, Thermodesulfobacteriota, Nitrospirota, Deinococcota, and Pseudomonadota. We also identify bacterial aminopropyltransferases that synthesize the spermine same mass isomer thermospermine, from phyla Cyanobacteriota, Thermodesulfobacteriota, Nitrospirota, Dictyoglomota, Armatimonadota, and Pseudomonadota, including the human opportunistic pathogen Pseudomonas aeruginosa. Most of these bacterial synthases were capable of synthesizing spermine or thermospermine from the diamine putrescine and so possess also spermidine synthase activity. We found that most thermospermine synthases could synthesize tetraamine norspermine from triamine norspermidine, that is, they are potential norspermine synthases. This finding could explain the enigmatic source of norspermine in bacteria. Some of the thermospermine synthases could synthesize norspermidine from diamine 1,3-diaminopropane, demonstrating that they are potential norspermidine synthases. Of 18 bacterial spermidine synthases identified, 17 were able to aminopropylate agmatine to form N-aminopropylagmatine, including the spermidine synthase of Bacillus subtilis, a species known to be devoid of putrescine. This suggests that the N-aminopropylagmatine pathway for spermidine biosynthesis, which bypasses putrescine, may be far more widespread than realized and may be the default pathway for spermidine biosynthesis in species encoding L-arginine decarboxylase for agmatine production. Some thermospermine synthases were able to aminopropylate N-aminopropylagmatine to form N-guanidinothermospermine. Our study reveals an unsuspected diversification of bacterial polyamine biosynthesis and suggests a more prominent role for agmatine.
Topics: Bacteria; Bacterial Proteins; Spermidine; Spermidine Synthase; Spermine; Spermine Synthase; Polyamines; Alkyl and Aryl Transferases; Agmatine
PubMed: 38588807
DOI: 10.1016/j.jbc.2024.107281 -
Advanced Science (Weinheim,... Jun 2024Acid-induced arginine decarboxylase AdiA is a typical homo-oligomeric protein biosynthesizing alkaline nylon monomer putrescine. However, upon loss of the AdiA decamer...
Acid-induced arginine decarboxylase AdiA is a typical homo-oligomeric protein biosynthesizing alkaline nylon monomer putrescine. However, upon loss of the AdiA decamer oligomeric state at neutral and alkaline conditions the activity also diminishes, obstructing the whole-cell biosynthesis of alkaline putrescine. Here, a structure cohesion strategy is proposed to change the pH adaptation of AdiA to alkaline environments based on the rational engineering of meridional and latitudinal oligomerization interfaces. After integrating substitutions of E467K at the latitudinal interface and H736E at the meridional channel interface, the structural stability of AdiA decamer and its substrate transport efficiency at neutral and alkaline conditions are improved. Finally, E467K_H736E is well adapted to neutral and alkaline environments (pH 7.0-9.0), and its enzymatic activity is 35-fold higher than that of wild AdiA at pH 8.0. Using E467K_H736E in the putrescine synthesis pathway, the titer of putrescine is up to 128.9 g·L with a conversion of 0.94 mol·mol in whole-cell catalysis. Additionally, the neutral pH adaptation of lysine decarboxylase, with a decamer structure similar to AdiA, is also improved using this cohesion strategy, providing an option for pH-adaptation engineering of other oligomeric decarboxylases.
Topics: Carboxy-Lyases; Hydrogen-Ion Concentration; Escherichia coli; Putrescine
PubMed: 38569221
DOI: 10.1002/advs.202307779 -
MSystems May 2024Dietary fiber deprivation is linked to probiotic extinction, mucus barrier dysbiosis, and the overgrowth of mucin-degrading bacteria. However, whether and how mucin...
