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Veillonella spp. are nitrate-reducing bacteria with anaerobic respiratory activity that reduce nitrate to nitrite. They are obligate anaerobic, Gram-negative cocci that ferment lactate as the main carbon source and produce short-chain fatty acids (SCFAs). Commensal Veillonella reside in the human body site where lactate level is, however, limited for Veillonella growth. In this study, nitrate was shown to promote the anaerobic growth of Veillonella in the lactate-deficient media. We aimed to investigate the underlying mechanisms and the metabolism involved in nitrate respiration. Nitrate (15 mM) was demonstrated to promote Veillonella dispar growth and viability in the tryptone-yeast extract medium containing 0.5 mM L-lactate. Metabolite and transcriptomic analyses revealed nitrate enabled V. dispar to actively utilize glutamate and aspartate from the medium and secrete tryptophan. Glutamate or aspartate was further supplemented to a medium to investigate individual catabolism during nitrate respiration. Notably, nitrate was demonstrated to elevate SCFA production in the glutamate-supplemented medium, and further increase tryptophan production in the aspartate-supplemented medium. We proposed that the increased consumption of glutamate provided reducing power for nitrate respiration and aspartate served as a substrate for fumarate formation. Both glutamate and aspartate were incorporated into the central metabolic pathways via reverse tricarboxylic acid cycle and were linked with the increased production of acetate, propionate, and tryptophan. This study provides further understanding of the promoted growth and metabolic mechanisms by commensal V. dispar utilizing nitrate and specific amino acids to adapt to the lactate-deficient environment.IMPORTANCENitrate is a pivotal ecological factor influencing microbial community and metabolism. Dietary nitrate provides health benefits including anti-diabetic and anti-hypertensive effects via microbial-derived metabolites such as nitrite. Unraveling the impacts of nitrate on the growth and metabolism of human commensal bacteria is imperative to comprehend the intricate roles of nitrate in regulating microbial metabolism, community, and human health. Veillonella are lactate-utilizing, nitrate-reducing bacteria that are frequently found in the human body site where lactate levels are low and nitrate is at millimolar levels. Here, we comprehensively described the metabolic strategies employed by V. dispar to thrive in the lactate-deficient environment using nitrate respiration and catabolism of specific amino acids. The elevated production of SCFAs and tryptophan from amino acids during nitrate respiration of V. dispar further suggested the potential roles of nitrate and Veillonella in the promotion of human health.
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Ácido Aspártico , Ácidos Graxos Voláteis , Ácido Glutâmico , Ácido Láctico , Nitratos , Triptofano , Veillonella , Triptofano/metabolismo , Ácidos Graxos Voláteis/metabolismo , Nitratos/metabolismo , Ácido Glutâmico/metabolismo , Ácido Aspártico/metabolismo , Veillonella/metabolismo , Veillonella/crescimento & desenvolvimento , Ácido Láctico/metabolismo , AnaerobioseRESUMO
The human gut hosts numerous ecological niches for microbe-microbe and host-microbe interactions. Gut lactate homeostasis in humans is crucial and relies on various bacteria. Veillonella spp., gut lactate-utilizing bacteria, and lactate-producing bacteria were frequently co-isolated. A recent clinical trial has revealed that lactate-producing bacteria in humans cross-feed lactate to Veillonella spp.; however, their interspecies interaction mechanisms remain unclear. Veillonella dispar, an obligate anaerobe commonly found in the human gut and oral cavity, ferments lactate into acetate and propionate. In our study, we investigated the interaction between V. dispar ATCC 17748T and three representative phylogenetically distant strains of lactic acid bacteria, Lactobacillus acidophilus ATCC 4356T, Lacticaseibacillus paracasei subsp. paracasei ATCC 27216T, and Lactiplantibacillus plantarum ATCC 10241. Bacterial growth, viability, metabolism and gene level adaptations during bacterial interaction were examined. V. dispar exhibited the highest degree of mutualism with L. acidophilus. During co-culture of V. dispar with L. acidophilus, both bacteria exhibited enhanced growth and increased viability. V. dispar demonstrated an upregulation of amino acid biosynthesis pathways and the aspartate catabolic pathway. L. acidophilus also showed a considerable number of upregulated genes related to growth and lactate fermentation. Our results support that V. dispar is able to enhance the fermentative capability of L. acidophilus by presumably consuming the produced lactate, and that L. acidophilus cross-feed not only lactate, but also glutamate, to V. dispar during co-culture. The cross-fed glutamate enters the central carbon metabolism in V. dispar. These findings highlight an intricate metabolic relationship characterized by cross-feeding of lactate and glutamate in parallel with considerable gene regulation within both L. acidophilus (lactate-producing) and V. dispar (lactate-utilizing). The mechanisms of mutualistic interactions between a traditional probiotic bacterium and a potential next-generation probiotic bacterium were elucidated in the production of short-chain fatty acids.
