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1.
PLoS Genet ; 19(3): e1010672, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36930675

RESUMEN

Bacterial genome diversity is influenced by prophages, which are viral genomes integrated into the bacterial chromosome. Most prophage genes are silent but those that are expressed can provide unexpected properties to their host. Using as a model E. coli K-12 that carries 9 defective prophages in its genome, we aimed at highlighting the impact of genes encoded by prophages on host physiology. We focused our work on AppY, a transcriptional regulator encoded on the DLP12 prophage. By performing RNA-Seq experiments, we showed that AppY production modulates the expression of more than 200 genes. Among them, 11 were identified by ChIP-Seq as direct AppY targets. AppY directly and positively regulates several genes involved in the acid stress response including the master regulator gene gadE but also nhaR and gadY, two genes important for biofilm formation. Moreover, AppY indirectly and negatively impacts bacterial motility by favoring the degradation of FlhDC, the master regulator of the flagella biosynthesis. As a consequence of these regulatory effects, AppY increases acid stress resistance and biofilm formation while also causing a strong defect in motility. Our research shed light on the importance to consider the genetic interactions occurring between prophages and bacteria to fully understand bacterial physiology. It also highlights how a prophage-encoded transcriptional regulator integrates in a complex manner into the host regulatory network and how it benefits its host, allowing it to cope with changing environmental conditions.


Asunto(s)
Proteínas de Escherichia coli , Escherichia coli , Escherichia coli/genética , Escherichia coli/metabolismo , Profagos/genética , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Genoma Bacteriano/genética , Bacterias/genética
2.
Int J Mol Sci ; 23(16)2022 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-36012174

RESUMEN

Salmonella enterica, a Gram-negative zoonotic bacterium, is mainly a food-borne pathogen and the main cause of diarrhea in humans worldwide. The main reservoirs are found in poultry farms, but they are also found in wild birds. The development of antibiotic resistance in S. enterica species raises concerns about the future of efficient therapies against this pathogen and revives the interest in bacteriophages as a useful therapy against bacterial infections. Here, we aimed to decipher and functionally annotate 10 new Salmonella phage genomes isolated in Spain in the light of phage therapy. We designed a bioinformatic pipeline using available building blocks to de novo assemble genomes and perform syntaxic annotation. We then used genome-wide analyses for taxonomic annotation enabled by vContact2 and VICTOR. We were also particularly interested in improving functional annotation using remote homologies detection and comparisons with the recently published phage-specific PHROG protein database. Finally, we searched for useful functions for phage therapy, such as systems encoded by the phage to circumvent cellular defenses with a particular focus on anti-CRISPR proteins. We, thus, were able to genetically characterize nine virulent phages and one temperate phage and identify putative functions relevant to the formulation of phage cocktails for Salmonella biocontrol.


Asunto(s)
Bacteriófagos , Terapia de Fagos , Salmonelosis Animal , Fagos de Salmonella , Salmonella enterica , Animales , Bacteriófagos/genética , Estudio de Asociación del Genoma Completo , Humanos , Fagos de Salmonella/genética , Salmonella enterica/genética
3.
Environ Microbiol ; 20(12): 4415-4430, 2018 12.
Artículo en Inglés | MEDLINE | ID: mdl-30043533

RESUMEN

Ecological and evolutionary processes involved in magnetotactic bacteria (MTB) adaptation to their environment have been a matter of debate for many years. Ongoing efforts for their characterization are progressively contributing to understand these processes, including the genetic and molecular mechanisms responsible for biomineralization. Despite numerous culture-independent MTB characterizations, essentially within the Proteobacteria phylum, only few species have been isolated in culture because of their complex growth conditions. Here, we report a newly cultivated magnetotactic, microaerophilic and chemoorganoheterotrophic bacterium isolated from the Mediterranean Sea in Marseille, France: Candidatus Terasakiella magnetica strain PR-1 that belongs to an Alphaproteobacteria genus with no magnetotactic relative. By comparing the morphology and the whole genome shotgun sequence of this MTB with those of closer relatives, we brought further evidence that the apparent vertical ancestry of magnetosome genes suggested by previous studies within Alphaproteobacteria hides a more complex evolutionary history involving horizontal gene transfers and/or duplication events before and after the emergence of Magnetospirillum, Magnetovibrio and Magnetospira genera. A genome-scale comparative genomics analysis identified several additional candidate functions and genes that could be specifically associated to MTB lifestyle in this class of bacteria.


