RESUMEN
High-salt conditions reduce the efficiency of electricity generation and nitrogen removal in microbial fuel cells (MFCs). In this work, we propose a three-phase single-chamber MFC (TP-MFC) by setting up a phase with immobilized cells in a conventional bipolar single-chamber MFC (common MFC). Cells from Halomonas were used as the immobilized phase, because these cells secrete the compatible solute ectoine and exhibit simultaneous nitrification and denitrification (SND). This enhanced the efficiency of SND and subsequent electricity generation under high-salt conditions. The average voltage of TP-MFC generated during the stable period in the presence of 30 g/L NaCl was 439.3 mV, which was 55.2% higher than that generated in common MFC. In addition, the N-removal rate of TP-MFC at 72 h was 63.4%, which was 38.4% higher than that of common MFC. The 16S rRNA diversity analysis showed an improved abundance of Pseudomonas, Acinetobacter, Alcaligenes, and Halomonas in TP-MFC, indicating that the ectoine secreted by immobilized Halomonas conferred substantial salt-tolerance on the electrogenic bacteria growing in a high-salt environment. This paper establishes an efficient and convenient method for improving the salt tolerance of microbial flora in MFCs, which is of great significance for the application of MFCs in high-strength wastewater treatment.
Asunto(s)
Fuentes de Energía Bioeléctrica/microbiología , Halomonas/metabolismo , Células Inmovilizadas/citología , Células Inmovilizadas/metabolismo , Desnitrificación , Electricidad , Diseño de Equipo , Halomonas/citología , Sales (Química)/metabolismoRESUMEN
Two moderately halophilic strains SBS 10T and SSO 06 were isolated from the saltern crystallizer ponds of the hypersaline Sambhar Salt Lake in India. Strains were aerobic, Gram-stain-negative, and rod shaped. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that two strains belong to the genus Halomonas in the Gammaproteobacteria, with highest 16S rRNA gene sequence similarities with Halomonas gudaonensis LMG 23610T (98.2% similarity) and Halomonas campaniensis 5AGT (99.0% similarity). Strains grew optimally at 37 °C, pH 7.5-8.0 in the presence of 5-8% (w/v) NaCl. The major fatty acids of the strain SBS 10T were C18:1ω7c (54.37%), C16:0 (25.69%), C16:1 × 7c/C16:1 × 6c (13.28%), and C12:0 (1.21%). The G+C content was 63.6 mol % (Tm). Phenotypic features, fatty acids profile, and DNA G+C content supported placement of the strain SBS 10T in the genus Halomonas having distinct characteristics with related strains. Analysis of the housekeeping genes: gryB and rpoD and in silico DNA-DNA hybridization between the strain SBS 10T and its type strain Halomonas gudaonensis (LMG 23610T) further revealed the strain SBS 10T to be a distinct species. On the basis of the phenotypic, chemotaxonomic and phylogenetic analysis, the strain SBS 10T is considered to represent a novel species for which the name Halomonas sambharensis is proposed. The type strain is SBS 10T (= MTCC 12313T = LMG 30344T).
Asunto(s)
Halomonas/clasificación , Halomonas/fisiología , Estanques/microbiología , Sales (Química)/metabolismo , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos , Genes Esenciales/genética , Halomonas/química , Halomonas/citología , Concentración de Iones de Hidrógeno , India , Lagos , Hibridación de Ácido Nucleico , Fenotipo , Filogenia , Estanques/química , ARN Ribosómico 16S/genética , Sales (Química)/análisis , Análisis de Secuencia de ADN , Especificidad de la Especie , TemperaturaRESUMEN
Two Gram-negative strains obtained from tank water in a scallop hatchery in Norway, were phenotypically and genotypically characterized in order to clarify their taxonomic position. On the basis of 16S rRNA gene sequence analysis, these isolates, ATF 5.2T and ATF 5.4T, were included in the genus Halomonas, being their closest relatives H. smyrnensis and H. taeanensis, with similarities of 98.9% and 97.7%, respectively. Sequence analysis of the housekeeping genes atpA, ftsZ, gyrA, gyrB, mreB, rpoB, rpoD, rpoE, rpoH, rpoN and rpoS clearly differentiated the isolates from the currently described Halomonas species, and the phylogenetic analysis using concatenated sequences of these genes located them in two robust and independent branches. DNA-DNA hybridization (eDDH) percentage, together with average nucleotide identity (ANI), were calculated using the complete genome sequences of the strains, and demonstrate that the isolates constitute two new species of Halomonas, for which the names of Halomonas borealis sp. nov. and Halomonas niordiana sp. nov. are proposed, with type strains ATF 5.2T (=CECT 9780T=LMG 31367T) and ATF 5.4T (=CECT 9779T=LMG 31227T), respectively.