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Ácidos Graxos Voláteis , Ácido Glutâmico , Ácido Láctico , Veillonella , Ácido Láctico/metabolismo , Ácidos Graxos Voláteis/metabolismo , Ácido Glutâmico/metabolismo , Veillonella/metabolismo , Veillonella/crescimento & desenvolvimento , Veillonella/genética , Simbiose , Interações Microbianas , Humanos , Lactobacillus acidophilus/metabolismo , Lactobacillus acidophilus/crescimento & desenvolvimento , Lactobacillus acidophilus/genética , Lactobacillus/metabolismo , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Viabilidade Microbiana , FermentaçãoRESUMO
Here, we present the draft whole-genome sequence of Methylobacterium sp. OT2, isolated from human skin on a minimal medium containing t-octylphenol ethoxylate (Triton X-100). This genomic information contributes to understanding the niche adaptation on human skin and its catabolism of Triton X-100 in this strain.
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Veillonella spp. are obligate, anaerobic, Gram-negative bacteria found in the human oral cavity and gut. Recent studies have indicated that gut Veillonella promote human homeostasis by producing beneficial metabolites, specifically short-chain fatty acids (SCFAs), by lactate fermentation. The gut lumen is a dynamic environment with fluctuating nutrient levels, so the microbes present shifting growth rates and significant variations of gene expression. The current knowledge of lactate metabolism by Veillonella has focused on log phase growth. However, the gut microbes are mainly in the stationary phase. In this study, we investigated the transcriptomes and major metabolites of Veillonella dispar ATCC 17748T during growth from log to stationary phases with lactate as the main carbon source. Our results revealed that V. dispar reprogrammed its lactate metabolism during the stationary phase. Lactate catabolic activity and propionate production were significantly decreased during the early stationary phase but were partially restored during the stationary phase. The propionate/acetate production ratio was lowered from 1.5 during the log phase to 0.9 during the stationary phase. Pyruvate secretion was also greatly decreased during the stationary phase. Furthermore, we have demonstrated that the gene expression of V. dispar is reprogrammed during growth, as evidenced by the distinct transcriptomes present during the log, early stationary, and stationary phases. In particular, propionate metabolism (the propanediol pathway) was downregulated during the early stationary phase, which explains the decrease in propionate production during the stationary phase. The fluctuations in lactate fermentation during the stationary phase and the associated gene regulation expand our understanding of the metabolism of commensal anaerobes in changing environments. IMPORTANCE Short-chain fatty acids produced by gut commensal bacteria play an important role in human physiology. Gut Veillonella and the metabolites acetate and propionate, produced by lactate fermentation, are associated with human health. Most gut bacteria in humans are in the stationary phase. Lactate metabolism by Veillonella spp. during the stationary phase is poorly understood and was therefore the focus of the study. To this end, we used a commensal anaerobic bacterium and explored its short-chain fatty acid production and gene regulation in order to provide a better understanding of lactate metabolism dynamics during nutrient limitation.
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Rheumatoid arthritis (RA), an autoimmune disease, is characterized by chronic joint inflammation and pain. We previously found that the deletion of T-cell death-associated gene 8 (TDAG8) significantly reduces disease severity and pain in RA mice. Whether it is by modulating gut microbiota remains unclear. In this study, 64 intestinal samples of feces, cecal content, and cecal mucus from the complete Freund's adjuvant-induced arthritis mouse models were compared. The α- and ß-diversity indices of the microbiome were significantly lower in RA mice. Cecal mucus showed a higher ratio of Firmicutes to Bacteroidetes in RA than healthy mice, suggesting the ratio could serve as an RA indicator. Four core genera, Eubacterium_Ventriosum, Alloprevotella, Rikenella, and Treponema, were reduced in content in both feces and mucus RA samples, and could serve microbial markers representing RA progression. TDAG8 deficiency decreased the abundance of proinflammation-related Eubacterium_Xylanophilum, Clostridia, Ruminococcus, Paraprevotella, and Rikenellaceae, which reduced local mucosal inflammation to relieve RA disease severity and pain. The pharmacological block of the TDAG8 function by a salicylanilide derivative partly restored the RA microbiome to a healthy composition. These findings provide a further understanding of specific bacteria interactions with host gut mucus in the RA model. The modulation by TDAG8 on particular bacteria can facilitate microbiota-based therapy.