Asunto(s)
Alphaproteobacteria/genética , Evolución Molecular , Magnetosomas/genética , Francia , Transferencia de Gen Horizontal , Genoma Bacteriano , Magnetismo , Mar Mediterráneo , Microbiología del Agua
4.
J Bacteriol ; 196(17): 3111-21, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24957623

RESUMEN

Many bacterial species contain multiple actin-like proteins tasked with the execution of crucial cell biological functions. MamK, an actin-like protein found in magnetotactic bacteria, is important in organizing magnetosome organelles into chains that are used for navigation along geomagnetic fields. MamK and numerous other magnetosome formation factors are encoded by a genetic island termed the magnetosome island. Unlike most magnetotactic bacteria, Magnetospirillum magneticum AMB-1 (AMB-1) contains a second island of magnetosome-related genes that was named the magnetosome islet. A homologous copy of mamK, mamK-like, resides within this islet and encodes a protein capable of filament formation in vitro. Previous work had shown that mamK-like is expressed in vivo, but its function, if any, had remained unknown. Though MamK-like is highly similar to MamK, it contains a mutation that in MamK and other actins blocks ATPase activity in vitro and filament dynamics in vivo. Here, using genetic analysis, we demonstrate that mamK-like has an in vivo role in assisting organelle alignment. In addition, MamK-like forms filaments in vivo in a manner that is dependent on the presence of MamK and the two proteins interact in a yeast two-hybrid assay. Surprisingly, despite the ATPase active-site mutation, MamK-like is capable of ATP hydrolysis in vitro and promotes MamK filament turnover in vivo. Taken together, these experiments suggest that direct interactions between MamK and MamK-like contribute to magnetosome alignment in AMB-1.


Asunto(s)
Actinas/química , Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Magnetosomas/fisiología , Magnetospirillum/metabolismo , Adenosina Trifosfatasas/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Magnetospirillum/citología , Magnetospirillum/genética , Datos de Secuencia Molecular , Mutación
5.
Environ Microbiol ; 16(2): 525-44, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23841906

RESUMEN

Magnetotactic bacteria (MTB) are capable of synthesizing intracellular organelles, the magnetosomes, that are membrane-bounded magnetite or greigite crystals arranged in chains. Although MTB are widely spread in various ecosystems, few axenic cultures are available, and only freshwater Magnetospirillum spp. have been genetically analysed. Here, we present the complete genome sequence of a marine magnetotactic spirillum, Magnetospira sp. QH-2. The high number of repeats and transposable elements account for the differences in QH-2 genome structure compared with other relatives. Gene cluster synteny and gene correlation analyses indicate that the insertion of the magnetosome island in the QH-2 genome occurred after divergence between freshwater and marine magnetospirilla. The presence of a sodium-quinone reductase, sodium transporters and other functional genes are evidence of the adaptive evolution of Magnetospira sp. QH-2 to the marine ecosystem. Genes well conserved among freshwater magnetospirilla for nitrogen fixation and assimilatory nitrate respiration are absent from the QH-2 genome. Unlike freshwater Magnetospirillum spp., marine Magnetospira sp. QH-2 neither has TonB and TonB-dependent receptors nor does it grow on trace amounts of iron. Taken together, our results show a distinct, adaptive evolution of Magnetospira sp. QH-2 to marine sediments in comparison with its closely related freshwater counterparts.


