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1.
Microbiol Res ; 286: 127806, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38924817

RESUMO

Metabolic cross-feeding is a pervasive interaction between bacteria to acquire novel phenotypes. However, our current understanding of the survival mechanism for cross-feeding in cocultured bacterial biofilms under heavy-metal conditions remains limited. Herein, we found that Comamonas sp. A23 produces L-phenylalanine to activate the L-phenylalanine degradation pathway in Enterobacter sp. A11, enhancing biofilm formation and cadmium [Cd(II)] immobilization in A11. The genes responsible for L-phenylalanine-degradation (paaK) and cell attachment and aggregation (csgAD) are essential for biofilm formation and Cd(II) immobilization in A11 induced by L-phenylalanine. The augmentation of A11 biofilms, in turn, protects A23 under Cd(II) and H2O2 stresses. The plant-based experiments demonstrate that the induction of two rice Cd(II) transporters, OsCOPT4 and OsBCP1, by A11 and A23 enhances rice resistance against Cd(II) and H2O2 stresses. Overall, our findings unveil the mutual dependence between bacteria and rice on L-phenylalanine cross-feeding for survival under abiotic stress.


Assuntos
Biofilmes , Cádmio , Comamonas , Enterobacter , Peróxido de Hidrogênio , Oryza , Fenilalanina , Cádmio/metabolismo , Oryza/microbiologia , Enterobacter/metabolismo , Enterobacter/genética , Biofilmes/crescimento & desenvolvimento , Peróxido de Hidrogênio/metabolismo , Fenilalanina/metabolismo , Comamonas/metabolismo , Comamonas/genética , Estresse Fisiológico , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Interações Microbianas
2.
Artigo em Inglês | MEDLINE | ID: mdl-38190241

RESUMO

Five strains of two novel species were isolated from the wastewater treatment systems of a pharmaceutical factory located in Zhejiang province, PR China. Strains ZM22T and Y6 were identified as belonging to a potential novel species of the genus Comamonas, whereas strains ZM23T, ZM24 and ZM25 were identified as belonging to a novel species of the genus Pseudomonas. These strains were characterized by polyphasic approaches including 16S rRNA gene analysis, multi-locus sequence analysis, average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH), physiological and biochemical tests, as well as chemotaxonomic analysis. Genome-based phylogenetic analysis further confirmed that strains ZM22T and Y6 form a distinct clade closely related to Comamonas testosteroni ATCC 11996T and Comamonas thiooxydans DSM 17888T. Strains ZM23T, ZM24 and ZM25 were grouped as a separate clade closely related to Pseudomonas nitroreducens DSM 14399T and Pseudomonas nicosulfuronedens LAM1902T. The orthoANI and isDDH results indicated that strains ZM22T and Y6 belong to the same species. In addition, genomic DNA fingerprinting demonstrated that these strains do not originate from a single clone. The same results were observed for strains ZM23T, ZM24 and ZM25. Strains ZM22T and Y6 were resistant to multiple antibiotics, whereas strains ZM23T, ZM24 and ZM25 were able to degrade an emerging pollutant, triclosan. The phylogenetic, physiological and biochemical characteristics, as well as chemotaxonomy, allowed these strains to be distinguished from their genus, and we therefore propose the names Comamonas resistens sp. nov. (type strain ZM22=MCCC 1K08496T=KCTC 82561T) and Pseudomonas triclosanedens sp. nov. (type strain ZM23T=MCCC 1K08497T=JCM 36056T), respectively.


Assuntos
Comamonas , Ácidos Graxos , Purificação da Água , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonas/genética , DNA Bacteriano/genética , Ácidos Graxos/química , Filogenia , Pseudomonas/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Indústria Farmacêutica
3.
Artigo em Inglês | MEDLINE | ID: mdl-38190243