Asunto(s)
Halomonas/clasificación , Agua de Mar/microbiología , Proteínas Bacterianas/genética , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/análisis , Genes Esenciales/genética , Genoma Bacteriano/genética , Halomonas/química , Halomonas/citología , Halomonas/fisiología , Noruega , Hibridación de Ácido Nucleico , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Especificidad de la Especie , Ubiquinona/químicaRESUMEN
Polyhydroxyalkanoates (PHA) are microbial polyesters which accumulate as intracellular granules in numerous prokaryotes and mainly serve as storage materials; beyond this primary function, PHA also enhance the robustness of bacteria against various stress factors. We have observed that the presence of PHA in bacterial cells substantially enhances their ability to maintain cell integrity when suddenly exposed to osmotic imbalances. In the case of the non-halophilic bacterium Cupriavidus necator, the presence of PHA decreased plasmolysis-induced cytoplasmic membrane damage during osmotic up-shock, which subsequently enabled the cells to withstand subsequent osmotic downshock. In contrast, sudden induction of osmotic up- and subsequent down-shock resulted in massive hypotonic lysis of non-PHA containing cells as determined by Transmission Electron Microscopy and Thermogravimetrical Analysis. Furthermore, a protective effect of PHA against hypotonic lysis was also observed in the case of the halophilic bacterium Halomonas halophila; here, challenged PHA-rich cells were capable of retaining cell integrity more effectively than their PHA-poor counterparts. Hence, it appears that the fact that PHA granules, as an added value to their primary storage function, protect halophiles from the harmful effect of osmotic down-shock might explain why PHA accumulation is such a common feature among halophilic prokaryotes. The results of this study, apart from their fundamental importance, are also of practical biotechnological significance: because PHA-rich bacterial cells are resistant to osmotic imbalances, they could be utilized in in-situ bioremediation technologies or during enrichment of mixed microbial consortia in PHA producers under conditions of fluctuating salinity.
Asunto(s)
Bacterias/citología , Bacterias/metabolismo , Cupriavidus necator/citología , Halomonas/citología , Ósmosis , Polihidroxialcanoatos/farmacología , Bacterias/efectos de los fármacos , Cupriavidus necator/efectos de los fármacos , Cupriavidus necator/metabolismo , Cupriavidus necator/ultraestructura , Halomonas/efectos de los fármacos , Halomonas/metabolismo , Halomonas/ultraestructura , Viabilidad Microbiana/efectos de los fármacos , Temperatura , TermogravimetríaRESUMEN
Traditional microbial chassis, including Escherichia coli, Bacillus subtilis, Ralstonia eutropha, and Pseudomonas putida, are grown under neutral pH and mild osmotic pressure for production of chemicals and materials. They tend to be contaminated easily by many microorganisms. To address this issue, next-generation industrial biotechnology employing halophilic Halomonas spp. has been developed for production of bioplastics polyhydroxyalkanoates (PHAs) and other chemicals. Halomonas spp. that can be grown contamination free under open and unsterile condition at alkali pH and high NaCl have been engineered to produce several PHA polymers in elongated or enlarged cells. New pathways can also be constructed both in plasmids and on chromosomes for Halomonas spp. Synthetic biology approaches and parts have been developed for Halomonas spp., allowing better control of their growth and product formation as well as morphology adjustment. Halomonas spp. and their synthetic biology will play an increasingly important role for industrial production of large volume chemicals.