Asunto(s)
Evolución Biológica , Ecosistema , Genoma Bacteriano , Magnetospirillum/genética , Adaptación Biológica/genética , Proteínas Bacterianas/genética , Hibridación Genómica Comparativa , Elementos Transponibles de ADN , ADN Bacteriano/genética , Islas Genómicas , Magnetosomas/genética , Magnetospirillum/fisiología , Familia de Multigenes , Filogenia , Quinona Reductasas/genética , Agua de Mar/microbiología , Simportadores/genética , Sintenía
6.
Environ Microbiol ; 15(10): 2712-35, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23607663

RESUMEN

Magnetotactic bacteria (MTB) represent a group of diverse motile prokaryotes that biomineralize magnetosomes, the organelles responsible for magnetotaxis. Magnetosomes consist of intracellular, membrane-bounded, tens-of-nanometre-sized crystals of the magnetic minerals magnetite (Fe3O4) or greigite (Fe3S4) and are usually organized as a chain within the cell acting like a compass needle. Most information regarding the biomineralization processes involved in magnetosome formation comes from studies involving Alphaproteobacteria species which biomineralize cuboctahedral and elongated prismatic crystals of magnetite. Many magnetosome genes, the mam genes, identified in these organisms are conserved in all known MTB. Here we present a comparative genomic analysis of magnetotactic Deltaproteobacteria that synthesize bullet-shaped crystals of magnetite and/or greigite. We show that in addition to mam genes, there is a conserved set of genes, designated mad genes, specific to the magnetotactic Deltaproteobacteria, some also being present in Candidatus Magnetobacterium bavaricum of the Nitrospirae phylum, but absent in the magnetotactic Alphaproteobacteria. Our results suggest that the number of genes associated with magnetotaxis in magnetotactic Deltaproteobacteria is larger than previously thought. We also demonstrate that the minimum set of mam genes necessary for magnetosome formation in Magnetospirillum is also conserved in magnetite-producing, magnetotactic Deltaproteobacteria. Some putative novel functions of mad genes are discussed.


Asunto(s)
Deltaproteobacteria/genética , Óxido Ferrosoférrico , Genoma Bacteriano/genética , Hierro , Magnetosomas/genética , Sulfuros , Secuencia Conservada , Deltaproteobacteria/clasificación , Magnetismo , Datos de Secuencia Molecular , Familia de Multigenes/genética , Filogenia
7.
Environ Microbiol ; 15(8): 2267-74, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23438345

RESUMEN

Horizontal gene transfer (HGT), the transfer of genetic material other than by descent, is thought to have played significant roles in the evolution and distribution of genes in prokaryotes. These include those responsible for the ability of motile, aquatic magnetotactic bacteria (MTB) to align and swim along magnetic field lines and the biomineralization of magnetosomes that are responsible for this behaviour. There is some genomic evidence that HGT might be responsible for the distribution of magnetosome genes in different phylogenetic groups of bacteria. For example, in the genomes of a number of MTB, magnetosome genes are present as clusters within a larger structure known as the magnetosome genomic island surrounded by mobile elements such as insertion sequences and transposases as well as tRNA genes. Despite this, there is no strong direct proof of HGT between these organisms. Here we show that a phylogenetic tree based on magnetosome protein amino acid sequences from a number of MTB was congruent with the tree based on the organisms' 16S rRNA gene sequences. This shows that evolution and divergence of these proteins and the 16S rRNA gene occurred similarly. This suggests that magnetotaxis originated monophyletically in the Proteobacteria phylum and implies that the common ancestor of all Proteobacteria was magnetotactic.


Asunto(s)
Bacterias/clasificación , Bacterias/genética , Magnetosomas/genética , Filogenia , Bacterias/metabolismo , Secuencia de Bases , Elementos Transponibles de ADN/genética , Transferencia de Gen Horizontal/genética , Genómica , Proteobacteria/clasificación , Proteobacteria/genética , Proteobacteria/metabolismo , ARN Ribosómico 16S/genética
8.
Biochem Soc Trans ; 40(6): 1319-23, 2012 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-23176475