RESUMO

Two novel indole acetic acid-producing strains, 5MLIRT and D4N7, were isolated from Indosasa shibataeoides in Yongzhou, Hunan province, and Phyllostachys edulis in Hangzhou, Zhejiang province, respectively. Based on their 16S rRNA sequences, strains 5MLIRT and D4N7 were closely related to Comamonas antarcticus 16-35-5T (98.4 % sequence similarity), and the results of 92-core gene phylogenetic trees showed that strains 5MLIRT and D4N7 formed a phylogenetic lineage within the clade comprising Comamonas species. The complete genome size of strain 5MLIRT was 4.49 Mb including two plasmids, and the DNA G+C content was 66.5 mol%. The draft genome of strain D4N7 was 4.26 Mb with 66.7 mol% G+C content. The average nucleotide identity and digital DNA-DNA hybridization values among strain 5MLIRT and species in the genus Comamonas were all below the species delineation threshold. The colonies of strain 5MLIRT and D4N7 were circular with regular margins, convex, pale yellow and 1.0-2.0 mm in diameter when incubated at 30 °C for 3 days. Strains 5MLIRT and D4N7 grew optimally at 30 °C, pH 7.0 and 1.0 % NaCl. The respiratory isoprenoid quinone was ubiquinone-8. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Polyphasic analyses indicated that strains 5MLIRT and D4N7 could be distinguished from related validly named Comamonas species and represent a novel species of the genus Comamonas, for which the name Comamonas endophytica sp. nov. is proposed. The type strain is 5MLIRT (=ACCC 62069T=GDMCC 1.2958T=JCM 35331T).


Assuntos
Comamonas , Endófitos , Composição de Bases , Endófitos/genética , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Ácidos Graxos/química , China , Poaceae
4.
Mol Plant Microbe Interact ; 36(2): 95-108, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36366828

RESUMO

The accumulation of cadmium (Cd) in plants is strongly impacted by soil microbes, but its mechanism remains poorly understood. Here, we report the mechanism of reduced Cd accumulation in rice by coculture of Enterobacter and Comamonas species. In pot experiments, inoculation with the coculture decreased Cd content in rice grain and increased the amount of nonbioavailable Cd in Cd-spiked soils. Fluorescence in situ hybridization and scanning electron microscopy detection showed that the coculture colonized in the rhizosphere and rice root vascular tissue and intercellular space. Soil metagenomics data showed that the coculture increased the abundance of sulfate reduction and biofilm formation genes and related bacterial species. Moreover, the coculture increased the content of organic matter, available nitrogen, and potassium and increased the activities of arylsulfatase, ß-galactosidase, phenoloxidase, arylamidase, urease, dehydrogenase, and peroxidase in soils. In subsequent rice transcriptomics assays, we found that the inoculation with coculture activated a hypersensitive response, defense-related induction, and mitogen-activated protein kinase signaling pathway in rice. Heterologous protein expression in yeast confirmed the function of four Cd-binding proteins (HIP28-1, HIP28-4, BCP2, and CID8), a Cd efflux protein (BCP1), and three Cd uptake proteins (COPT4, NRAM5, and HKT6) in rice. Succinic acid and phenylalanine were subsequently proved to inhibit rice divalent Cd [Cd(II)] uptake and activate Cd(II) efflux in rice roots. Thus, we propose a model that the coculture protects rice against Cd stress via Cd immobilization in soils and reducing Cd uptake in rice. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Assuntos
Comamonas , Oryza , Poluentes do Solo , Cádmio/análise , Cádmio/metabolismo , Oryza/metabolismo , Enterobacter/genética , Comamonas/metabolismo , Técnicas de Cocultura , Hibridização in Situ Fluorescente , Solo/química , Poluentes do Solo/análise , Poluentes do Solo/metabolismo
5.
Artigo em Inglês | MEDLINE | ID: mdl-35316175

RESUMO

A Gram-stain-negative, aerobic and motile bacterial strain, designated CJ34T, was isolated from Han River water in the Republic of Korea. Strain CJ34T grew optimally on tryptic soy agar at 30 °C and pH 7.0 in the absence of NaCl. Results of phylogenetic analysis based on 16S rRNA gene sequence showed that strain CJ34T belonged to the genus Comamonas within the family Comamonadaceae and was most closely related to Comamonas testosteroni ATCC 11996T and Comamonas thiooxydans DSM 17888T (both 98.63 % similarity). The average nucleotide identity values between strain CJ34T and two closely related type strains C. testosteroni ATCC 11996T and C. thiooxydans DSM 17888T were 82.77 and 82.73 %, respectively. The major isoprenoid quinone of strain CJ34T was ubiquinone Q-8. The major cellular fatty acids of strain CJ34T were C16 : 0, C16 : 1 ω6c and/or C16 : 1 ω7c and C18 : 1 ω6c and/or C18 : 1 ω7c. The predominant polar lipids of strain CJ34T were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified aminophospholipid. Whole genome sequencing revealed that strain CJ34T had a genome of 4.9 Mbp and the G+C content of the genomic DNA was 59.73 mol%. On the basis of the results of this polyphasic taxonomy study, strain CJ34T represents a novel species in the genus Comamonas, for which the name Comamonas fluminis sp. nov. is proposed. The type strain is CJ34T (=KACC 22237T=JCM 34454T).


