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1.
Arch Microbiol ; 204(8): 525, 2022 Jul 27.
Artículo en Inglés | MEDLINE | ID: mdl-35895136

RESUMEN

Two novel Gram-stain-negative, aerobic, rod shaped bacterial strains BT290T and BT689T were isolated from soil collected in South Korea. Colony morphologies of both strains were circular and convex while the colors of BT290T and BT689T were light-pink and white, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that BT290T and BT689T belong to a distinct lineage within the genus Microvirga (family Methylobacteriaceae, order Rhizobiales, class Alphaproteobacteria, phylum Proteobacteria, kingdom Bacteria). The 16S rR NA gene sequence similarity between two strains was 97.9%. Both strains had the similar quinone system, with ubiquinone 10 (Q-10) as the major respiratory quinone. The major polar lipids of strains BT290T and BT689T were phosphatidylethanolamine (PE), diphosphatidylglycerol (DPG), phosphatidylcholine (PC), and phosphatidylglycerol (PG). The major cellular fatty acids of strain BT290T were C18:1 ω7c (58.2%) and C16:0 (17.7%), while those of strain BT689T were C18:1 ω7c (61.8%) and C16:0 (10.8%). On the bases of polyphasic analysis (phylogenetic, chemotaxonomic, and biochemical), strains BT290T and BT689T can be suggested as novel bacterial species within the genus Microvirga and the proposed names are Microvirga terrestris and Microvirga arvi, respectively. The type strain of Microvirga terrestris is BT290T (= KCTC 72367T = NBRC 114844T) and the type strain of Microvirga arvi is BT689T (= KACC 22016T = NBRC 114858T), respectively.


Asunto(s)
Alphaproteobacteria , Bradyrhizobiaceae , Methylobacteriaceae , Alphaproteobacteria/genética , Técnicas de Tipificación Bacteriana , Composición de Base , Bradyrhizobiaceae/genética , ADN Bacteriano/genética , Ácidos Grasos/análisis , Filogenia , Quinonas , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Suelo , Microbiología del Suelo
2.
Artículo en Inglés | MEDLINE | ID: mdl-35666677

RESUMEN

Two Gram-negative, rod-shaped, non-spore-forming bacteria, designated SM9T and SM2T, were isolated from Taklamakan Desert soil samples. Phylogenetic analysis based on the 16S rRNA gene sequences showed that strains SM9T and SM2T had the highest sequence similarity to the type strains Microvirga indica BCRC 80972T and Microvirga soli NBRC 112417T with similarity values of 98.2 and 97.7 %, respectively, and Microvirga was among the predominant genera in the desert soil. The draft genomes of these two strains were 4.56 Mbp (SM9T) and 5.08 Mbp (SM2T) long with 65.1 mol% (SM9T) and 63.5 mol% (SM2T) G+C content. To adapt to the desert environment, these two strains possessed pathways for the synthesis of stress metabolite trehalose. The major fatty acids (>5 %) included C18 : 1 ω9c in SM2T, but C16 : 0, C18 : 0 and C19 : 0 cyclo ω8c in SM9T, while the major menaquinone was ubiquinone 10 in both strains. The major polar lipids of SM9T and SM2T were phosphatidylglycerol, phosphatidylethanolamine and phospholipid. The average nucleotide identity and digital DNA-DNA hybridization results further indicated that strains SM9T and SM2T were distinguished from phylogenetically related species and represented two novel species within the genus Microvirga, for which the names Microvirga roseola sp. nov. (type strain SM2T=KCTC 72792T=CGMCC 1.17776T) and Microvirga lenta sp. nov. (type strain SM9T=KCTC 82729T=CCTCC AB 2021131T) are proposed.


Asunto(s)
Bradyrhizobiaceae , Suelo , Técnicas de Tipificación Bacteriana , Composición de Base , Bradyrhizobiaceae/genética , ADN Bacteriano/genética , Ácidos Grasos/química , Fosfolípidos/análisis , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Microbiología del Suelo
3.
Arch Microbiol ; 204(7): 390, 2022 Jun 14.
Artículo en Inglés | MEDLINE | ID: mdl-35699786

RESUMEN

Strain WGZ8T was isolated from a soil sample of Puerh tea garden in Pu'er city, Southwest China. The isolate was rod-shaped, Gram-stain negative, facultative anaerobic, non-motile. Growth occurred within 0-3.0% (w/v) NaCl (optimal concentration, 0-1.0%), pH 5.0-11.0 (optimal pH, 7.0) and 10-40 °C (optimal temperature, 28 °C). 16S rRNA gene sequence-based phylogenetic and phylogenomic analysis revealed that WGZ8T belonged to the genus Microvirga. Its major cellular fatty acids were C19:0 cyclo ω8c, C16:0, C18:1ω7c and/or C18:1ω6c. The profile of polar lipids included phosphatidyldimethylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol and phosphatidylglycerol. The only respiratory quinone was detected as ubiquinone 10 (Q-10). The genome size of strain WGZ8T was 5.17 MB, and the content of DNA G + C was 61 mol%. Based on the results of digital DNA-DNA hybridization and phenotypic results, strain WGZ8T could be concluded as a novel species of the genus Microvirga, for which the name Microvirga puerhi sp. nov. is proposed. The type strain is WGZ8T (= CGMCC 1.19171 T = JCM 35317 T).