RESUMEN

Magnetotactic bacteria consist of a group of taxonomically, physiologically and morphologically diverse prokaryotes, with the singular ability to align with geomagnetic field lines, a phenomenon referred to as magnetotaxis. This magnetotactic property is due to the presence of iron-rich crystals embedded in lipidic vesicles forming an organelle called the magnetosome. Magnetosomes are composed of single-magnetic-domain nanocrystals of magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4)) embedded in biological membranes, thereby forming a prokaryotic organelle. Four specific steps are described in this organelle formation: (i) membrane specialization, (ii) iron acquisition, (iii) magnetite (or greigite) biocrystallization, and (iv) magnetosome alignment. The formation of these magnetic crystals is a genetically controlled process, which is governed by enzyme-catalysed processes. On the basis of protein sequence analysis of genes known to be involved in magnetosome formation in Magnetospirillum magneticum AMB-1, we have identified a subset of three membrane-associated or periplasmic proteins containing a double cytochrome c signature motif CXXCH: MamE, MamP and MamT. The presence of these proteins suggests the existence of an electron-transport chain inside the magnetosome, contributing to the process of biocrystallization. We have performed heterologous expression in E. coli of the cytochrome c motif-containing domains of MamE, MamP and MamT. Initial biophysical characterization has confirmed that MamE, MamP and MamT are indeed c-type cytochromes. Furthermore, determination of redox potentials for this new family of c-type cytochromes reveals midpoint potentials of -76 and -32 mV for MamP and MamE respectively.


Asunto(s)
Magnetosomas/metabolismo , Magnetospirillum/genética , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Cristalización , Grupo Citocromo c/química , Grupo Citocromo c/genética , Grupo Citocromo c/metabolismo , Transporte de Electrón , Magnetosomas/genética , Magnetosomas/fisiología , Magnetospirillum/metabolismo , Magnetospirillum/fisiología , Datos de Secuencia Molecular , Oxidación-Reducción , Estructura Terciaria de Proteína
9.
Biochim Biophys Acta ; 1797(11): 1780-94, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-20655292

RESUMEN

Photosynthetic membranes accommodate densely packed light-harvesting complexes which absorb light and convey excitation to the reaction center (RC). The relationship between the fluorescence yield (phi) and the fraction (x) of closed RCs is informative about the probability for an excitation reaching a closed RC to be redirected to another RC. In this work, we have examined in this respect membranes from various bacteria and searched for a correlation with the arrangement of the light-harvesting complexes as known from atomic force or electron microscopies. A first part of the paper is devoted to a theoretical study analyzing the phi(x) relationship in various models: monomeric or dimeric RC-LH1 core complexes, with or without the peripheral LH2 complexes. We show that the simple "homogeneous" kinetic treatment used here agrees well with more detailed master equation calculations. We also discuss the agreement between information derived from the present technique and from singlet annihilation experiments. The experimental results show that the enhancement of the cross section of open RCs due to excitation transfer from closed units varies from 1.5 to 3 depending on species. The ratio of the core to core transfer rate (including the indirect pathway via LH2) to the rate of trapping in open units is in the range of 0.5 to 4. It is about 1 in Rhodobacter sphaeroides and does not increase significantly in mutants lacking LH2-despite the more numerous contacts between the dimeric core complexes expected in this case. The connectivity in this bacterium is due in good part to the fast transfer between the two partners of the dimeric (RC-LH1-PufX)(2) complex. The connectivity is however increased in the carotenoidless and LH2-less strain R26, which we ascribe to an anomalous LH1. A relatively high connectivity was found in Rhodospirillum photometricum, although not as high as predicted in the calculations of Fassioli et al. (2010). This illustrates a more general discrepancy between the measured efficiency of core to core excitation transfer and theoretical estimates. We argue that the limited core to core connectivity found in purple bacteria may reflect a trade-off between light-harvesting efficiency and the hindrance to quinone diffusion that would result from too tightly packed LH complexes.


Asunto(s)
Complejos de Proteína Captadores de Luz/metabolismo , Modelos Teóricos , Fotosíntesis , Proteobacteria/química , Proteobacteria/metabolismo , Membrana Celular/metabolismo , Difusión , Cinética , Luz , Proteobacteria/genética , Quinonas/metabolismo
10.
Viruses ; 13(5)2021 04 21.
Artículo en Inglés | MEDLINE | ID: mdl-33919362