Assuntos
Comamonas , Rios , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonas/genética , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Rios/microbiologia , Análise de Sequência de DNA
6.
Artigo em Inglês | MEDLINE | ID: mdl-33847556

RESUMO

A novel bacterial strain, EJ-4T, isolated from stream water collected at Seo-ho in Suwon, Republic of Korea, was characterized based on a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain EJ-4T belonged to the genus Comamonas. The isolate is Gram-stain-negative, non-motile, aerobic, rod-shaped and forms pale yellow colonies on trypticase soy agar. The optimal growth of this strain was observed aerobically at 30 °C, pH 7 and 0.5 % NaCl. The major fatty acids were summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c; 39.7 %) and C16 : 0 (32.0 %). The G+C content of strain EJ-4T was 58.4mol %. The average nucleotide identity and digital DNA-DNA hybridization values between strain EJ-4T and Comamoas testosteroni were 91.8 and 31.2 %, respectively. The major polar lipids detected in the isolate were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The predominant isoprenoid quinone was ubiquinone-8. Based on the results of polyphasic taxonomic analysis of strain EJ-4T, we describe a novel species of the genus Comamonas, for which the name Comamonas suwonensis sp. nov. has been proposed, with EJ-4T (=KCTC 82074T=JCM 34179T=KEMB 1602-279T) as the type strain.


Assuntos
Comamonas/classificação , Filogenia , Rios/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonas/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/química
7.
J Hazard Mater ; 403: 123824, 2021 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-33264916

RESUMO

Cadmium (Cd) is a typical heavy-metal highly accumulating in crops and drinking water, thus posing a severe health risk for human health. In this study, we firstly isolated 24 Cd-resistant bacteria from the heavy-metals contaminated soil at Daye Iron Ore, in which Comamonas sp. XL8 exhibited a high resistance and strong accumulation capacity to Cd. After absorption, Comamonas sp. XL8 could biosynthesize intracellular Cd-nanoparticles (CdNPs), which has not been reported in characteristics of Comamonas genus before. We found that the gene expressions of cadA and bmtA related to Cd transportation and binding in strain XL8 were significantly upregulated with Cd exposure, suggesting that genes cadA and bmtA may contribute to the formation of CdNPs. Of particular note, the co-inoculation of Comamonas sp. XL8 and rice seedlings (Oryzae sativa L.) significantly decreased the oxidative stress-induced by Cd accumulation and subsequently alleviated toxicity of Cd exposure. Our results reveal the biochemical process of Cd accumulation in Comamonas sp. XL8 by forming CdNPs, showing that it has great potential for effective bioremediation of environmental Cd exposure.


Assuntos
Comamonas , Oryza , Poluentes do Solo , Bactérias , Biodegradação Ambiental , Cádmio/análise , Cádmio/toxicidade , Humanos , Plântula , Solo , Poluentes do Solo/análise , Poluentes do Solo/toxicidade
8.
Rev. chil. infectol ; 37(2): 179-181, abr. 2020.
Artigo em Espanhol | LILACS | ID: biblio-1126106

RESUMO

Resumen Comunicamos la primera identificación en Ecuador del microorganismo Comamonas kerstersii, cepa aislada a partir de una muestra de líquido de la cavidad abdominal e identificada mediante la técnica de espectrometría de masas MALDITOF. Fue obtenida durante el acto quirúrgico de un paciente con el diagnóstico de una peritonitis aguda, secundaria a una apendicitis perforada. Este microorganismo es considerado un patógeno poco común. Pese a que el género al que pertenece tiene una amplia distribución geográfica, es considerado un agente oportunista.


Abstract We report the first identification in Ecuador of the microorganism Comamonas kerstersii, a strain isolates from a sample of fluid from the abdominal cavity and identified by the technique of mass spectrometry MALDITOF. It was obtained during the surgical act of a patient with the diagnosis of acute peritonitis, secondary to a perforated appendicitis. This microorganism is considered a rare pathogen. Even though the genus to which it belongs has a wide geographical distribution, this pathogen is considered an opportunistic agent.