Asunto(s)
Bradyrhizobiaceae , Methylobacteriaceae , Técnicas de Tipificación Bacteriana , Composición de Base , Bradyrhizobiaceae/genética , ADN Bacteriano/genética , Ácidos Grasos/análisis , Fosfolípidos/análisis , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Suelo , Microbiología del Suelo ,
4.
Arch Microbiol ; 204(7): 423, 2022 Jun 24.
Artículo en Inglés | MEDLINE | ID: mdl-35750895

RESUMEN

Two Gram-staining-negative, aerobic and rod-shaped strains, designated c23x22T and sex2T, were isolated from forest soil collected from Chebaling National Nature Reserve in Guangdong Province and Limu Mountain National Forest Park in Hainan Province, P. R. China, respectively. Phylogenetic analyses based on 16S rRNA gene sequences revealed that they belonged to the genus Microvirga, and strain c23 x22T was most closely related to 'Microvirga alba' KCTC 72385, while strain sex2T showed close relationship with Microvirga guangxiensis CGMCC 1.7666T. The average nucleotide identity and digital DNA-DNA hybridization values between strains c23 x22T and sex2T and their close relatives, 'M. alba' KCTC 72385 and M. guangxiensis CGMCC 1.7666T, were all below the threshold values for species delimitation. The predominant quinones of the two novel strains were ubiquinone 10, and the major fatty acids contained C19:0 cyclo ω8c and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). Their predominant polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. The phenotypic, genotypic and chemotaxonomic analyses clearly supported that strains c23 x 22T and sex2T represent two novel species of the genus Microvirga, for which the name Microvirga terricola sp. nov. (type strain c23 x 22T = GDMCC 1.1700T = KCTC 62432T) and Microvirga solisilvae sp. nov. (type strain sex2T = GDMCC 1.1651T = KACC 21311T) are proposed, respectively.


Asunto(s)
Bradyrhizobiaceae , Suelo , Técnicas de Tipificación Bacteriana , Composición de Base , Bradyrhizobiaceae/genética , ADN Bacteriano/genética , Ácidos Grasos/análisis , Bosques , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Microbiología del Suelo
5.
J Microbiol ; 60(2): 192-206, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-35102526

RESUMEN

Toxin-antitoxin (TA) systems are growth-controlling genetic elements consisting of an intracellular toxin protein and its cognate antitoxin. TA systems have been spread among microbial genomes through horizontal gene transfer and are now prevalent in most bacterial and archaeal genomes. Under normal growth conditions, antitoxins tightly counteract the activity of the toxins. Upon stresses, antitoxins are inactivated, releasing activated toxins, which induce growth arrest or cell death. In this study, among nine functional TA modules in Bosea sp. PAMC 26642 living in Arctic lichen, we investigated the functionality of BoHigBA2. BohigBA2 is located close to a genomic island and adjacent to flagellar gene clusters. The expression of BohigB2 induced the inhibition of E. coli growth at 37°C, which was more manifest at 18°C, and this growth defect was reversed when BohigA2 was co-expressed, suggesting that this BoHigBA2 module might be an active TA module in Bosea sp. PAMC 26642. Live/dead staining and viable count analyses revealed that the BoHigB2 toxin had a bactericidal effect, causing cell death. Furthermore, we demonstrated that BoHigB2 possessed mRNA-specific ribonuclease activity on various mRNAs and cleaved only mRNAs being translated, which might impede overall translation and consequently lead to cell death. Our study provides the insight to understand the cold adaptation of Bosea sp. PAMC 26642 living in the Arctic.


Asunto(s)
Antitoxinas/metabolismo , Toxinas Bacterianas/metabolismo , Bradyrhizobiaceae/genética , Escherichia coli/crecimiento & desarrollo , Escherichia coli/metabolismo , Sistemas Toxina-Antitoxina , Antitoxinas/genética , Regiones Árticas , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/genética , Escherichia coli/genética , Regulación Bacteriana de la Expresión Génica , Islas Genómicas , Familia de Multigenes , ARN Mensajero/metabolismo
6.
RNA ; 27(11): 1374-1389, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34429367