RESUMEN

Xylella fastidiosa (Xf) is a plant pathogen causing significant losses in agriculture worldwide. Originating from America, this bacterium caused recent epidemics in southern Europe and is thus considered an emerging pathogen. As the European regulations do not authorize antibiotic treatment in plants, alternative treatments are urgently needed to control the spread of the pathogen and eventually to cure infected crops. One such alternative is the use of phage therapy, developed more than 100 years ago to cure human dysentery and nowadays adapted to agriculture. The first step towards phage therapy is the isolation of the appropriate bacteriophages. With this goal, we searched for phages able to infect Xf strains that are endemic in the Mediterranean area. However, as Xf is truly a fastidious organism, we chose the phylogenetically closest and relatively fast-growing organism X. albineans as a surrogate host for the isolation step. Our results showed the isolation from various sources and preliminary characterization of several phages active on different Xf strains, namely, from the fastidiosa (Xff), multiplex (Xfm), and pauca (Xfp) subspecies, as well as on X. albilineans. We sequenced their genomes, described their genomic features, and provided a phylogeny analysis that allowed us to propose new taxonomic elements. Among the 14 genomes sequenced, we could identify two new phage species, belonging to two new genera of the Caudoviricetes order, namely, Usmevirus (Podoviridae family) and Subavirus (Siphoviridae family). Interestingly, no specific phages could be isolated from infected plant samples, whereas one was isolated from vector insects captured in a contaminated area, and several from surface and sewage waters from the Marseille area.


Asunto(s)
Bacteriófagos/fisiología , Plantas/microbiología , Xanthomonas/virología , Xylella/virología , Bacteriófagos/clasificación , Bacteriófagos/aislamiento & purificación , Bacteriófagos/ultraestructura , ADN Viral , Especificidad del Huésped , Filogenia , Enfermedades de las Plantas/microbiología , Tropismo Viral , Virulencia , Xanthomonas/aislamiento & purificación , Xylella/aislamiento & purificación
11.
ISME J ; 15(1): 1-18, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32839547

RESUMEN

Bacteria synthesize a wide range of intracellular submicrometer-sized inorganic precipitates of diverse chemical compositions and structures, called biominerals. Their occurrences, functions and ultrastructures are not yet fully described despite great advances in our knowledge of microbial diversity. Here, we report bacteria inhabiting the sediments and water column of the permanently stratified ferruginous Lake Pavin, that have the peculiarity to biomineralize both intracellular magnetic particles and calcium carbonate granules. Based on an ultrastructural characterization using transmission electron microscopy (TEM) and synchrotron-based scanning transmission X-ray microscopy (STXM), we showed that the calcium carbonate granules are amorphous and contained within membrane-delimited vesicles. Single-cell sorting, correlative fluorescent in situ hybridization (FISH), scanning electron microscopy (SEM) and molecular typing of populations inhabiting sediments affiliated these bacteria to a new genus of the Alphaproteobacteria. The partially assembled genome sequence of a representative isolate revealed an atypical structure of the magnetosome gene cluster while geochemical analyses indicate that calcium carbonate production is an active process that costs energy to the cell to maintain an environment suitable for their formation. This discovery further expands the diversity of organisms capable of intracellular Ca-carbonate biomineralization. If the role of such biomineralization is still unclear, cell behaviour suggests that it may participate to cell motility in aquatic habitats as magnetite biomineralization does.


Asunto(s)
Alphaproteobacteria , Magnetosomas , Alphaproteobacteria/genética , Biomineralización , Carbonatos , Óxido Ferrosoférrico , Hibridación Fluorescente in Situ
12.
ISME J ; 14(7): 1783-1794, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32296121

RESUMEN

Under the same selection pressures, two genetically divergent populations may evolve in parallel toward the same adaptive solutions. Here, we hypothesized that magnetotaxis (i.e., magnetically guided chemotaxis) represents a key adaptation to micro-oxic habitats in aquatic sediments and that its parallel evolution homogenized the phenotypes of two evolutionary divergent clusters of freshwater spirilla. All magnetotactic bacteria affiliated to the Magnetospirillum genus (Alphaproteobacteria class) biomineralize the same magnetic particle chains and share highly similar physiological and ultrastructural features. We looked for the processes that could have contributed at shaping such an evolutionary pattern by reconciling species and gene trees using newly sequenced genomes of Magnetospirillum related bacteria. We showed that repeated horizontal gene transfers and homologous recombination of entire operons contributed to the parallel evolution of magnetotaxis. We propose that such processes could represent a more parsimonious and rapid solution for adaptation compared with independent and repeated de novo mutations, especially in the case of traits as complex as magnetotaxis involving tens of interacting proteins. Besides strengthening the idea about the importance of such a function in micro-oxic habitats, these results reinforce previous observations in experimental evolution suggesting that gene flow could alleviate clonal interference and speed up adaptation under some circumstances.