Assuntos
Humanos , Comamonas , Peritonite , Equador
9.
Rev. chil. infectol ; 37(2): 182-185, abr. 2020. tab
Artigo em Espanhol | LILACS, UY-BNMED, BNUY | ID: biblio-1126107

RESUMO

Resumen Comamonas kerstersii es un bacilo gramnegativo no fermentador, asociado a infecciones intraabdominales. Comunicamos un caso clínico de una bacteriemia por C. kerstersii en un joven con una apendicitis aguda. La primera identificación, mediante el sistema Vitek2 compact (BioMerieux) desde los hemocultivos, fue Comamonas testosteroni. Sin embargo, mediante MALDI-TOF se identificó como C. kerstersii. El paciente fue tratado con ampicilina/sulbactam y luego piperacilina/tazobactam con una evolución favorable. Dentro del género Comamonas, C. testosteroni ha sido la especie más frecuentemente reportada como causa de infecciones en humanos, describiéndose solo 23 casos de C. kerstersii. Dado los problemas en la identificación fenotípica de estos microorganismos, es posible que algunos de los reportes de C. testosteroni pudieran haber correspondido a C. kerstersii. Este caso pone en evidencia el rol patógeno de C. kerstersii y la importancia de utilizar MALDI-TOF como herramienta diagnóstica en bacilos gramnegativos no fermentadores.


Abstract Comamonas kerstersii is a non-fermenting Gram-negative bacillus. It has been associated with intra-abdominal infections. We describe a clinical case of bacteremia caused by C. kerstersii in a young man with acute appendicitis. The first identification, using the Vitek2 compact system (BioMerieux) from blood cultures, was Comamonas testosteroni. However, using MALDI-TOF was identified as C. kerstersii. The patient was treated first with ampicillin sulbactam and then piperacillin tazobactam with favorable evolution. Within the genus Comamonas, C. testosteroni has been the most frequently reported species as a cause of infections in humans, only 23 cases of C. kerstersii being described. Given the problems of phenotypic identification of these microorganisms, it is possible that some C. testosteroni reports could have corresponded to C. kerstersii. This case highlights the pathogenic role of C. kerstersii and the importance of using MALDI-TOF as a diagnostic tool for non-fermenting Gram-negative bacilli identification.


Assuntos
Humanos , Masculino , Apendicite , Bacteriemia , Comamonas , Bactérias Gram-Negativas
10.
Curr Microbiol ; 76(4): 470-477, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30783796

RESUMO

Several studies indicated that Odorrana grahami (O. grahami) skin contains abundant antimicrobial peptides, and the skin was recognized as hostile habitat for microorganisms. In this study, the microbial community was evaluated by 16S rRNA gene sequencing, and two associated bacterial isolates were obtained and characterized from the skin of O. grahami. Sixteen bacterial genera were identified from the O. grahami skin by uncultured clone sequences. The dominant groups were Comamonas, Bacillus and Morganella, and the genus Comamonas was the most abundant group (41.7% of the total) of the community. Fortunately, strains CW-25T and CW-518 belonging to genus Comamonas were isolated by plating dilutions. The polyphasic taxonomy results indicated that strain CW-25T was a member of Comamonas aquatica, it showed much higher antimicrobials resistance than the closest C. aquatica strains of LMG 2370T, LMG5937 and LMG 6112 isolated from freshwater. Based on the polyphasic taxonomic studies and antimicrobials resistance characteristics, two subspecies of Comamonas aquatica subsp. aquatica nov. and Comamonas aquatica subsp. rana nov. were proposed. The super-antimicrobial resistance endows the strains of Comamonas aquatica subsp. rana inhabit the O. grahami skin, and the primary defense of O. grahami might be composed by the antimicrobial peptides and the native bacteria.


Assuntos
Bactérias/classificação , Comamonas/classificação , Comamonas/fisiologia , Filogenia , Ranidae/microbiologia , Pele/microbiologia , Animais , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Bactérias/genética , Comamonas/efeitos dos fármacos , Comamonas/genética , DNA Bacteriano/genética , Farmacorresistência Bacteriana Múltipla , Testes de Sensibilidade Microbiana , Hibridização de Ácido Nucleico , Fenótipo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Pele/química
11.
Cell ; 169(3): 431-441.e8, 2017 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-28431244