Asunto(s)
Halomonas/genética , Microbiología Industrial/métodos , Ingeniería Metabólica/métodos , Polihidroxialcanoatos/genética , Vías Biosintéticas , Cromosomas Bacterianos/genética , Cromosomas Bacterianos/metabolismo , Fermentación , Halomonas/citología , Halomonas/crecimiento & desarrollo , Halomonas/metabolismo , Polihidroxialcanoatos/metabolismo , Biología Sintética/métodosRESUMEN
BACKGROUND: Species of the genus Halomonas are salt-tolerant organisms that have a versatile metabolism and can degrade a variety of xenobiotic compounds, utilizing them as their sole carbon source. In this study, we examined the genome of a Halomonas isolate from a hydrocarbon-contaminated site to search for chemosensory genes that might be responsible for the observed chemotactic behavior of this organism as well as for other responses to environmental cues. RESULTS: Using genome-wide comparative tools, our isolate was identified as a strain of Halomonas titanicae (strain KHS3), together with two other Halomonas strains with available genomes that had not been previously identified at a species level. The search for the main components of chemosensory pathways resulted in the identification of two clusters of chemosensory genes and a total of twenty-five chemoreceptor genes. One of the gene clusters is very similar to the che cluster from Escherichia coli and, presumably, it is responsible for the chemotactic behavior towards a variety of compounds. This gene cluster is present in 47 out of 56 analyzed Halomonas strains with available genomes. A second che-like cluster includes a gene coding for a diguanylate cyclase with a phosphotransfer and two receiver domains, as well as a gene coding for a chemoreceptor with a longer cytoplasmic domain than the other twenty-four. This seemingly independent pathway resembles the wsp pathway from Pseudomonas aeruginosa although it also presents several differences in gene order and domain composition. This second chemosensory gene cluster is only present in a sub-group within the genus Halomonas. Moreover, remarkably similar gene clusters are also found in some orders of Proteobacteria phylogenetically more distant from the Oceanospirillales, suggesting the occurrence of lateral transfer events. CONCLUSIONS: Chemosensory pathways were investigated within the genus Halomonas. A canonical chemotaxis pathway, controlled by a variable number of chemoreceptors, is widespread among Halomonas species. A second chemosensory pathway of unique organization that involves some type of c-di-GMP signaling was found to be present only in one branch of the genus, as well as in other proteobacterial lineages.
Asunto(s)
Proteínas Bacterianas/metabolismo , Halomonas/citología , Halomonas/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Quimiotaxis , Halomonas/genética , Modelos Moleculares , Filogenia , Conformación ProteicaRESUMEN
Mn-oxidizing potential of two metal-tolerant bacterial strains - Halomonas meridiana and Marinobacter algicola isolated from the South West Indian Ridge waters were compared at varying concentrations of Mn (II), i.e., 1, 10, and 100 µmol and mmol L-1 . Accompanying changes in their morphology and metabolism were also determined. At concentrations >1 mmol L-1 Mn (II), Mn-oxidizing potential of M. algicola was 2-7 times greater than that of H. meridiana. Scanning electron microscopy revealed that exposure to elevated metal content prompted bacterial cells especially those of M. algicola to been enveloped in exopolymeric material and form aggregates. Energy dispersive spectrometric analysis showed that exopolymeric material acts as a nucleation site for Mn deposition and oxide formation which occurs in the form of microspherical aggregates. These features show striking resemblance to biogenically produced Fe-Mn oxide deposits from Lau Basin. Surprisingly, diffractograms of auto-oxidized and bacterially formed Mn-oxide showed similarities to the hydrothermal vein mineral Rhodochrosite indicating that it can also be produced biotically. Elongation of cells by up to 4× the original size and distortion in cell shape were evident at Mn (II) concentrations >100 µmol L-1 . Marked differences in C-substrate utilization by the test strains were also observed in presence of Mn (II). A shift in use of substrates that are readily available in oceanic waters like N-acetyl-d-glucosamine to those that can be used under changing redox conditions (d-cellobiose) or in the presence of metal ions (d-arabinose, l-asparagine) were observed. These findings highlight the significant role of autochthonous bacteria in transforming reduced metal ions and aiding in the formation of metal oxides. Under natural or laboratory conditions, the mode of bacterially generated Mn-oxide tends to remain the same.