Asunto(s)
Alphaproteobacteria , Magnetospirillum , Bacterias/genética , Transferencia de Gen Horizontal , Bacterias Gramnegativas , Magnetospirillum/genética
13.
J Am Chem Soc ; 130(29): 9318-31, 2008 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-18588291

RESUMEN

The competition between the P(+)Q(A)(-) --> PQ(A) charge recombination (P, bacteriochlorophyll pair acting as primary photochemical electron donor) and the electron transfer to the secondary quinone acceptor Q(A)(-)Q(B) --> Q(A)Q(B)(-) (Q(A) and Q(B), primary and secondary electron accepting quinones) was investigated in chromatophores of Rb. capsulatus, varying the temperature down to -65 degrees C. The analysis of the flash-induced pattern for the formation of P(+)Q(A)Q(B)(-) shows that the diminished yield, when lowering the temperature, is not due to a homogeneous slowing of the rate constant k(AB) of the Q(A)(-)Q(B) --> Q(A)Q(B)(-) electron transfer but to a distribution of conformations that modulate the electron transfer rate over more than 3 orders of magnitude. This distribution appears "frozen", as no dynamic redistribution was observed over time ranges > 10 s (below -25 degrees C). The kinetic pattern was analyzed to estimate the shape of the distribution of k(AB), showing a bell-shaped band on the high rate side and a fraction of "blocked" reaction centers (RCs) with very slow k(AB). When the temperature is lowered, the high rate band moves to slower rate regions and the fraction of blocked RCs increases at the expense of the high rate band. The RCs that recombine from the P(+)Q(A)Q(B)(-) state appear temporarily converted to a state with rapid k(AB), indicating that the stabilized state described by Kleinfeld et al. (Biochemistry 1984, 23, 5780-5786) is still accessible at -60 degrees C.


Asunto(s)
Bacterioclorofilas/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Bacterioclorofilas/metabolismo , Benzoquinonas/química , Benzoquinonas/metabolismo , Frío , Citocromos c2/química , Citocromos c2/metabolismo , Electrones , Cinética , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Conformación Proteica , Rhodobacter capsulatus/química , Rhodobacter capsulatus/metabolismo , Espectrometría de Fluorescencia , Termodinámica , Ubiquinona/química , Ubiquinona/metabolismo
14.
Environ Sci Pollut Res Int ; 24(1): 25-32, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26769474

RESUMEN

Whole-cell biosensors based on the reporter gene system can offer rapid detection of trace levels of organic or metallic compounds in water. They are well characterized in laboratory conditions, but their transfer into technological devices for the surveillance of water networks remains at a conceptual level. The development of a semi-autonomous inline water analyzer stumbles across the conservation of the bacterial biosensors over a period of time compatible with the autonomy requested by the end-user while maintaining a satisfactory sensitivity, specificity, and time response. We focused here on assessing the effect of lyophilization on two biosensors based on the reporter gene system and hosted in Escherichia coli. The reporter gene used here is the entire bacterial luciferase lux operon (luxCDABE) for an autonomous bioluminescence emission without the need to add any substrate. In the cell-survival biosensor that is used to determine the overall fitness of the bacteria when mixed with the water sample, lux expression is driven by a constitutive E. coli promoter PrpoD. In the arsenite biosensor, the arsenite-inducible promoter P ars involved in arsenite resistance in E. coli controls lux expression. Evaluation of the shelf life of these lyophilized biosensors kept at 4 °C over a year evidenced that about 40 % of the lyophilized cells can be revived in such storage conditions. The performances of the lyophilized biosensor after 7 months in storage are maintained, with a detection limit of 0.2 µM arsenite for a response in about an hour with good reproducibility. These results pave the way to the use in tandem of both biosensors (one for general toxicity and one for arsenite contamination) as consumables of an autonomous analyzer in the field.