RESUMO

The human microbiota greatly affects physiology and disease; however, the contribution of bacteria to the response to chemotherapeutic drugs remains poorly understood. Caenorhabditis elegans and its bacterial diet provide a powerful system to study host-bacteria interactions. Here, we use this system to study how bacteria affect the C. elegans response to chemotherapeutics. We find that different bacterial species can increase the response to one drug yet decrease the effect of another. We perform genetic screens in two bacterial species using three chemotherapeutic drugs: 5-fluorouracil (5-FU), 5-fluoro-2'-deoxyuridine (FUDR), and camptothecin (CPT). We find numerous bacterial nucleotide metabolism genes that affect drug efficacy in C. elegans. Surprisingly, we find that 5-FU and FUDR act through bacterial ribonucleotide metabolism to elicit their cytotoxic effects in C. elegans rather than by thymineless death or DNA damage. Our study provides a blueprint for characterizing the role of bacteria in the host response to chemotherapeutics.


Assuntos
Antineoplásicos/metabolismo , Caenorhabditis elegans/microbiologia , Comamonas/metabolismo , Escherichia coli/metabolismo , Microbioma Gastrointestinal , Animais , Antineoplásicos/farmacologia , Camptotecina/metabolismo , Camptotecina/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Comamonas/genética , Desoxiuridina/análogos & derivados , Desoxiuridina/metabolismo , Desoxiuridina/farmacologia , Dieta , Escherichia coli/genética , Fluoruracila/metabolismo , Fluoruracila/farmacologia , Humanos , Modelos Animais , Nucleosídeos de Pirimidina/metabolismo
12.
Appl Microbiol Biotechnol ; 101(12): 5163-5173, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28299401

RESUMO

The surfactant sodium lauryl ether sulfate (SLES) is widely used in the composition of detergents and frequently ends up in wastewater treatment plants (WWTPs). While aerobic SLES degradation is well studied, little is known about the fate of this compound in anoxic environments, such as denitrification tanks of WWTPs, nor about the bacteria involved in the anoxic biodegradation. Here, we used SLES as sole carbon and energy source, at concentrations ranging from 50 to 1000 mg L-1, to enrich and isolate nitrate-reducing bacteria from activated sludge of a WWTP with the anaerobic-anoxic-oxic (A2/O) concept. In the 50 mg L-1 enrichment, Comamonas (50%), Pseudomonas (24%), and Alicycliphilus (12%) were present at higher relative abundance, while Pseudomonas (53%) became dominant in the 1000 mg L-1 enrichment. Aeromonas hydrophila strain S7, Pseudomonas stutzeri strain S8, and Pseudomonas nitroreducens strain S11 were isolated from the enriched cultures. Under denitrifying conditions, strains S8 and S11 degraded 500 mg L-1 SLES in less than 1 day, while strain S7 required more than 6 days. Strains S8 and S11 also showed a remarkable resistance to SLES, being able to grow and reduce nitrate with SLES concentrations up to 40 g L-1. Strain S11 turned out to be the best anoxic SLES degrader, degrading up to 41% of 500 mg L-1. The comparison between SLES anoxic and oxic degradation by strain S11 revealed differences in SLES cleavage, degradation, and sulfate accumulation; both ester and ether cleavage were probably employed in SLES anoxic degradation by strain S11.


Assuntos
Desnitrificação , Bactérias Gram-Negativas/metabolismo , Dodecilsulfato de Sódio/análogos & derivados , Aeromonas/isolamento & purificação , Aeromonas/metabolismo , Biodegradação Ambiental , Carbono/metabolismo , Comamonadaceae/isolamento & purificação , Comamonadaceae/metabolismo , Comamonas/isolamento & purificação , Comamonas/metabolismo , Bactérias Gram-Negativas/genética , Bactérias Gram-Negativas/isolamento & purificação , Oxirredução , Pseudomonas/isolamento & purificação , Pseudomonas/metabolismo , Esgotos/microbiologia , Dodecilsulfato de Sódio/química , Dodecilsulfato de Sódio/metabolismo , Tensoativos/química , Tensoativos/metabolismo
13.
Bioresour Technol ; 216: 684-90, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27289060

RESUMO

Magnetically immobilized cells of Comamonas sp. JB coupling with electrode reaction was developed to enhance the treatment efficiency of coking wastewater containing phenol, carbazole (CA), dibenzofuran (DBF), and dibenzothiophene (DBT). The pair of graphite plate-stainless iron mesh electrodes was chosen as the most suitable electrodes. Magnetically immobilized cells coupling with graphite plate-stainless iron mesh electrodes (coupling system) exhibited high degradation activity for all the compounds, which were significantly higher than the sum by single magnetically immobilized cells and electrode reaction at the optimal voltage. Recycling experiments demonstrated that the degradation activity of coupling system increased gradually during eight recycles, indicating that there was a coupling effect between the biodegradation and electrode reaction. Phenol hydroxylase and qPCR assays confirmed that appropriate electrical stimulation could improve phenol hydroxylase activity and promote cells growth. Toxicity assessment suggested the treatment of the coking wastewater by coupling system led to less toxicity than untreated wastewater.