Asunto(s)
Halomonas/citología , Halomonas/metabolismo , Manganeso/metabolismo , Marinobacter/citología , Marinobacter/metabolismo , Agua de Mar/microbiología , Acetilglucosamina , Arabinosa , Asparagina , Celobiosa/metabolismo , Recuento de Colonia Microbiana , Genes Bacterianos/genética , Halomonas/clasificación , Halomonas/aislamiento & purificación , Océano Índico , Compuestos de Manganeso/química , Compuestos de Manganeso/metabolismo , Marinobacter/clasificación , Marinobacter/aislamiento & purificación , Metales/metabolismo , Microscopía Electrónica de Rastreo , Minerales/metabolismo , Océanos y Mares , Oxidación-Reducción , Óxidos/química , Óxidos/metabolismo , Filogenia , ARN Ribosómico 16S/genética , Agua de Mar/químicaRESUMEN
Research on halophilic microorganisms is important due to their relation to fundamental questions of survival of living organisms in a hostile environment. Here we introduce a novel method to stain halophiles with MitoTracker fluorescent dyes in their growth medium. The method is based on membrane-potential sensitive dyes, which were originally used to label mitochondria in eukaryotic cells. We demonstrate that these fluorescent dyes provide high staining efficiency and are beneficial for multi-staining purposes due to the spectral range covered (from orange to deep red). In contrast with other fluorescent dyes used so far, MitoTracker does not affect growth rate, and remains in cells after several washing steps and several generations in cell culture. The suggested dyes were tested on three archaeal (Hbt. salinarum, Haloferax sp., Halorubrum sp.) and two bacterial (Salicola sp., Halomonas sp.) strains of halophilic microorganisms. The new staining approach provides new insights into biology of Hbt. salinarum. We demonstrated the interconversion of rod-shaped cells of Hbt. salinarium to spheroplasts and submicron-sized spheres, as well as the cytoplasmic integrity of giant rod Hbt. salinarum species. By expanding the variety of tools available for halophile detection, MitoTracker dyes overcome long-standing limitations in fluorescence microscopy studies of halophiles.
Asunto(s)
Halobacteriaceae/citología , Halomonas/citología , Coloración y Etiquetado/métodos , Colorantes Fluorescentes/química , Potenciales de la Membrana , Microscopía FluorescenteRESUMEN
This study attempted to utilize Halomonas salina BCRC17875 to produce ectoine by optimizing the agitation speed and medium composition. In addition, the chemical structure of ectoine produced by H. salina BCRC17875 was determined. The results indicate that ectoine production reached 3.65 g/L at 38 h of cultivation when the agitation rate and NaCl concentration were fixed at 200 rpm and 2.0 M, respectively. It reached 9.20 g/L at 44 h of cultivation when the major medium components were yeast extract (56 g/L), glutamate (74.40 g/L), and ammonium sulfate (14 g/L). After the nitrogen concentration had been evaluated, evaluation of the nitrogen concentration revealed that the ectoine production reached 11.80 g/L at 44 h of cultivation when 56 g/L of yeast extract and 28 g/L of ammonium sulfate were used. Ectoine production reached 13.96 g/L at 44 h of cultivation when the carbon/nitrogen ratio was fixed at 3/1 using 84 g/L of yeast extract and 28 g/L of ammonium sulfate. Furthermore, the identification of ectoine were identified and characterized by fast atom bombardment mass spectrometry (FAB-MS) and 1H NMR. The results demonstrated a fermentation strategy was successful in increasing ectoine production, and that the fermentation medium of ectoine had commercialization potential.
Asunto(s)
Aminoácidos Diaminos/metabolismo , Técnicas de Cultivo Celular por Lotes/métodos , Halomonas/citología , Halomonas/metabolismo , Aminoácidos Diaminos/química , Técnicas Bacteriológicas/métodos , Carbono/metabolismo , Fermentación , Nitrógeno/metabolismo , Cloruro de Sodio/químicaRESUMEN
Studies on the halotolerance of bacteria are attractive to the fermentation industry. However, a lack of sufficient genomic information has precluded an investigation of the halotolerance of Halomonas beimenensis. Here, we describe the molecular mechanisms of saline adaptation in H. beimenensis based on high-throughput omics and Tn5 transposon mutagenesis. The H. beimenensis genome is 4.05 Mbp and contains 3,807 genes, which were sequenced using short and long reads obtained via deep sequencing. Sixteen Tn5 mutants with a loss of halotolerance were identified. Orthologs of the mutated genes, such as nqrA, trkA, atpC, nadA, and gdhB, have significant biological functions in sodium efflux, potassium uptake, hydrogen ion transport for energy conversion, and compatible solute synthesis, which are known to control halotolerance. Other genes, such as spoT, prkA, mtnN, rsbV, lon, smpB, rfbC, rfbP, tatB, acrR1, and lacA, function in cellular signaling, quorum sensing, transcription/translation, and cell motility also shown critical functions for promoting a halotolerance. In addition, KCl application increased halotolerance and potassium-dependent cell motility in a high-salinity environment. Our results demonstrated that a combination of omics and mutagenesis could be used to facilitate the mechanistic exploitation of saline adaptation in H. beimenensis, which can be applied for biotechnological purposes.