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Artrite Reumatoide , Microbioma Gastrointestinal , Microbiota , Animais , Bactérias/genética , Microbioma Gastrointestinal/genética , Inflamação , Camundongos , Dor , SalicilanilidasRESUMO
BACKGROUND/PURPOSE: The estimated prevalence of xerostomia (lack of saliva) ranges from 10% to 50% of the general population. The oral cavity provides a multivariant environmental habitat to over 700 species of bacteria and fungi. We hypothesized that xerostomia will alter the composition of oral microbiota. MATERIAL AND METHODS: Nineteen xerostomia patients and 10 healthy normal volunteers were studied for the oral microbiota. Gingival plaques were collected and microbiota were detected using bacterial 16S ribosomal RNA and analyzed based on the levels of phylum and class. RESULTS: In all cases, phyla of Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Proteobacteria make up to 100% of oral microbiota at phylum level. Analyzing individual phylum, presence of Bacteroidetes in xerostomia patients and normal subjects were 23.12 ± 2.56% and 23.23 ± 2.58%, respectively. Mean percentage presence of Firmicutes phylum in xerostomia patients and normal subjects were 18.94 ± 1.83% and 14.06 ± 0.98%, respectively. Statistically significant difference was not observed between xerostomia patients and normal subjects in this study. CONCLUSION: These observations revealed obvious but not statistically significant changes in oral major microorganism phylum between xerostomia patients and normal subjects in this study. More samples are needed to verify the current results and to use oral microbiota as a tool in the diagnosis of xerostomia.
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The shotgun genome sequence of Bacillus mycoides strain U53, a psychrotolerant bacterium that was isolated from soil in Russia, is characterized. The bacterial genome is 5,710,017 bp long and is predicted to have 5,847 genes. The GC content is 35.4%.
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BACKGROUND: The autoimmune disease rheumatoid arthritis (RA) affects approximately 1% of the global population. RA is characterized with chronic joint inflammation and often associated with chronic pain. The imbalance of pro-inflammatory and anti-inflammatory macrophages is a feature of RA progression. Glial cells affecting neuronal sensitivity at both peripheral and central levels may also be important for RA progression and associated pain. Genetic variants in the T cell death-associated gene 8 (TDAG8) locus are found to associate with spondyloarthritis. TDAG8 was also found involved in RA disease progression and associated hyperalgesia in the RA mouse model. However, its modulation in RA remains unclear. METHODS: To address this question, we intra-articularly injected complete Freund's adjuvant (CFA) into TDAG8+/+, TDAG8-/- or wild-type mice, followed by pain behavioral tests. Joints and dorsal root ganglia were taken, sectioned, and stained with antibodies to observe the number of immune cells, macrophages, and satellite glial cells (SGCs). For compound treatments, compounds were intraperitoneally or orally administered weekly for 9 consecutive weeks after CFA injection. RESULTS: We demonstrated that TDAG8 deletion slightly reduced RA pain in the early phase but dramatically attenuated RA progression and pain in the chronic phase (> 7 weeks). TDAG8 deletion inhibited an increase in SGC number and inhibition of SGC function attenuated chronic phase of RA pain, so TDAG8 could regulate SGC number to control chronic pain. TDAG8 deletion also reduced M1 pro-inflammatory macrophage number at 12 weeks, contributing to the attenuation of chronic RA pain. Such results were further confirmed by using salicylanilide derivatives, CCL-2d or LCC-09, to suppress TDAG8 expression and function. CONCLUSIONS: This study demonstrates that TDAG8 deletion reduced SGC and M1 macrophage number to relieve RA disease severity and associated chronic pain. M1 macrophages are critical for the development and maintenance of RA disease and pain, but glial activation is also required for the chronic phase of RA pain.