RESUMEN

Toxin-antitoxin (TA) systems are genetic modules composed of a toxin interfering with cellular processes and its cognate antitoxin, which counteracts the activity of the toxin. TA modules are widespread in bacterial and archaeal genomes. It has been suggested that TA modules participate in the adaptation of prokaryotes to unfavorable conditions. The Bosea sp. PAMC 26642 used in this study was isolated from the Arctic lichen Stereocaulon sp. There are 12 putative type II TA loci in the genome of Bosea sp. PAMC 26642. Of these, nine functional TA systems have been shown to be toxic in Escherichia coli The toxin inhibits growth, but this inhibition is reversed when the cognate antitoxin genes are coexpressed, indicating that these putative TA loci were bona fide TA modules. Only the BoVapC1 (AXW83_01405) toxin, a homolog of VapC, showed growth inhibition specific to low temperatures, which was recovered by the coexpression of BoVapB1 (AXW83_01400). Microscopic observation and growth monitoring revealed that the BoVapC1 toxin had bacteriostatic effects on the growth of E. coli and induced morphological changes. Quantitative real time polymerase chain reaction and northern blotting analyses showed that the BoVapC1 toxin had a ribonuclease activity on the initiator tRNAfMet, implying that degradation of tRNAfMet might trigger growth arrest in E. coli Furthermore, the BoVapBC1 system was found to contribute to survival against prolonged exposure at 4°C. This is the first study to identify the function of TA systems in cold adaptation.


Asunto(s)
Antitoxinas/metabolismo , Toxinas Bacterianas/metabolismo , Bradyrhizobiaceae/crecimiento & desarrollo , Escherichia coli/crecimiento & desarrollo , Regulación Bacteriana de la Expresión Génica , ARN de Transferencia de Metionina/metabolismo , Sistemas Toxina-Antitoxina/genética , Antitoxinas/genética , Proteínas Bacterianas , Toxinas Bacterianas/genética , Bradyrhizobiaceae/genética , Bradyrhizobiaceae/aislamiento & purificación , Bradyrhizobiaceae/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Líquenes/fisiología , Operón , Regiones Promotoras Genéticas
7.
Molecules ; 26(10)2021 May 12.
Artículo en Inglés | MEDLINE | ID: mdl-34066118

RESUMEN

Lignin, which is a component of wood, is difficult to degrade in nature. However, serious decay caused by microbial consortia can happen to wooden antiques during the preservation process. This study successfully screened four microbial consortia with lignin degradation capabilities (J-1, J-6, J-8 and J-15) from decayed wooden antiques. Their compositions were identified by genomic sequencing, while the degradation products were analyzed by GC-MS. The lignin degradation efficiency of J-6 reached 54% after 48 h with an initial lignin concentration of 0.5 g/L at pH 4 and rotation speed of 200 rpm. The fungal consortium of J-6 contained Saccharomycetales (98.92%) and Ascomycota (0.56%), which accounted for 31% of the total biomass. The main bacteria in J-6 were Shinella sp. (47.38%), Cupriavidus sp. (29.84%), and Bosea sp. (7.96%). The strongest degradation performance of J-6 corresponded to its composition, where Saccharomycetales likely adapted to the system and improved lignin degradation enzymes activities, and the abundant bacterial consortium accelerated lignin decomposition. Our work demonstrated the potential utilization of microbial consortia via the synergy of microbial consortia, which may overcome the shortcomings of traditional lignin biodegradation when using a single strain, and the potential use of J-6 for lignin degradation/removal applications.


Asunto(s)
Lignina/metabolismo , Consorcios Microbianos/genética , Madera/metabolismo , Madera/microbiología , Ascomicetos/genética , Biodegradación Ambiental , Biomasa , Bradyrhizobiaceae/genética , Cupriavidus/genética , ADN Bacteriano/genética , ADN de Hongos/genética , Cromatografía de Gases y Espectrometría de Masas , Lacasa/metabolismo , Peroxidasas/metabolismo , Rhizobiaceae/genética , Saccharomycetales/genética , Análisis de Secuencia de ADN
8.
Int J Mol Sci ; 21(21)2020 Oct 27.
Artículo en Inglés | MEDLINE | ID: mdl-33121154

RESUMEN

The free-living Gram-negative bacterium Oligotropha carboxidovorans (formerly: Pseudomonas carboxydovorans), isolated from wastewater, is able to live in aerobic and, facultatively, in autotrophic conditions, utilizing carbon monoxide or hydrogen as a source of energy. The structure of O. carboxidovorans lipid A, a hydrophobic part of lipopolysaccharide, was studied using NMR spectroscopy and high-resolution mass spectrometry (MALDI-ToF MS) techniques. It was demonstrated that the lipid A backbone is composed of two d-GlcpN3N residues connected by a ß-(1→6) glycosidic linkage, substituted by galacturonic acids (d-GalpA) at C-1 and C-4' positions. Both diaminosugars are symmetrically substituted by 3-hydroxy fatty acids (12:0(3-OH) and 18:0(3-OH)). Ester-linked secondary acyl residues (i.e., 18:0, and 26:0(25-OH) and a small amount of 28:0(27-OH)) are located in the distal part of lipid A. These very long-chain hydroxylated fatty acids (VLCFAs) were found to be almost totally esterified at the (ω-1)-OH position with malic acid. Similarities between the lipid A of O. carboxidovorans and Mesorhizobium loti, Rhizobium leguminosarum, Caulobacter crescentus as well as Aquifex pyrophylus were observed and discussed from the perspective of the genomic context of these bacteria.