Asunto(s)
Adaptación Fisiológica , Elementos Transponibles de ADN/genética , Genómica/métodos , Halomonas/genética , Halomonas/fisiología , Mutagénesis/genética , Salinidad , Adaptación Fisiológica/efectos de los fármacos , Adaptación Fisiológica/genética , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Mapeo Cromosómico , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genoma Bacteriano , Halomonas/citología , Halomonas/crecimiento & desarrollo , Mutación/genética , Fenotipo , Filogenia , Potasio/farmacología , Transcriptoma/efectos de los fármacos , Transcriptoma/genéticaRESUMEN
In this study, a flow cytometry (FC) protocol was implemented to measure poly(3-hydroxybutyrate) (PHB) content in a halophilic bacterium, to have a faster and easier control of the process. The halophilic bacterium Halomonas boliviensis was stained with BODIPY 493/503 and analyzed using FC. Bacterial polymer accumulation induced by two different nutrient limitations during the operation of a 2 L bioreactor was studied using traditional gas chromatography (GC) analysis and FC. The application of this rapid and straightforward method is useful to obtain complex and precise information about PHB accumulation that could be used for the monitoring, control and optimization of the production of PHB. A clear correlation between the PHB concentration determined by GC and the fluorescence signal obtained from stained bacteria by using FC was observed. Additionally, the heterogeneity of bacterial population as a function of PHB content was measured. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:276-284, 2017.
Asunto(s)
Reactores Biológicos/microbiología , Cromatografía de Gases/métodos , Citometría de Flujo/métodos , Halomonas/citología , Halomonas/metabolismo , Hidroxibutiratos/metabolismo , Microscopía Fluorescente/métodos , Poliésteres/metabolismo , Compuestos de Boro , Colorantes Fluorescentes , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Biofouling, the colonization of artificial and natural surfaces by unwanted microorganisms, has an important economic impact on a wide range of industries. Low cost antifouling strategies are typically based on biocides which exhibit a negative environmental impact, affecting surrounding organisms related and not related to biofouling. Considering that the critical processes resulting in biofouling occur in the nanoscale/microscale dimensions, in this work we present a bionanotechnological approach to reduce adhesion of biofilm-producing bacteria Halomonas spp. CAM2 by introducing single layer graphene coatings. The use of this nanomaterial has been poorly explored for antifouling application. RESULTS: Our study revealed that graphene coatings modify material surface energy and electrostatic interaction between material and bacteria. Such nanoscale surface modification determine an important reduction over resulting bacterial adhesion and reduces the expression levels of genes related to adhesion when bacteria are in contact with graphene-coated material. CONCLUSIONS: Our results demonstrate that graphene coatings reduce considerably adhesion and expression levels of adhesion genes of biofilm-producing bacteria Halomonas spp. CAM2. Hydrophobic-hydrophilic interaction and repulsive electrostatic force dominate the interactions between Halomonas spp. CAM2 and material surface in saline media, impacting the final adhesion process. In addition no bactericide effect of graphene coatings was observed. The effect over biofilm formation is localized right at coated surface, in contrast to other antifouling techniques currently used, such as biocides.