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Artrite Reumatoide/metabolismo , Macrófagos/imunologia , Neuroglia/imunologia , Animais , Artrite Experimental/imunologia , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Dor Crônica/imunologia , Dor Crônica/metabolismo , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Camundongos , Receptores Acoplados a Proteínas G/metabolismoRESUMO
4-t-octylphenol (4-t-OP), a well-known endocrine disrupting compound, is frequently found in various environmental compartments at levels that may cause adverse effects to the ecosystem and public health. To date, most of the studies that investigate microbial transformations of 4-t-OP have focused on the process mediated by bacteria, ligninolytic fungi, or microbial consortia. There is no report on the complete degradation mechanism of 4-t-OP by non-ligninolytic fungi. In this study, we conducted laboratory experiments to explore and characterize the non-ligninolytic fungal strain Fusarium falciforme RRK20 to degrade 4-t-OP. Using the response surface methodology, the initial biomass concentration and temperature were the factors identified to be more influential on the efficiency of the biodegradation process as compared with pH. Under the optimized conditions (i.e., 28⯰C, pH 6.5 with an initial inoculum density of 0.6â¯gâ¯L-1), 25â¯mgâ¯L-1 4-t-OP served as sole carbon source was completely depleted within a 14-d incubation; addition of low dosage of glucose was shown to significantly accelerate 4-t-OP degradation. The yeast estrogenic screening assay further confirmed the loss of estrogenic activity during the biodegradation process, though a longer incubation period was required for complete removal of estrogenicity. Metabolites identified by LC-MS/MS revealed that strain RRK20 might degrade 4-t-OP as sole energy source via alkyl chain oxidation and aromatic ring hydroxylation pathways. Together, these results not only suggest the potential use of non-ligninolytic fungi like strain RRK20 in remediation of 4-t-OP contaminated environments but may also improve our understanding of the environmental fate of 4-t-OP.
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Biodegradação Ambiental , Fusarium/química , Fenóis/química , Fungos/metabolismoRESUMO
Haematobacter massiliensis OT1 was isolated from human skin. This strain can catabolize 4-hydroxybenzoate. Here, we present the first complete whole-genome sequence of this species, which has one chromosome (2,467 kbp) and nine plasmids (total of 1,765 kbp). The analysis of the H. massiliensis OT1 genome indicated a potential for autotrophic growth.
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Bacteria in the environment play a major role in the degradation of widely used man-made recalcitrant organic compounds. Pseudomonas nitroreducens TX1 is of special interest because of its high efficiency to remove nonionic ethoxylated surfactants. In this study, a novel approach was demonstrated by a bacterial enzyme involved in the formation of radicals to attack ethoxylated surfactants. The dihydrolipoamide dehydrogenase was purified from the crude extract of strain TX1 by using octylphenol polyethoxylate (OPEOn) as substrate. The extent of removal of OPEOs during the degradation process was conducted by purified recombinant enzyme from E. coli BL21 (DE3) in the presence of the excess of metal mixtures (Mn2+, Mg2+, Zn2+, and Cu2+). The metabolites and the degradation rates were analyzed and determined by liquid chromatography-mass spectrometry. The enzyme was demonstrated to form Fenton reagent in the presence of an excess of metals. Under this in vitro condition, it was shown to be able to shorten the ethoxylate chains of OPEOn. After 2 hours of reaction, the products obtained from the degradation experiment revealed a prominent ion peak at m/z = 493.3, namely the ethoxylate chain unit is 6 (OPEO6) compared to OPEO9 (m/z = 625.3), the main undegraded surfactant in the no enzyme control. It revealed that the concentration of OPEO15 and OPEO9 decreased by 90% and 40% after 4 hours, respectively. The disappearance rates for the OPEOn homologs correlated to the length of the exothylate chains, suggesting it is not a specific enzymatic reaction which cleaves one unit by unit from the end of the ethoxylate chain. The results indicate the diverse and novel strategy by bacteria to catabolize organic compounds by using existing housekeeping enzyme(s).