UNLABELLED
Dietary fiber deprivation is linked to probiotic extinction, mucus barrier dysbiosis, and the overgrowth of mucin-degrading bacteria. However, whether and how mucin could rescue fiber deprivation-induced intestinal barrier defects remains largely unexplored. Here, we sought to investigate the potential role and mechanism by which exogenous mucin maintains the gut barrier function. The results showed that dietary mucin alleviated fiber deprivation-induced disruption of colonic barrier integrity and reduced spermine production . Importantly, we highlighted that microbial-derived spermine production, but not host-produced spermine, increased significantly after mucin supplementation, with a positive association with upgraded colonic abundance. After employing an model, the microbial-derived spermine was consistently dominated by both mucin and spp. Furthermore, was identified as an essential spermine-producing spp., and this isolated strain was responsible for spermine accumulation, especially after adhering to mucin . Specifically, the mucin-supplemented bacterial supernatant of was verified to promote intestinal barrier functions through the increased spermine production with a dependence on enhanced arginine metabolism. Overall, these findings collectively provide evidence that mucin-modulated microbial arginine metabolism bridged the interplay between microbes and gut barrier function, illustrating possible implications for host gut health.
IMPORTANCE
Microbial metabolites like short-chain fatty acids produced by dietary fiber fermentation have been demonstrated to have beneficial effects on intestinal health. However, it is essential to acknowledge that certain amino acids entering the colon can be metabolized by microorganisms to produce polyamines. The polyamines can promote the renewal of intestinal epithelial cell and maintain host-microbe homeostasis. Our study highlighted the specific enrichment by mucin on promoting the arginine metabolism in to produce spermine, suggesting that microbial-derived polyamines support a significant enhancement on the goblet cell proliferation and barrier function.
Topics: Spermine; Mucins; Arginine; Intestinal Mucosa; Animals; Gastrointestinal Microbiome; Colon; Male; Mice; Lactobacillus; Humans; Dietary Fiber; Mice, Inbred C57BL
PubMed: 38564708
DOI: 10.1128/msystems.00246-24 -
MicrobiologyOpen Apr 2024Arginine-ornithine metabolism plays a crucial role in bacterial homeostasis, as evidenced by numerous studies. However, the utilization of arginine and the downstream...
Arginine-ornithine metabolism plays a crucial role in bacterial homeostasis, as evidenced by numerous studies. However, the utilization of arginine and the downstream products of its metabolism remain undefined in various gut bacteria. To bridge this knowledge gap, we employed genomic screening to pinpoint relevant metabolic targets. We also devised a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics method to measure the levels of arginine, its upstream precursors, and downstream products in cell-free conditioned media from enteric pathobionts, including Escherichia coli, Klebsiella aerogenes, K. pneumoniae, Pseudomonas fluorescens, Acinetobacter baumannii, Streptococcus agalactiae, Staphylococcus epidermidis, S. aureus, and Enterococcus faecalis. Our findings revealed that all selected bacterial strains consumed glutamine, glutamate, and arginine, and produced citrulline, ornithine, and GABA in our chemically defined medium. Additionally, E. coli, K. pneumoniae, K. aerogenes, and P. fluorescens were found to convert arginine to agmatine and produce putrescine. Interestingly, arginine supplementation promoted biofilm formation in K. pneumoniae, while ornithine supplementation enhanced biofilm formation in S. epidermidis. These findings offer a comprehensive insight into arginine-ornithine metabolism in enteric pathobionts.
Topics: Ornithine; Putrescine; Arginine; Escherichia coli; Chromatography, Liquid; Staphylococcus aureus; Tandem Mass Spectrometry; Bacteria; Klebsiella pneumoniae
PubMed: 38560776
DOI: 10.1002/mbo3.1408 -
Clinical and Translational Medicine Apr 2024
Topics: Humans; Astrocytes; gamma-Aminobutyric Acid; Brain Diseases
PubMed: 38558537
DOI: 10.1002/ctm2.1642 -
Scientific Reports Mar 2024An extreme thermophilic bacterium, Thermus thermophilus produces more than 20 unusual polyamines, but their biosynthetic pathways, including homospermidine, are not yet...