Asunto(s)
Bradyrhizobiaceae/metabolismo , Ácidos Hexurónicos/química , Lípido A/química , Malatos/química , Sustitución de Aminoácidos , Bradyrhizobiaceae/química , Bradyrhizobiaceae/genética , Secuencia de Carbohidratos , Lípido A/genética , Lípido A/metabolismo , Espectroscopía de Resonancia Magnética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
9.
Arch Microbiol ; 202(10): 2727-2738, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32734321

RESUMEN

Tuber species may be regarded as complex microhabitats hosting diverse microorganisms inside their fruiting bodies. Here, we investigated the structure of microbial communities inhabiting the gleba of wild growing (in stands) T. aestivum, using Illumina sequencing and culture-based methods. The two methods used in combination allowed to extract more information on complex microbiota of Tuber aestivum gleba. Analysis of the V3-V4 region of 16S rDNA identified nine phyla of bacteria present in the gleba of T. aestivum ascomata, mostly Proteobacteria from the family Bradyrhizobiaceae. Our results ideally match the earlier data for other Tuber species where the family Bradyrhizobiaceae was the most represented. The ITS1 region of fungal rDNA represented six alien fungal species belonging to three phyla. To complement the metagenomic analysis, cultivable fungi and bacteria were obtained from the gleba of the same T. aestivum fruiting bodies. The identified fungi mostly belong to the phylum Basidiomycota and same to Ascomycota. Analysis of cultivable bacteria revealed that all the specimens were colonized by different strains of Bacillus. Fungal community inhabiting T. aestivum fruiting bodies was never shown before.


Asunto(s)
Ascomicetos/fisiología , Bacillus/aislamiento & purificación , Basidiomycota/aislamiento & purificación , Bradyrhizobiaceae/aislamiento & purificación , Cuerpos Fructíferos de los Hongos/fisiología , Bacillus/clasificación , Bacillus/genética , Basidiomycota/clasificación , Basidiomycota/genética , Bradyrhizobiaceae/clasificación , Bradyrhizobiaceae/genética , ADN Ribosómico/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Microbiota
10.
Mol Plant Microbe Interact ; 33(10): 1232-1241, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32686981

RESUMEN

A collection of rhizobial strains isolated from root nodules of the narrowly endemic legume species Oxytropis erecta, O. anadyrensis, O. kamtschatica, and O. pumilio originating from the Kamchatka Peninsula (Russian Federation) was obtained. Analysis of the 16S ribosomal RNA gene sequence showed a significant diversity of isolates belonging to families Rhizobiaceae (genus Rhizobium), Phyllobacteriaceae (genera Mesorhizobium, Phyllobacterium), and Bradyrhizobiaceae (genera Bosea, Tardiphaga). A plant nodulation assay showed that only strains belonging to genus Mesorhizobium could form nitrogen-fixing nodules on Oxytropis plants. The strains M. loti 582 and M. huakuii 583, in addition to symbiotic clusters, possessed genes of the type III and type VI secretion systems (T3SS and T6SS, respectively), which can influence the host specificity of strains. These strains formed nodules of two types (elongated and rounded) on O. kamtschatica roots. We suggest this phenomenon may result from Nod factor-dependent and -independent nodulation strategies. The obtained strains are of interest for further study of the T3SS and T6SS gene function and their role in the development of rhizobium-legume symbiosis. The prospects of using rhizobia having both gene systems related to symbiotic and nonsymbiotic nodulation strategies to enhance the efficiency of plant-microbe interactions by expanding the host specificity and increasing nodulation efficiency are discussed.