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Proteínas de Bactérias/metabolismo , Di-Hidrolipoamida Desidrogenase/metabolismo , Fenóis/química , Fenóis/metabolismo , Pseudomonas/enzimologia , Tensoativos/química , Tensoativos/metabolismo , Cromatografia Líquida , Cobre/metabolismo , Escherichia coli/enzimologia , Magnésio/metabolismo , Manganês/metabolismo , Espectrometria de Massas , Zinco/metabolismoRESUMO
INTRODUCTION: Systemic lupus erythematosus (SLE) is a chronic and complex autoimmune disease with a wide range of clinical manifestations that affects multiple organs and tissues. Therefore the differential expression of proteins in the serum/plasma have potential clinical applications when treating SLE. METHODS: We have compared the plasma/serum protein expression patterns of nineteen active SLE patients with those of twelve age-matched and gender-matched healthy controls by proteomic analysis. To investigate the differentially expressed proteins among SLE and controls, a 2-dimensional gel electrophoresis coupled with high-resolution liquid chromatography tandem mass spectrometry was performed. To further understand the molecular and biological functions of the identified proteins, PANTHER and Gene Ontology (GO) analyses were employed. RESULTS: A total of 14 significantly expressed (p < 0.05, p < 0.01) proteins were identified, and of these nine were up-regulated and five down-regulated in the SLE patients. The functional enrichment analysis assigned the majority of the identified proteins including alpha 2 macroglobulin, complement C4, complement factor H, fibrinogen beta chain, and alpha-1-antitrypsin were part of the complement/coagulation cascade, which is an important pathway that plays a crucial role in SLE pathogenesis. In addition to these proteins the differential expressions of ceruloplasmin, transthyretin, and haptoglobin play a potential role in the renal system abnormalities of SLE. CONCLUSION: Therefore, the identified differentially expressed proteins are relevant to SLE patient's cohort. Most importantly the up-regulated proteins might be the potential candidates for renal system involvement in SLE disease pathogenesis. In order to confirm the diagnostic/therapeutic potential of the identified proteins, future validation studies are required.
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Biomarcadores/sangue , Proteínas Sanguíneas/metabolismo , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/metabolismo , Proteômica/métodos , Adulto , Estudos de Coortes , Feminino , Ontologia Genética , Humanos , Lúpus Eritematoso Sistêmico/diagnóstico , Masculino , Pessoa de Meia-Idade , alfa 1-Antitripsina/sangueRESUMO
CONTEXT: Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disease with unknown etiology. OBJECTIVE: Human plasma is comprised of over 10 orders of magnitude concentration of proteins and tissue leakages. The changes in the abundance of these proteins have played an important role in various human diseases. Therefore, the research objective of this study is to identify the significantly altered expression levels of plasma proteins from SLE patients compared with healthy controls using proteomic analysis. The plasma proteome profiles of both SLE patients and controls were compared. METHODS: A total of 19 active SLE patients and 12 healthy controls plasma samples were analyzed using high-resolution electrospray ionization liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) followed by label-free quantification. RESULTS: A total of 19 proteins showed a significant level of expression in the comparative LC-ESI-MS/MS triplicate analysis; among these, 14 proteins had >1.5- to three-fold up-regulation and five had <0.2- to 0.6-fold down-regulation. Gene ontology and DAVID (Database Annotation Visualization, and Integrated Discovery) functional enrichment analysis revealed that these proteins are involved in several important biological processes including acute phase inflammatory responses, complement activation, hemostasis, and immune system regulation. CONCLUSION: Our study identified a group of differentially expressed proteins in the plasma of SLE patients that are involved in the imbalance of the immune system and inflammatory responses. Therefore, these findings may have the potential to be used as prognostic/diagnostic markers for SLE disease assessment or disease monitoring.
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Here, we present the complete whole-genome sequences of three Moraxella osloensis strains with octylphenol polyethoxylate-degrading abilities. These strains were isolated from human skin.
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4-t-Octylphenol (4-t-OP) has become a serious environmental concern due to the endocrine disruption in animals and humans. The biodegradation of 4-t-OP by pure cultures has been extensively investigated only in bacteria and wood-decaying fungi. In this study we isolated and identified 14 filamentous fungal strains from wastewater samples in Taiwan using 4-t-OP as a sole carbon and energy source. The isolates were identified based on sequences from different DNA regions. Of 14 fungal isolates, 10 strains grew effectively on solid medium with a wide variety of endocrine disrupting chemicals as the sole carbon and energy source. As revealed by high-performance liquid chromatography analysis, the most effective 4-t-OP degradation (>70%) in liquid medium was observed in Fusarium falciforme after 15days. To our knowledge, this is the first report on the degradation of 4-t-OP as a sole carbon and energy source by non-ligninolytic fungi.