Assuntos
Coque , Comamonas/química , Eliminação de Resíduos Líquidos/métodos , Águas Residuárias/química , Benzofuranos/química , Benzofuranos/metabolismo , Biodegradação Ambiental , Carbazóis/química , Carbazóis/metabolismo , Células Imobilizadas/química , Comamonas/metabolismo , Eletrodos , Grafite , Ferro , Magnetismo , Oxigenases de Função Mista/química , Oxigenases de Função Mista/metabolismo , Fenol/química , Fenol/metabolismo , Tiofenos/química , Tiofenos/metabolismo , Eliminação de Resíduos Líquidos/instrumentação
14.
J Appl Microbiol ; 120(6): 1542-51, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26918381

RESUMO

AIMS: To improve the efficiency of asymmetric hydrolysis of 3-(4-chlorophenyl) glutaric acid diamide (CGD) using a recombinant Comamonas sp. KNK3-7 amidase (CoAM) produced in Escherichia coli. METHODS AND RESULTS: The CoAM gene was cloned, sequenced and found to comprise 1512 bp, encoding a polypeptide of 54 054 Da. CoAM-transformed E. coli were able to perform R-selective hydrolysis of CGD; however, complete conversion of 166·2 mmol l(-1) CGD in 28 h could not be obtained. We attempted to optimize the reactivity of CoAM by mutating single amino acids in the substrate-binding domain. Notably, the methionine-substituted L146M mutant enzyme showed increased reactivity, completing the conversion of 166·2 mmol l(-1) CGD in just 4 h. The Km value for L146M was lower than that of CoAM. CONCLUSIONS: We succeeded in creating the L146M mutant of CoAM with increased substrate affinity and found that this was the best mutant for the hydrolysis of CGD. SIGNIFICANCE AND IMPACT OF THE STUDY: Increasing the efficiency of hydrolysis of 3-substituted glutaric acid diamides is useful to improve the synthesis of optically active 3-substituted gamma-aminobutyric acid. This is the first report of efficient hydrolysis of CGD using amidase mutant-producing E. coli cells.


Assuntos
Amidoidrolases/genética , Comamonas/enzimologia , Comamonas/genética , Diamida/química , Glutaratos/química , Engenharia de Proteínas , Amidoidrolases/química , Amidoidrolases/isolamento & purificação , Sítios de Ligação , Clonagem Molecular , Comamonas/metabolismo , Escherichia coli/genética , Hidrólise , Reação em Cadeia da Polimerase , Rhodococcus/enzimologia
15.
Appl Biochem Biotechnol ; 176(6): 1700-8, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26018344

RESUMO

A bacterium designated strain JB, able to degrade six benzene, toluene, ethylbenzene, and o-, m-, and p-xylene (BTEX) compounds, was isolated from petroleum-contaminated soil. Taxonomic analyses showed that the isolate belonged to Comamonas, and until now, the genus Comamonas has not included any known BTEX degraders. The BTEX biodegradation rate was slightly low on the mineral salt medium (MSM), but adding a small amount of yeast extract greatly enhanced the biodegradation. The relationship between specific degradation rate and individual BTEX was described well by Michaelis-Menten kinetics. The treatment of petrochemical wastewater containing BTEX mixture and phenol was shown to be highly efficient by BTEX-grown JB. In addition, toxicity assessment indicated the treatment of the petrochemical wastewater by BTEX-grown JB led to less toxicity than untreated wastewater.