Asunto(s)
Bradyrhizobiaceae , Mesorhizobium , Oxytropis/microbiología , Rhizobium , Simbiosis , Sistemas de Secreción Tipo III/genética , Sistemas de Secreción Tipo VI/genética , Bradyrhizobiaceae/genética , Mesorhizobium/genética , Filogenia , ARN Ribosómico 16S/genética , Rhizobium/genética , Nódulos de las Raíces de las Plantas/microbiología
11.
ACS Synth Biol ; 9(6): 1426-1440, 2020 06 19.
Artículo en Inglés | MEDLINE | ID: mdl-32379961

RESUMEN

Due to climate change and worldwide pollution, development of highly sustainable routes for industrial production of basic and specialty chemicals is critical nowadays. One possible approach is the use of CO2- and CO-utilizing microorganisms in biotechnological processes to produce value-added compounds from synthesis gas (mixtures of CO2, CO, and H2) or from C1-containing industrial waste gases. Such syngas fermentation processes have already been established, e.g., biofuel production using strictly anaerobic acetogenic bacteria. However, aerobic processes may be favorable for the formation of more costly (ATP-intensive) products. Oligotropha carboxidovorans strain OM5 is an aerobic carboxidotrophic bacterium and potentially a promising candidate for such processes. We here performed RNA-Seq analysis comparing cells of this organism grown heterotrophically with acetate or autotrophically with CO2, CO, and H2 as carbon and energy source and found a variety of chromosomally and of native plasmid-encoded genes to be highly differentially expressed. In particular, genes and gene clusters encoding proteins required for autotrophic growth (CO2 fixation via Calvin-Benson-Bassham cycle), for CO metabolism (CO dehydrogenase), and for H2 utilization (hydrogenase), all located on megaplasmid pHCG3, were much higher expressed during autotrophic growth with synthesis gas. Furthermore, we successfully established reproducible transformation of O. carboxidovoransvia electroporation and developed gene deletion and gene exchange protocols via two-step recombination, enabling inducible and stable expression of heterologous genes as well as construction of defined mutants of this organism. Thus, this study marks an important step toward metabolic engineering of O. carboxidovorans and effective utilization of C1-containing gases with this organism.


Asunto(s)
Bradyrhizobiaceae/genética , Gases/metabolismo , Genes Bacterianos , Ingeniería Genética/métodos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Dióxido de Carbono/metabolismo , Monóxido de Carbono/metabolismo , Edición Génica , Hidrógeno/metabolismo , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Familia de Multigenes , Oxidorreductasas/genética , Oxidorreductasas/metabolismo
12.
J Microbiol ; 58(7): 606-613, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32323197

RESUMEN

Crystal structures of enoyl-coenzyme A (CoA) isomerase from Bosea sp. PAMC 26642 (BoECI) and enoyl-CoA hydratase from Hymenobacter sp. PAMC 26628 (HyECH) were determined at 2.35 and 2.70 Å resolution, respectively. BoECI and HyECH are members of the crotonase superfamily and are enzymes known to be involved in fatty acid degradation. Structurally, these enzymes are highly similar except for the orientation of their C-terminal helix domain. Analytical ultracentrifugation was performed to determine the oligomerization states of BoECI and HyECH revealing they exist as trimers in solution. However, their putative ligand-binding sites and active site residue compositions are dissimilar. Comparative sequence and structural analysis revealed that the active site of BoECI had one glutamate residue (Glu135), this site is occupied by an aspartate in some ECIs, and the active sites of HyECH had two highly conserved glutamate residues (Glu118 and Glu138). Moreover, HyECH possesses a salt bridge interaction between Glu98 and Arg152 near the active site. This interaction may allow the catalytic Glu118 residue to have a specific conformation for the ECH enzyme reaction. This salt bridge interaction is highly conserved in known bacterial ECH structures and ECI enzymes do not have this type of interaction. Collectively, our comparative sequential and structural studies have provided useful information to distinguish and classify two similar bacterial crotonase superfamily enzymes.


Asunto(s)
Bacteroidetes/enzimología , Bradyrhizobiaceae/enzimología , Dodecenoil-CoA Isomerasa/metabolismo , Enoil-CoA Hidratasa/metabolismo , Secuencia de Aminoácidos , Bacteroidetes/genética , Sitios de Unión/genética , Bradyrhizobiaceae/genética , Dominio Catalítico/genética , Cristalografía por Rayos X , Ácidos Grasos/metabolismo , Modelos Moleculares , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Ultracentrifugación
13.
Appl Environ Microbiol ; 85(24)2019 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-31604771