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Disruptores Endócrinos/metabolismo , Fungos/metabolismo , Fenóis/metabolismo , Poluentes Químicos da Água/metabolismo , Biodegradação Ambiental , Carbono/metabolismo , Cromatografia Líquida de Alta Pressão , Fungos/isolamento & purificação , Taiwan , Águas Residuárias/microbiologiaRESUMO
Rheumatoid arthritis (RA), characterized by chronic inflammation of synovial joints, is often associated with ongoing pain and increased pain sensitivity. High hydrogen ion concentration (acidosis) found in synovial fluid in RA patients is associated with disease severity. Acidosis signaling acting on proton-sensing receptors may contribute to inflammation and pain. Previous studies focused on the early phase of arthritis (<5 weeks) and used different arthritis models, so elucidating the roles of different proton-sensing receptors in the chronic phase of arthritis is difficult. We intra-articularly injected complete Freund's adjuvant into mice once a week for 4 weeks to establish chronic RA pain. Mice with knockout of acid-sensing ion channel 3 (ASIC3) or transient receptor potential/vanilloid receptor subtype 1 (TRPV1) showed attenuated chronic phase (>6 weeks) of RA pain. Mice with T-cell death-associated gene 8 (TDAG8) knockout showed attenuated acute and chronic phases of RA pain. TDAG8 likely participates in the initiation of RA pain, but all three genes, TDAG8, TRPV1, and ASIC3, are essential to establish hyperalgesic priming to regulate the chronic phase of RA pain.
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Canais Iônicos Sensíveis a Ácido/genética , Artrite Reumatoide/complicações , Artrite Reumatoide/genética , Hiperalgesia/etiologia , Hiperalgesia/fisiopatologia , Canais de Cátion TRPV/genética , Canais Iônicos Sensíveis a Ácido/metabolismo , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Artralgia/etiologia , Artralgia/fisiopatologia , Artrite Experimental , Artrite Reumatoide/patologia , Biomarcadores , Citocinas/sangue , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Expressão Gênica , Imuno-Histoquímica , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Camundongos , Camundongos Knockout , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Canais de Cátion TRPV/metabolismoRESUMO
4-(1,1,3,3-tetramethylbutane)-phenol (4-tert-OP) is one of the most prevalent endocrine disrupting pollutants. Information about bioremediation of 4-tert-OP remains limited, and no study has been reported on the mechanism of 4-tert-OP degradation by yeasts. The yeast Candida rugopelliculosa RRKY5 was proved to be able to utilize 4-methylphenol, bisphenol A, 4-ethylphenol, 4-tert-butylphenol, 4-tert-OP, 4-tert-nonylphenol, isooctane, and phenol under aerobic conditions. The optimum conditions for 4-tert-OP degradation were 30°C, pH 5.0, and an initial 4-tert-OP concentration of 30mgL-1; the maximum biodegradation rate constant was 0.107d-1, equivalent to a minimum half-life of 9.6d. Scanning electron microscopy revealed formation of arthroconidia when cells were grown in the presence of 4-tert-OP, whereas the cells remained in the budding form without 4-tert-OP. Identification of the 4-tert-OP degradation metabolites using liquid chromatography-hybrid mass spectrometry revealed three different mechanisms via both branched alkyl side chain and aromatic ring cleavage pathways.
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Candida/metabolismo , Disruptores Endócrinos/metabolismo , Fenóis/metabolismo , Biodegradação Ambiental , Cromatografia Líquida , Meia-Vida , Concentração de Íons de Hidrogênio , Hidroxilação , Espectrometria de Massas , Oxirredução , Fenóis/análiseRESUMO
Pseudomonas nitroreducens TX1 is of special interest because of its ability to utilize 0.05% to 20% octylphenol polyethoxylates (OPEOn) as a sole source of carbon. In this study, a library containing 30,000 Tn5-insertion mutants of the wild-type strain TX1 was constructed and screened for OPEOn utilization, and 93 mutants were found to be unable to grow on OPEOn In total, 42 separate disrupted genes were identified, and the proteins encoded by the genes were then classified into various categories, namely, information storage and processing (14.3%), cellular processes and signaling (28.6%), metabolism (35.7%), and unknown proteins (21.4%). The individual deletion of genes encoding isocitrate lyase (aceA), malate synthase (aceB), and glycolate dehydrogenase (glcE) was carried out, and the requirement for aceA and aceB but not glcE confirmed the role of the glyoxylate cycle in OPEOn degradation. Furthermore, acetaldehyde dehydrogenase and acetyl-coenzyme A (acetyl-CoA) synthetase activity levels were 13.2- and 2.1-fold higher in TX1 cells grown on OPEOn than in TX1 cells grown on succinate, respectively. Growth of the various mutants on different carbon sources was tested, and based on these findings, a mechanism involving exoscission to liberate acetaldehyde from the end of the OPEOn chain during degradation is proposed for the breakdown of OPEOn IMPORTANCE: Octylphenol polyethoxylates belong to the alkylphenol polyethoxylate (APEOn) nonionic surfactant family. Evidence based on the analysis of intermediate metabolites suggested that the primary biodegradation of APEOn can be achieved by two possible pathways for the stepwise removal of the C2 ethoxylate units from the end of the chain. However, direct evidence for these hypotheses is still lacking. In this study, we described the use of transposon mutagenesis to identify genes critical to the catabolism of OPEOn by P. nitroreducens TX1. The exoscission of the ethoxylate chain leading to the liberation of acetaldehyde is proposed. Isocitrate lyase and malate synthase in glyoxylate cycle are required in the catabolism of ethoxylated surfactants. Our findings also provide many gene candidates that may help elucidate the mechanisms in stress responses to ethoxylated surfactants by bacteria.