Assuntos
Derivados de Benzeno/metabolismo , Benzeno/metabolismo , Comamonas/isolamento & purificação , Comamonas/metabolismo , Tolueno/metabolismo , Xilenos/metabolismo
16.
Int J Syst Evol Microbiol ; 64(Pt 12): 3976-3982, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25212224

RESUMO

A bacterial strain, designated GAU11(T), was isolated from soil in Japan. Cells of the strain were Gram-stain-negative, aerobic, non-motile rods. The 16S rRNA gene sequence of strain GAU11(T) showed high similarity to those of Comamonas zonglianii BF-3(T) (98.8 %), Pseudacidovorax intermedius CC21(T) (96.4 %), Acidovorax caeni R-24608(T) (96.2 %), Alicycliphilus denitrificans K601(T) (96.2 %), Pseudorhodoferax soli TBEA3(T) (95.9 %) and Comamonas terrigena LMG 1253(T) (95.9 %). Strain GAU11(T) contained ubiquinone 8 as the sole ubiquinone and diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol as major polar lipids. Its major cellular fatty acids were C16 : 0, C18 : 1ω7c and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The DNA G+C content of strain GAU11(T) was 68.2 mol%. The DNA-DNA relatedness between strain GAU11(T) and C. zonglianii DSM 22523(T) was 52 or 68 % (reciprocal value). Phenotypic characterization indicated that strain GAU11(T) represents a member of the genus Comamonas, but at the same time distinguished it from C. zonglianii DSM 22523(T). From polyphasic characterization, this strain should be classified as representing a novel species of the genus Comamonas, for which the name Comamonas humi sp. nov. (type strain GAU11(T) = JCM 19903(T) = DSM 28451(T)) is proposed.


Assuntos
Comamonas/classificação , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonas/genética , Comamonas/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Japão , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química
17.
Proc Natl Acad Sci U S A ; 111(18): 6702-7, 2014 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-24733922

RESUMO

Aerolysins are virulence factors belonging to the bacterial ß-pore-forming toxin superfamily. Surprisingly, numerous aerolysin-like proteins exist in vertebrates, but their biological functions are unknown. ßγ-CAT, a complex of an aerolysin-like protein subunit (two ßγ-crystallin domains followed by an aerolysin pore-forming domain) and two trefoil factor subunits, has been identified in frogs (Bombina maxima) skin secretions. Here, we report the rich expression of this protein, in the frog blood and immune-related tissues, and the induction of its presence in peritoneal lavage by bacterial challenge. This phenomena raises the possibility of its involvement in antimicrobial infection. When ßγ-CAT was administrated in a peritoneal infection model, it greatly accelerated bacterial clearance and increased the survival rate of both frogs and mice. Meanwhile, accelerated Interleukin-1ß release and enhanced local leukocyte recruitments were determined, which may partially explain the robust and effective antimicrobial responses observed. The release of interleukin-1ß was potently triggered by ßγ-CAT from the frog peritoneal cells and murine macrophages in vitro. ßγ-CAT was rapidly endocytosed and translocated to lysosomes, where it formed high molecular mass SDS-stable oligomers (>170 kDa). Lysosomal destabilization and cathepsin B release were detected, which may explain the activation of caspase-1 inflammasome and subsequent interleukin-1ß maturation and release. To our knowledge, these results provide the first functional evidence of the ability of a host-derived aerolysin-like protein to counter microbial infection by eliciting rapid and effective host innate immune responses. The findings will also largely help to elucidate the possible involvement and action mechanisms of aerolysin-like proteins and/or trefoil factors widely existing in vertebrates in the host defense against pathogens.


Assuntos
Interações Hospedeiro-Patógeno/imunologia , Peptídeos/imunologia , Proteínas Citotóxicas Formadoras de Poros/imunologia , Sequência de Aminoácidos , Animais , Anuros/genética , Anuros/imunologia , Anuros/microbiologia , Toxinas Bacterianas/química , Toxinas Bacterianas/genética , Toxinas Bacterianas/imunologia , Comamonas , Endocitose , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/prevenção & controle , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Imunidade Inata , Inflamassomos/imunologia , Interleucina-1beta/biossíntese , Lisossomos/imunologia , Masculino , Camundongos , Modelos Moleculares , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/genética , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/genética , Conformação Proteica , Homologia de Sequência de Aminoácidos , Pele/imunologia , Pele/microbiologia , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/prevenção & controle , Fator Trefoil-2
18.
Int J Syst Evol Microbiol ; 63(Pt 3): 809-814, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22581903