RESUMEN

Many Gram-negative bacteria employ N-acylhomoserine lactones (AHLs) as quorum-sensing (QS) signal molecules to regulate virulence expression in a density-dependent manner. Quorum quenching (QQ) via enzymatic inactivation of AHLs is a promising strategy to reduce bacterial infections and drug resistance. Herein, a thermostable AHL lactonase (AidB), which could degrade different AHLs, with or without a substitution of carbonyl or hydroxyl at the C-3 position, was identified from the soil bacterium Bosea sp. strain F3-2. Ultrahigh-performance liquid chromatography analysis demonstrated that AidB is an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone (HSL) ring. AidB was thermostable in the range 30 to 80°C and showed maximum activity after preincubation at 60°C for 30 min. The optimum temperature of AidB was 60°C, and the enzyme could be stably stored in double-distilled water (ddH2O) at 4°C or room temperature. AidB homologs were found only in Rhizobiales and Rhodospirillales of the Alphaproteobacteria AidB from Agrobacterium tumefaciens and AidB from Rhizobium multihospitium (with amino acid identities of 50.6% and 52.8% to AidB, respectively) also showed thermostable AHL degradation activity. When introduced into bacteria, plasmid-expressed AidB attenuated pyocyanin production by Pseudomonas aeruginosa PAO1 and the pathogenicity of Pectobacterium carotovorum subsp. carotovorum Z3-3, suggesting that AidB is a potential therapeutic agent by degrading AHLs.IMPORTANCE A quorum-sensing system using AHLs as the signal in many bacterial pathogens is a critical virulence regulator and an attractive target for anti-infective drugs. In this work, we identified a novel AHL lactonase, AidB, from a soil bacterial strain, Bosea sp. F3-2. The expression of aidB reduced the production of AHL signals and QS-dependent virulence factors in Pseudomonas aeruginosa and Pectobacterium carotovorum The homologs of AidB with AHL-degrading activities were found only in several genera belonging to the Alphaproteobacteria Remarkably, AidB is a thermostable enzyme that retained its catalytic activity after treatment at 80°C for 30 min and exhibits reliable storage stability at both 4°C and room temperature. These properties might make it more suitable for practical application.


Asunto(s)
Bradyrhizobiaceae/enzimología , Bradyrhizobiaceae/metabolismo , Hidrolasas de Éster Carboxílico/metabolismo , 4-Butirolactona/análogos & derivados , Acil-Butirolactonas/metabolismo , Agrobacterium tumefaciens/metabolismo , Secuencia de Aminoácidos , Bacterias/metabolismo , Proteínas Bacterianas , Bradyrhizobiaceae/genética , Estabilidad de Enzimas , Pectobacterium carotovorum/metabolismo , Pseudomonas aeruginosa/metabolismo , Piocianina/metabolismo , Percepción de Quorum , Virulencia , Factores de Virulencia/metabolismo , Secuenciación Completa del Genoma
14.
J Hazard Mater ; 359: 527-534, 2018 10 05.
Artículo en Inglés | MEDLINE | ID: mdl-30086523

RESUMEN

Arsenic (As) and antinomy (Sb) usually coexist in natural environments where both of them pollute soils and water. Microorganisms that oxidize arsenite [As(III)] and tolerate Sb have great potential in As and Sb bioremediation, In this study, a Gram-negative bacterial strain, Bosea sp. AS-1, was isolated from a mine slag sample collected in Xikuangshan Sb mine in China. AS-1 could tolerate 120 mM of As(III) and 50 mM of antimonite [Sb(III)]. It could also oxidize 2 mM of As(III) or Sb(III) completely under heterotrophic and aerobic conditions. Interestingly, strain AS-1 preferred to oxidize As(III) with yeast extract as the carbon source, whereas Sb(III) oxidation was favored with lactate in the medium. Genomic analysis of AS-1 confirmed the presence of several gene islands for As resistance and oxidation. Notably, a system of AS-1 and goethite was found to be able to remove 99% of the As with the initial concentration of 500 µg/L As(III) and 500 µg/L Sb(III), which suggests the potential of this approach for As removal in environments especially with the presence of high Sb.


Asunto(s)
Antimonio/metabolismo , Arsenitos/metabolismo , Bradyrhizobiaceae/metabolismo , Contaminantes Ambientales/metabolismo , Antimonio/química , Arsénico/química , Arsénico/metabolismo , Arsenitos/química , Biodegradación Ambiental , Bradyrhizobiaceae/genética , Contaminantes Ambientales/química , Compuestos de Hierro/química , Minerales/química , Oxidación-Reducción , ARN Ribosómico 16S/genética
15.
Syst Appl Microbiol ; 41(6): 641-649, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30145046

RESUMEN

Given that phosphate supplies may diminish and become uneconomic to mine after 2020, there is a compelling need to develop alternative industries to support the population on Christmas Island. Former mine sites could be turned into productive agricultural land, however, large-scale commercial agriculture has never been attempted, and, given the uniqueness of the island, the diversity of rhizobia prior to introducing legumes needed evaluation. Therefore, 84 rhizobia isolates were obtained from nine different hosts, both crop and introduced legumes, located at seven sites across the island. Based on 16S rRNA and recA gene sequence analysis, the isolates grouped into 13 clades clustering within the genus Bradyrhizobium, Ensifer, Cupriavidus and Rhizobium. According to the sequences of their symbiosis genes nodC and nifH, the isolates were classified into 12 and 11 clades, respectively, and clustered closest to tropical or crop legume isolates. Moreover, the symbiosis gene phylogeny and Multi Locus Sequence Analysis gene phylogeny suggested vertical transmission in the Alpha-rhizobia but horizontal transmission within the Beta-rhizobia. Furthermore, this study provides evidence of a large diversity of endemic rhizobia associated with both crop and introduced legumes, and highlights the necessity of inoculation for common bean, chickpea and soybean on the Island.