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Elementos de DNA Transponíveis , Mutagênese Insercional , Fenóis/metabolismo , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/genética , Acetaldeído/metabolismo , Oxirredutases do Álcool/genética , Oxirredutases do Álcool/metabolismo , Aldeído Oxirredutases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Deleção de Genes , Isocitrato Liase/genética , Isocitrato Liase/metabolismo , Malato Sintase/genética , Malato Sintase/metabolismo , Pseudomonas/metabolismo , Ácido Succínico/metabolismo , Tensoativos/metabolismoRESUMO
Bacterial degradation plays a vital role in determining the environmental fate of micropollutants like triclocarban. The mechanism of triclocarban degradation by pure bacterium is not yet explored. The purpose of this study was to identify metabolic pathway that might be involved in bacterial degradation of triclocarban. Triclosan-degrading Sphingomonas sp. strain YL-JM2C was first found to degrade up to 35% of triclocarban (4 mg L(-1)) within 5 d. Gas chromatography-mass spectrometry detected 3,4-dichloroaniline, 4-chloroaniline and 4-chlorocatechol as the major metabolites of the triclocarban degradation. Furthermore, total organic carbon results confirmed that the intermediates, 3,4-dichloroaniline (4 mg L(-1)) and 4-chloroaniline (4 mg L(-1)) could be degraded up to 77% and 80% by strain YL-JM2C within 5 d.
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Carbanilidas/análise , Poluentes Ambientais/análise , Sphingomonas/crescimento & desenvolvimento , Triclosan/análise , Compostos de Anilina/análise , Compostos de Anilina/metabolismo , Biodegradação Ambiental , Carbanilidas/metabolismo , Catecóis/análise , Catecóis/metabolismo , China , Poluentes Ambientais/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Sphingomonas/metabolismo , Triclosan/metabolismoRESUMO
BACKGROUND: Long-chain alkylphenols, such as nonylphenols and octylphenols, are man-made compounds with estrogen-like activity and considered to be endocrine disputing chemicals. They are known to influence sexual development and reproduction of humans and other animals, therefore cause health and ecological concerns. OBJECTIVE: This review aims to identify the key findings presented in recent scientific literature with respect to the presence of long-chain alkylphenols in food, intake by different populations and describe the metabolism by human and the mechanisms to disrupt the estrogenic activity of nonylphenols and octylphenols by bacteria and fungi in the environment. METHOD: The current knowledge related to the physicochemical properties, source, occurrence, toxic effects, level in foods, human intake, elimination and metabolism of long-chain alkylphenols was summarized. RESULTS: Long-chain alkylphenols have been detected in soil, surface and ground water, sediments, wastewater effluent, air and in most foods. The daily human intake of 4-nonylphenols and 4-octylphenols has been compared among different population. A comprehensive summary of the major degradation pathways and mechanism in bacteria, fungi, yeasts and vertebrates was described. CONCLUSION: The findings of this review revealed the wide distribution of long-chain alkylphenols in various environments and contamination in foods. Therefore, further investigation of long-term exposure is necessary to comprehensively evaluate the potential health risks to humans. The diverse biodegradation mechanisms can be used in the removal of these compounds before entering food chains. The review provides indication as to how it might be possible to reduce the adverse effects on human health from estrogen-like long-chain alkylphenols.