Asunto(s)
Arsenitos/química , Técnicas Biosensibles/instrumentación , Escherichia coli/metabolismo , Luciferasas de la Bacteria/metabolismo , Escherichia coli/genética , Regulación Bacteriana de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Genes Reporteros , Luciferasas de la Bacteria/genética , Operón , Regiones Promotoras Genéticas , Reproducibilidad de los Resultados
15.
Viruses ; 9(4)2017 04 21.
Artículo en Inglés | MEDLINE | ID: mdl-28430166

RESUMEN

The study of bacteriophages (viruses of bacteria) includes a variety of approaches, such as structural biology, genetics, ecology, and evolution, with increasingly important implications for therapeutic and industrial uses. Researchers working with phages in France have recently established a network to facilitate the exchange on complementary approaches, but also to engage new collaborations. Here, we provide a summary of the topics presented during the second meeting of the French Phage Network that took place in Marseille in November 2016.


Asunto(s)
Bacteriófagos/genética , Bacteriófagos/fisiología , Investigación Biomédica/organización & administración , Redes Comunitarias , Francia , Colaboración Intersectorial
16.
Syst Appl Microbiol ; 40(5): 280-289, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28622795

RESUMEN

A magnetotactic bacterium, designated strain BW-1T, was isolated from a brackish spring in Death Valley National Park (California, USA) and cultivated in axenic culture. The Gram-negative cells of strain BW-1T are relatively large and rod-shaped and possess a single polar flagellum (monotrichous). This strain is the first magnetotactic bacterium isolated in axenic culture capable of producing greigite and/or magnetite nanocrystals aligned in one or more chains per cell. Strain BW-1T is an obligate anaerobe that grows chemoorganoheterotrophically while reducing sulfate as a terminal electron acceptor. Optimal growth occurred at pH 7.0 and 28°C with fumarate as electron donor and carbon source. Based on its genome sequence, the G+C content is 40.72mol %. Phylogenomic and phylogenetic analyses indicate that strain BW-1T belongs to the Desulfobacteraceae family within the Deltaproteobacteria class. Based on average amino acid identity, strain BW-1T can be considered as a novel species of a new genus, for which the name Desulfamplus magnetovallimortis is proposed. The type strain of D. magnetovallimortis is BW-1T (JCM 18010T-DSM 103535T).


Asunto(s)
Deltaproteobacteria/clasificación , Deltaproteobacteria/metabolismo , Óxido Ferrosoférrico/metabolismo , Hierro/metabolismo , Sulfuros/metabolismo , Técnicas de Tipificación Bacteriana , Composición de Base/genética , California , ADN Bacteriano/genética , Deltaproteobacteria/genética , Deltaproteobacteria/aislamiento & purificación , Fumaratos/metabolismo , Genoma Bacteriano/genética , Magnetosomas/fisiología , Análisis de Secuencia de ADN
17.
Environ Sci Pollut Res Int ; 24(1): 66-72, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27838908

RESUMEN

The use of biosensors as sensitive and rapid alert systems is a promising perspective to monitor accidental or intentional environmental pollution, but their implementation in the field is limited by the lack of adapted inline water monitoring devices. We describe here the design and initial qualification of an analyzer prototype able to accommodate three types of biosensors based on entirely different methodologies (immunological, whole-cell, and bacteriophage biosensors), but whose responses rely on the emission of light. We developed a custom light detector and a reaction chamber compatible with the specificities of the three systems and resulting in statutory detection limits. The water analyzer prototype resulting from the COMBITOX project can be situated at level 4 on the Technology Readiness Level (TRL) scale and this technical advance paves the way to the use of biosensors on-site.


Asunto(s)
Bacterias/aislamiento & purificación , Bacteriófagos/aislamiento & purificación , Técnicas Biosensibles/métodos , Agua/química , Monitoreo del Ambiente , Análisis de los Alimentos , Luz , Microbiología del Agua , Calidad del Agua
18.
Environ Sci Pollut Res Int ; 24(1): 52-65, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27234828