Asunto(s)
Bradyrhizobiaceae/clasificación , Fabaceae/microbiología , Minería , Rhizobiaceae/clasificación , Nódulos de las Raíces de las Plantas/microbiología , Agricultura , Australia , Bradyrhizobiaceae/genética , Bradyrhizobiaceae/aislamiento & purificación , ADN Bacteriano/genética , Genes Bacterianos , Fosfatos , Filogenia , ARN Ribosómico 16S/genética , Rhizobiaceae/genética , Rhizobiaceae/aislamiento & purificación , Simbiosis
16.
Curr Issues Mol Biol ; 24: 71-102, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28686569

RESUMEN

Spatial and temporal variations related to hydric seasonality in abundance and diversity of denitrifier communities were examined in sediments taken from two sites differing in nitrate concentration along a stream Doñana National Park during a 3-year study. We found a positive relationship between the relative abundance of denitrifiers, determined as narG, napA, nirK, nirS and nosZ denitrification genes, and sediment nitrate content, with similar spatial and seasonal variations. However, we did not find association between denitrification activity and the community structure of denitrifiers. Because nosZ showed the strongest correlation with the content of nitrate in sediments, we used this gene as a molecular marker to construct eight genomic libraries. Analysis of these genomic libraries revealed that diversity of the nosZ-bearing communities was higher in the site with higher nitrate content. Regardless of nitrate concentration in the sediments, the Bradyrhizobiaceae and Rhodocyclaceae were the most abundant families. On the contrary, Rhizobiaceae was exclusively present in sediments with higher nitrate content. Results showed that differences in sediment nitrate concentration affect the composition and diversityof nosZ-bearing communities.


Asunto(s)
Desnitrificación , Sedimentos Geológicos/microbiología , Nitratos/metabolismo , Microbiología del Suelo , Proteínas Bacterianas/genética , Biodiversidad , Bradyrhizobiaceae/genética , Bradyrhizobiaceae/aislamiento & purificación , Bradyrhizobiaceae/metabolismo , Sedimentos Geológicos/análisis , Nitratos/análisis , Nitrito Reductasas/genética , Filogenia , Rhodocyclaceae/genética , Rhodocyclaceae/aislamiento & purificación , Rhodocyclaceae/metabolismo , Análisis Espacio-Temporal
17.
Int J Syst Evol Microbiol ; 67(1): 127-132, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27902203

RESUMEN

An aerobic, Gram-stain-negative, catalase-positive and oxidase-negative, non-motile, non-spore-forming, rod-shaped, pink-pigmented bacterium designated strain R491T was isolated from soil. Flexirubin-type pigments were absent. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain R491T formed a lineage within the family Methylobacteriaceae of the phylum Proteobacteria that was distinct from various species of the genus Microvirga, including Microvirgaaerilata 5420S-16T (97.83 % 16S rRNA gene sequence similarity), Microvirga zambiensis WSM3693T (97.76 %), Microvirga flocculans ATCC BAA-817T (97.41 %), and Microvirga lupini Lut6T, Microvirga subterranea DSM 14364T, Microvirga vignae BR3299T, and Microvirga guangxiensis 25BT (96.99 %). The major polar lipids of strain R491T were phosphatidylcholine, phosphatidylglycerol and phosphatidylethanolamine. The major cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c; 71.2 %), C16 : 0 (12.0 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c; 4.7 %), and C18 : 0 (4.2 %). The DNA G+C content of strain R491T was 61.8 mol%. DNA-DNA hybridization values between strain R491T and other members of the genus Microvirga ranged from 27 to 57 %. On the basis of phenotypic, genotypic and phylogenetic analyses, strain R491T represents a novel species of the genus Microvirga, for which the name Microvirga soli sp. nov. is proposed. The type strain is R491T (=KEMB 9005-408T=KACC 18969T=NBRC 112417T).


Asunto(s)
Methylobacteriaceae/clasificación , Filogenia , Microbiología del Suelo , Técnicas de Tipificación Bacteriana , Composición de Base , Bradyrhizobiaceae/genética , ADN Bacteriano/genética , Ácidos Grasos/análisis , Methylobacteriaceae/genética , Methylobacteriaceae/aislamiento & purificación , Hibridación de Ácido Nucleico , Pigmentación , ARN Ribosómico 16S/genética , República de Corea , Análisis de Secuencia de ADN
18.
Can J Microbiol ; 61(6): 399-408, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25941999