An extreme thermophilic bacterium, Thermus thermophilus produces more than 20 unusual polyamines, but their biosynthetic pathways, including homospermidine, are not yet fully understood. Two types of homospermidine synthases have been identified in plants and bacteria, which use spermidine and putrescine or two molecules of putrescine as substrates. However, homospermidine synthases with such substrate specificity have not been identified in T. thermophilus. Here we identified a novel agmatine homocoupling enzyme that is involved in homospermidine biosynthesis in T. thermophilus. The reaction mechanism is different from that of a previously described homospermidine synthase, and involves conjugation of two molecules of agmatine, which produces a diamidino derivative of homospermidine (caldomycin) as an immediate precursor of homospermidine. We conclude that there is a homospermidine biosynthetic pathway from agmatine via caldomycin synthase followed by ureohydrolase in T. thermophilus. Furthermore, it is shown that caldomycin is a novel compound existing in nature.
Topics: Putrescine; Agmatine; Polyamines; Spermidine; Plants
PubMed: 38555406
DOI: 10.1038/s41598-024-58296-0 -
Foods (Basel, Switzerland) Mar 2024Dill ( L.) essential oil (DEO) obtained by hydrodistillation (HD) and lipid extracts (DSE and DSE) obtained by supercritical CO extraction (SFE) were used as potential...
Dill ( L.) essential oil (DEO) obtained by hydrodistillation (HD) and lipid extracts (DSE and DSE) obtained by supercritical CO extraction (SFE) were used as potential antioxidants and antimicrobial agents in beef burgers at two different concentrations (0.075 and 0.15 µL/g). The chemical profile of the lipid extracts and their in vitro antimicrobial activity against the common pathogens and (MIC and MBC) were determined. The quality and shelf life of the burgers were monitored through (lipid oxidation-TBARS test; protein oxidation-thiol group content and selected biogenic amine content) and microbiological quality (-EB, aerobic mesophilic bacteria-TAMB, lactic acid bacteria-LAB). Dill lipid extracts (DEO and DSE) significantly ( < 0.05) reduced lipid oxidation and protein oxidation in beef burgers, while the lipid extract (DSE) showed pro-oxidative effects. The strongest antimicrobial potential against EB was found in SFE150 (1.15 log cfu/g). Putrescine, cadaverine, histamine, and tyramine were not detected in any of the analyzed samples during the storage period, while the total content of biogenic amines ranged from 21.4 mg/kg to 285 mg/kg. Generally, it can be concluded that dill essential oil (DEO) and extract DSE can be used as novel natural additives in minced-meat products.
PubMed: 38540886
DOI: 10.3390/foods13060896 -
Foods (Basel, Switzerland) Mar 2024The influence of different levels of sodium chloride, sodium nitrite, and glucose on the quality characteristics of spontaneously fermented goat meat sausage was...
The influence of different levels of sodium chloride, sodium nitrite, and glucose on the quality characteristics of spontaneously fermented goat meat sausage was investigated. The amounts of total biogenic amines in all the sausages ranged from 324.70 to 388.77 mg/kg; among them, spermine was the most abundant, with amounts ranging from 230.96 to 275.78 mg/kg. Increasing sodium chloride from 15 to 35 g/kg, the content of cadaverine, putrescine, tyramine, phenylethylamine, tryptamine, and total amines decreased, and Enterobacteriaceae counts decreased at the same time. Increasing sodium nitrite from 150 to 250 mg/kg, the content of cadaverine, histamine, and total amines decreased, while Enterobacteriaceae counts decreased simultaneously. Increasing glucose from 10 to 40 g/kg, the content of cadaverine, spermidine, and total amines decreased. was the most abundant genus across all the samples, and the relative abundance of was reduced obviously by increasing sodium nitrite and glucose levels. The top 10 differential bacterial taxa for each additive group were respectively obtained, and microbial biomarkers for each level of additive within its group were acquired, respectively. Through Pearson correlation, was positively correlated with phenylethylamine, tryptamine, tyramine, and cadaverine, and were positively correlated with phenylethylamine and putrescine, respectively, suggesting they have the potential to produce biogenic amines. The results provided references for controlling the accumulation of biogenic amines in fermented goat meat sausage via the addition of auxiliary additives during the processing.
PubMed: 38540808
DOI: 10.3390/foods13060817