Asunto(s)
Adhesión Bacteriana/efectos de los fármacos , Incrustaciones Biológicas , Grafito/farmacología , Halomonas/citología , Nanotecnología/métodos , Adhesinas Bacterianas/genética , Adhesinas Bacterianas/metabolismo , Secuencia de Aminoácidos , Materiales Biocompatibles Revestidos/farmacología , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Halomonas/ultraestructura , Interacciones Hidrofóbicas e Hidrofílicas , Microscopía Fluorescente , Datos de Secuencia Molecular , Nanoestructuras/química , Electricidad Estática , HumectabilidadRESUMEN
Several microorganisms produce polyhydroxyalkanoates (PHA). They are accumulated intracellularly as energy storage compounds. The PHAs are of interest because of their potential in biomedical applications. Halophilic bacteria and archaea are known to produce polyhydroxybutyrate (PHB). This paper describes production of a biodegradable copolymer, PHB-co-PHV by a moderately haloalkalitolerant Halomonas campisalis, isolated from Lonar Lake, India. The production of PHA was in the range of 45-81% on dry cell weight basis when the organism was grown in a production medium containing 1% (w/v) maltose and 0.1% (w/v) yeast extract, at pH ranging from 6 to 9 with an inoculum density of 10(5)-10(7) cells/ml of medium, for incubation period of 15-30 h and at 37 degrees C. The polymer produced by the organism is a hydroxyester with molecular weight of 1.3014 x 10(6). Its melting temperature was 171 degrees C. The (1)H NMR analysis revealed that the polymer was a copolymer of PHB-co-PHV. This could be achieved by providing simple carbon source viz. maltose.
Asunto(s)
Adaptación Fisiológica/efectos de los fármacos , Álcalis/farmacología , Agua Dulce/microbiología , Halomonas/aislamiento & purificación , Poliésteres/metabolismo , Sales (Química)/farmacología , Análisis de Varianza , Biodegradación Ambiental/efectos de los fármacos , Rastreo Diferencial de Calorimetría , Cromatografía de Gases , Cromatografía en Gel , Medios de Cultivo/farmacología , Halomonas/citología , Halomonas/efectos de los fármacos , Concentración de Iones de Hidrógeno/efectos de los fármacos , India , Espectroscopía de Resonancia Magnética , Peso Molecular , Estándares de Referencia , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Factores de TiempoRESUMEN
A moderately halophilic bacterium, strain CG2.1T, isolated from a solar saltern at Cabo de Gata, a wildlife reserve located in the province of Almería, southern Spain, was subjected to a polyphasic taxonomic study. This organism was an aerobic, motile, Gram-negative rod that produced orange-pigmented colonies. Strain CG2.1T was able to grow at salinities of 3-25 % (w/v) and at temperatures of 15-40 degrees C. The pH range for growth was 5-9. Strain CG2.1T was a heterotroph capable of utilizing various carbohydrates as carbon sources. The organism reduced nitrate and showed phenylalanine deaminase activity. The major fatty acids were C(18 : 1)omega7c, C(16 : 0) and C(19 : 0) cyclo omega8c. The DNA G+C content was 60.9 mol%. On the basis of the phenotypic and phylogenetic data, strain CG2.1T appeared to be a member of the genus Halomonas and clustered closely with Halomonas marisflavi (97.1 % 16S rRNA gene sequence similarity). However, the level of DNA-DNA relatedness between the novel isolate and the most closely related Halomonas species was low. On the basis of these data, strain CG2.1T represents a novel member of the genus Halomonas, for which the name Halomonas indalinina is proposed. The type strain is CG2.1T (=CECT 5902T=LMG 23625T).
Asunto(s)
Halomonas/clasificación , Halomonas/aislamiento & purificación , Agua de Mar/microbiología , Aerobiosis , Aminoácido Oxidorreductasas/análisis , Antibacterianos/farmacología , Técnicas de Tipificación Bacteriana , Composición de Base , Metabolismo de los Hidratos de Carbono , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Ácidos Grasos/química , Genes de ARNr , Halomonas/citología , Halomonas/fisiología , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Movimiento , Nitratos/metabolismo , Oxidación-Reducción , Filogenia , Pigmentos Biológicos/biosíntesis , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Solución Salina Hipertónica/farmacología , España , Temperatura , Microbiología del AguaRESUMEN
Vitamin B12 (cobalamin) was identified nearly 80 years ago as the anti-pernicious anaemia factor in liver, and its importance in human health and disease has resulted in much work on its uptake, cellular transport and utilization. Plants do not contain cobalamin because they have no cobalamin-dependent enzymes. Deficiencies are therefore common in strict vegetarians, and in the elderly, who are susceptible to an autoimmune disorder that prevents its efficient uptake. In contrast, many algae are rich in vitamin B12, with some species, such as Porphyra yezoensis (Nori), containing as much cobalamin as liver. Despite this, the role of the cofactor in algal metabolism remains unknown, as does the source of the vitamin for these organisms. A survey of 326 algal species revealed that 171 species require exogenous vitamin B12 for growth, implying that more than half of the algal kingdom are cobalamin auxotrophs. Here we show that the role of vitamin B12 in algal metabolism is primarily as a cofactor for vitamin B12-dependent methionine synthase, and that cobalamin auxotrophy has arisen numerous times throughout evolution, probably owing to the loss of the vitamin B12-independent form of the enzyme. The source of cobalamin seems to be bacteria, indicating an important and unsuspected symbiosis.