RESUMEN

The adsorption behavior of Cr(VI) ions from aqueous solution by a chromium-tolerant strain was studied through batch experiments. An isolate designated Zer-1 was identified as a species of Bosea on the basis of 16S rRNA results. It showed a maximum resistance to 550 mg·L(-1) Cr(VI). The effects of 3 important operating parameters, initial solution pH, initial Cr(VI) concentration, and biomass dose, were investigated by central composite design. On the basis of response surface methodology results, maximal removal efficiency of Cr(VI) was achieved under the following conditions: pH, 2.0; initial concentration of metal ions, 55 mg·L(-1); and biomass dose, 2.0 g·L(-1). Under the optimal conditions, the maximum removal efficiency of Cr(VI) ions was found to be nearly 98%. The experimental data exhibited a better fit with the Langmuir model than the Freundlich model. The biosorption mechanisms were investigated with pseudo-first-order, pseudo-second-order, and intraparticle diffusion kinetics models. These results revealed that biosorption of Cr(VI) onto bacterial biomass could be an alternative method for the removal of metal ions from aqueous solution.


Asunto(s)
Bradyrhizobiaceae/metabolismo , Cromo/metabolismo , Restauración y Remediación Ambiental/instrumentación , Contaminantes Químicos del Agua/metabolismo , Adsorción , Biodegradación Ambiental , Bradyrhizobiaceae/clasificación , Bradyrhizobiaceae/genética , Bradyrhizobiaceae/aislamiento & purificación , Cromo/química , Concentración de Iones de Hidrógeno , Iones/metabolismo , Cinética , Datos de Secuencia Molecular , Filogenia , Suelo/química , Microbiología del Suelo , Contaminantes Químicos del Agua/química
19.
Arch Microbiol ; 197(7): 889-98, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26013968

RESUMEN

Eleven extra-slow-growing strains were isolated from nodules of the relict legume Vavilovia formosa growing in North Ossetia (Caucasus) and Armenia. All isolates formed a single rrs cluster together with the type strain Tardiphaga robiniae LMG 26467(T), while the sequencing of the 16S-23S rDNA intergenic region (ITS) and housekeeping genes glnII, atpD, dnaK, gyrB, recA and rpoB divided them into three groups. North Ossetian isolates (in contrast to the Armenian ones) were clustered separately from the type strain LMG 26467(T). However, all isolates were classified as T. robiniae because the DNA-DNA relatedness between them and the type strain LMG 26467(T) was 69.6% minimum. Two symbiosis-related genes (nodM and nodT) were amplified in all isolated Tardiphaga strains. It was shown that the nodM gene phylogeny is similar to that of ITS and housekeeping genes. The presence of the other symbiosis-related genes in described Tardiphaga strains, which is recently described genus of rhizobia, as well as their ability to form nodules on any plants are under investigation.


Asunto(s)
Bradyrhizobiaceae/clasificación , Bradyrhizobiaceae/fisiología , Fabaceae/microbiología , Técnicas de Tipificación Bacteriana , Bradyrhizobiaceae/genética , Bradyrhizobiaceae/crecimiento & desarrollo , Bradyrhizobiaceae/aislamiento & purificación , ADN Bacteriano/genética , Filogenia , ARN Ribosómico 16S/genética , Simbiosis/genética , Taiwán
20.
PLoS One ; 10(4): e0121274, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25874801

RESUMEN

An old world fruit bat Pteropus giganteus, held in captivity and suffering from necrosis of its wing digits, failed to respond to antibiotic therapy and succumbed to the infection. Samples submitted to the National Centre for Foreign Animal Disease were tested for viral infection. Vero E6 cells exhibited minor but unique cytopathic effects on second blind passage, and full CPE by passage four. Utilizing an unbiased random amplification technique from cell culture supernatant, we identified a bacterium belonging to the Bradyrhizobiaceae. Purification of cell culture supernatant on TY media revealed a slow growing bacterial isolate. In this study using electron microscopy, 16S rRNA gene analysis and whole genome sequencing, we identify a novel bacterial species associated with the site of infection belonging to the genus Afipia. This genus of bacteria is very diverse, with only a limited number of species characterized. Afipia felis, previously described as the etiological agent to cause cat scratch disease, and Afipia septicemium, most recently shown to cause disease in humans, highlight the potential for members of this genus to form a branch of opportunistic pathogens within the Bradyrhizobiaceae. Increased utilization of next generation sequencing and genomics will aid in classifying additional members of this intriguing bacterial genera.


Asunto(s)
Bradyrhizobiaceae/aislamiento & purificación , Quirópteros/microbiología , Necrosis/microbiología , Alas de Animales/microbiología , Afipia/patogenicidad , Animales , Bradyrhizobiaceae/genética , Bradyrhizobiaceae/patogenicidad , Enfermedad por Rasguño de Gato/genética , Enfermedad por Rasguño de Gato/microbiología , Gatos , Genoma Bacteriano , Humanos , Necrosis/fisiopatología , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Alas de Animales/fisiopatología
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