RESUMO

A facultatively anaerobic bacterium, strain CY01(T), isolated from subterranean forest sediment collected from Guangdong Province, China, was investigated using a polyphasic taxonomic approach. The cells were short rods, Gram-negative, non-sporulating and motile. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CY01(T) showed highest sequence similarities to Comamonas thiooxydans S23(T) (98.0 %), Comamonas testosteroni JCM 5832(T) (97.9 %), Comamonas koreensis KCTC 12005(T) (97.7 %) and Comamonas odontotermitis LMG 23579(T) (97.0 %). The major respiratory quinone was ubiquinone-8. The major cellular fatty acids were summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), C16 : 0 and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). Based on the phylogenetic analysis, DNA-DNA hybridization, whole-cell fatty acid composition as well as biochemical characteristics, strain CY01(T) was clearly distinguishable from all recognized species of the genus Comamonas and should be classified as a representative of a novel species of the genus, for which the name Comamonas guangdongensis sp. nov. is proposed. The type strain is CY01(T) ( = CCTCC AB 2011133(T) = KACC 16241(T)).


Assuntos
Comamonas/classificação , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Comamonas/genética , Comamonas/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/análise , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Árvores/microbiologia , Ubiquinona/análise
19.
Int J Syst Evol Microbiol ; 63(Pt 6): 2168-2173, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23125317

RESUMO

A novel biosurfactant-producing strain, designated YW1(T), was isolated from agricultural soil. Its taxonomic position was investigated using a polyphasic approach. The cells were short rods, Gram-negative, non-sporulating and motile. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YW1(T) was a member of the genus Comamonas, and showed highest sequence similarities to Comamonas aquatica LMG 2370(T) (98.5%), Comamonas kerstersii LMG 3475(T) (97.7%) and Comamonas terrigena LMG 1253(T) (97.7%). Furthermore, DNA-DNA hybridization experiments against these three strains gave results that were clearly lower than 70% DNA-DNA similarity, and consequently confirmed that this new strain does not belong to a previously described species of the genus Comamonas. The major respiratory quinone was ubiquinone-8. The major fatty acids (>5%) were C16:0 (30.1%), summed feature 3 (C16:1ω6c and/or C16:1ω7c; 25.4%), summed feature 8 (C18:1ω6c and/or C18:1ω7c; 15.3%), C17:0 cyclo (7.4%) and C14:0 (5.8%). The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, unknown phospholipids and unknown lipids. Based on the phylogenetic analysis, DNA-DNA hybridization, whole-cell fatty acid composition as well as biochemical characteristics, strain YW1(T) was clearly distinguishable from all species of the genus Comamonas with validly published names and should be classified as a representative of a novel species of the genus Comamonas, for which the name Comamonas jiangduensis sp. nov. is proposed. The type strain is YW1(T) (=CCTCC AB 2012033(T)=KACC 16697(T)).


Assuntos
Comamonas/classificação , Filogenia , Microbiologia do Solo , Agricultura , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Comamonas/genética , Comamonas/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/análise , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Oryza/microbiologia , Fosfolipídeos/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Tensoativos/metabolismo , Ubiquinona
20.
Int J Syst Evol Microbiol ; 61(Pt 2): 255-258, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20207803

RESUMO

A bacterial strain, designated BF-3(T), was isolated from phenol-contaminated soil and investigated using a polyphasic taxonomic approach. Cells were Gram-reaction-negative, non-sporulating, non-motile, short rods. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BF-3(T) formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Comamonas; it showed highest sequence similarities to Comamonas aquatica LMG 2370(T) (96.8 %), C. nitrativorans DSM 13191(T) (96.4 %), C. odontotermitis LMG 23579(T) (96.4 %), C. kerstersii LMG 3475(T) (96.3 %), C. koreensis KCTC 12005(T) (96.1 %) and C. terrigena LMG 1253(T) (96.0 %). The major cellular fatty acids were C(16 : 0), C(18 : 1)/C(18 : 1)ω7c, C(17 : 0) cyclo and summed feature 3 (C(16 : 1)ω7c and/or iso-C(15 : 0) 2-OH). Based on the phylogenetic analysis, DNA-DNA hybridization, whole-cell fatty acid composition and biochemical characteristics, strain BF-3(T) was clearly distinct from type strains of other recognized species of the genus Comamonas and, as such, represents a novel species of the genus Comamonas, for which the name Comamonas zonglianii sp. nov. is proposed. The type strain is BF-3(T) (=CCTCC AB 209170(T) =DSM 22523(T)).


Assuntos
Comamonas/classificação , Fenol/química , Filogenia , Microbiologia do Solo , Poluentes do Solo/química , Técnicas de Tipagem Bacteriana , Composição de Bases , Comamonas/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/análise , Dados de Sequência Molecular , Fenótipo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Solo/química
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