Asunto(s)
Bacterias/metabolismo , Eucariontes/metabolismo , Simbiosis , Vitamina B 12/metabolismo , 5-Metiltetrahidrofolato-Homocisteína S-Metiltransferasa/metabolismo , Bacterias/citología , Bacterias/crecimiento & desarrollo , Técnicas de Cocultivo , Eucariontes/clasificación , Eucariontes/citología , Eucariontes/genética , Genoma , Genómica , Halomonas/citología , Halomonas/crecimiento & desarrollo , Halomonas/metabolismoRESUMEN
The moderately halophilic strain Halomonas maura S-30 produces a high-molecular-mass acidic polymer (4.7 x 10(6) Da) composed of repeating units of mannose, galactose, glucose and glucuronic acid. This exopolysaccharide (EPS), known as mauran, has interesting functional properties that make it suitable for use in many industrial fields. Analysis of the flanking regions of a mini-Tn5 insertion site in an EPS-deficient mutant of H. maura, strain TK71, led to the identification of five ORFs (epsABCDJ), which form part of a gene cluster (eps) with the same structural organization as others involved in the biosynthesis of group 1 capsules and some EPSs. Conserved genetic features were found such as JUMPstart and ops elements, which are characteristically located preceding the gene clusters for bacterial polysaccharides. On the basis of their amino-acid-sequence homologies, their putative hydropathy profiles and the effect of their mutations, it is predicted that EpsA (an exporter-protein homologue belonging to the OMA family) and EpsC (a chain-length-regulator homologue belonging to the PCP family) play a role in the assembly, polymerization and translocation of mauran. The possibility that mauran might be synthesized via a Wzy-like biosynthesis system, just as it is for many other polysaccharides, is also discussed. This hypothesis is supported by the fact that EpsJ is homologous with some members of the PST-exporter-protein family, which seems to function together with each OMA-PCP pair in polysaccharide transport in Gram-negative bacteria, transferring the assembled lipid-linked repeating units from the cytoplasmic membrane to the periplasmic space. Maximum induction of the eps genes is reached during stationary phase in the presence of 5 % (w/v) marine salts.
Asunto(s)
Proteínas Bacterianas/metabolismo , Halomonas/metabolismo , Familia de Multigenes/fisiología , Polisacáridos Bacterianos/biosíntesis , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Halomonas/citología , Halomonas/genética , Datos de Secuencia Molecular , Polisacáridos Bacterianos/químicaRESUMEN
Some members of the moderately halophilic genus Halomonas, such as H. eurihalina, H. maura, H. ventosae and H. anticariensis, produce exopolysaccharides with applications in many industrial fields. We report here that these four species also produce autoinducer molecules that are involved in the cell-to-cell signaling process known as quorum sensing. By using the N-acyl homoserine lactone (AHL) indicator strains Agrobacterium tumefaciens NTL4 (pZRL4) and Chromobacterium violaceum CV026, we discovered that all the Halomonas strains examined synthesize detectable AHL signal molecules. The synthesis of these compounds was growth-phase dependent and maximal activity was reached during the late exponential to stationary phases. One of these AHLs seems to be synthesized only in the stationary phase. Some of the AHLs produced by H. anticariens FP35(T) were identified by gas chromatography/mass spectrometry and electrospray ionization tandem mass spectrometry as N-butanoyl homoserine lactone (C(4)-HL), N-hexanoyl homoserine lactone (C(6)-HL), N-octanoyl homoserine lactone (C(8)-HL) and N-dodecanoyl homoserine lactone (C(12)-HL). This study suggests that quorum sensing may also play an important role in extreme environments.
Asunto(s)
4-Butirolactona/análogos & derivados , Halomonas/química , Halomonas/metabolismo , Polisacáridos/biosíntesis , 4-Butirolactona/análisis , 4-Butirolactona/biosíntesis , 4-Butirolactona/química , División Celular , Halomonas/clasificación , Halomonas/citología , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
A moderately halophilic bacterium, strain SS20(T), capable of growing at salinities of 1-20 % (w/v) NaCl was isolated from a solar saltern of the Dangjin area in Korea and was characterized taxonomically. Strain SS20(T) was a Gram-negative bacterium comprising motile, short rods. Its major cellular fatty acids were C(18 : 1)omega7c, C(19 : 0)omega8c cyclo and C(16 : 0). The DNA G+C content was 70 mol% and the predominant ubiquinone was Q-9. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SS20(T) belonged to the genus Halomonas. The levels of 16S rRNA gene sequence similarity to the type strains of Halomonas species were in the range 93.0-97.5 %. The levels of DNA-DNA relatedness between strain SS20(T) and the type strains of phylogenetically closely related Halomonas species were in the range 5.3-12.3 %. On the basis of physiological and molecular properties, strain SS20(T) represents a novel species of the genus Halomonas, for which the name Halomonas koreensis sp. nov. is proposed. The type strain is SS20(T) (=KCTC 12127(T)=JCM 12237(T)).
Asunto(s)
Sedimentos Geológicos/microbiología , Halomonas/clasificación , Halomonas/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/química , ADN Bacteriano/aislamiento & purificación , ADN Ribosómico/química , ADN Ribosómico/aislamiento & purificación , Ácidos Grasos/análisis , Ácidos Grasos/aislamiento & purificación , Genes de ARNr , Violeta de Genciana , Halomonas/citología , Halomonas/fisiología , Corea (Geográfico) , Datos de Secuencia Molecular , Movimiento , Hibridación de Ácido Nucleico , Fenazinas , Filogenia , Quinonas/análisis , Quinonas/aislamiento & purificación , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Agua de Mar/microbiología , Análisis de Secuencia de ADNRESUMEN
Mauran is an anionic, sulfated heteropolysaccharide with a high uronic-acid content, synthesized by strain S-30 of the halophilic bacterium Halomonas maura. Under optimum environmental and nutritional conditions, it is capable of producing up to 3.8 g of mauran per liter of medium. Aqueous solutions of mauran are highly viscous and display pseudoplastic, viscoelastic and thixotropic behavior. Its viscosity is stable over a wide pH range (3-11), after freezing-thawing processes, and in the presence of sucrose, salts, surfactants and alpha-hydroxyl acids. It has a high capacity for binding lead and other cations. Its molecular mass when collected from an MY medium supplemented with 2.5% w/v salt during the stationary growth phase is 4.7x10(6) Da.
Asunto(s)
Biotecnología , Halomonas/metabolismo , Polisacáridos Bacterianos/biosíntesis , Polisacáridos Bacterianos/química , Microbiología del Suelo , Medios de Cultivo , Halomonas/citología , Halomonas/genética , Concentración de Iones de Hidrógeno , Intercambio Iónico , Espectrometría de Masas , Metales/química , Microscopía Electrónica , Peso Molecular , ViscosidadRESUMEN
Halomonas eurihalina strain H-28 is a moderately halophilic bacterium that produces an extracellular polysaccharide not only in media with glucose but also in media supplemented with hydrocarbons (n-tetradecane, n-hexadecane, n-octane, xylene, mineral light oil, mineral heavy oil, petrol, or crude oil). In this study we investigated yield production, chemical composition, viscosity, and emulsifying activity of exopolysaccharides (EPS) extracted from the different media used. The largest amounts of biopolymer were synthesized in media with glucose and n-hexadecane. Chemical composition varied with culture conditions; thus EPS from cultures grown in the presence of hydrocarbons had lower contents of carbohydrates and proteins than EPS from media with glucose. However, the percentages of uronic acids, acetyls, and sulfates were always higher than glucose EPS. Crude oil was the substrate most effectively emulsified. All EPS were capable of emulsifying crude oil more efficiently than the three control surfactants tested (Tween 20, Tween 80, and Triton X-100). All polymers gave low viscosity solutions. EPS H28 could be attractive for application in the oil industry and/or in bioremediation processes, bearing in mind not only its functional properties, but also the capacity of producer strain H-28 to grow in the presence of high salt concentrations and oil substrates.