RESUMEN

Whole-cell biosensors based on reporter genes allow detection of toxic metals in water with high selectivity and sensitivity under laboratory conditions; nevertheless, their transfer to a commercial inline water analyzer requires specific adaptation and optimization to field conditions as well as economical considerations. We focused here on both the influence of the bacterial host and the choice of the reporter gene by following the responses of global toxicity biosensors based on constitutive bacterial promoters as well as arsenite biosensors based on the arsenite-inducible Pars promoter. We observed important variations of the bioluminescence emission levels in five different Escherichia coli strains harboring two different lux-based biosensors, suggesting that the best host strain has to be empirically selected for each new biosensor under construction. We also investigated the bioluminescence reporter gene system transferred into Deinococcus deserti, an environmental, desiccation- and radiation-tolerant bacterium that would reduce the manufacturing costs of bacterial biosensors for commercial water analyzers and open the field of biodetection in radioactive environments. We thus successfully obtained a cell survival biosensor and a metal biosensor able to detect a concentration as low as 100 nM of arsenite in D. deserti. We demonstrated that the arsenite biosensor resisted desiccation and remained functional after 7 days stored in air-dried D. deserti cells. We also report here the use of a new near-infrared (NIR) fluorescent reporter candidate, a bacteriophytochrome from the magnetotactic bacterium Magnetospirillum magneticum AMB-1, which showed a NIR fluorescent signal that remained optimal despite increasing sample turbidity, while in similar conditions, a drastic loss of the lux-based biosensors signal was observed.


Asunto(s)
Arsenitos/química , Técnicas Biosensibles , Deinococcus/metabolismo , Escherichia coli/metabolismo , Luciferasas de la Bacteria/metabolismo , Arsenitos/metabolismo , Deinococcus/genética , Monitoreo del Ambiente/métodos , Escherichia coli/genética , Regulación Bacteriana de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Genes Reporteros , Luciferasas de la Bacteria/genética , Metales Pesados/toxicidad , Regiones Promotoras Genéticas , Agua/química , Contaminantes Químicos del Agua
19.
Biochim Biophys Acta ; 1555(1-3): 54-9, 2002 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-12206891

RESUMEN

It is no surprise that the catalytic activity of electron-transport enzymes may be optimised at certain electrochemical potentials in ways that are analogous to observations of pH-rate optima. This property is observed clearly in experiments in which an enzyme is adsorbed on an electrode surface which can supply or receive electrons rapidly and in a highly controlled manner. In such a way, the rate of catalysis can be measured accurately as a function of the potential (driving force) that is applied. In this paper, we draw attention to a few examples in which this property has been observed in enzymes that are associated with membrane-bound respiratory chains, and we discuss its possible origins and implications for in vivo regulation.


Asunto(s)
Proteínas Hierro-Azufre , Oxidorreductasas/química , Animales , Catálisis , Bovinos , Electroquímica , Transporte de Electrón , Escherichia coli , Concentración de Iones de Hidrógeno , Mitocondrias/enzimología , Nitrato-Reductasa , Nitrato Reductasas , Oxidación-Reducción , Succinato Deshidrogenasa/química
20.
Adv Healthc Mater ; 4(7): 1076-83, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25676134

RESUMEN

The fast development of sensitive molecular diagnostic tools is currently paving the way for a personalized medicine. A new class of ultrasensitive magnetic resonance imaging (MRI) T2-contrast agents based on magnetosomes, magnetite nanocrystals biomineralized by magnetotactic bacteria, is proposed here. The contrast agents can be injected into the blood circulation and detected in the picomolar range. Purified magnetosomes are water-dispersible and stable within physiological conditions and exhibit at 17.2 T a transverse relaxivity r2 four times higher than commercial ferumoxide. The subsequent gain in sensitivity by T2(*) -weighted imaging at 17.2 T of the mouse brain vasculature is evidenced in vivo after tail vein injection of magnetosomes representing a low dose of iron (20 µmoliron kg(-1)), whereas no such phenomenon with the same dose of ferumoxide is observed. Preclinical studies of human pathologies in animal models will benefit from the combination of high magnetic field MRI with sensitive, low dose, easy-to-produce biocompatible contrast agents derived from bacterial magnetosomes.


Asunto(s)
Encéfalo/patología , Óxido Ferrosoférrico/química , Magnetosomas/química , Nanoestructuras/química , Animales , Medios de Contraste/química , Dextranos/química , Imagen por Resonancia Magnética/métodos , Magnetismo/métodos , Nanopartículas de Magnetita/química , Magnetosomas/metabolismo , Magnetospirillum/metabolismo , Ratones , Imagen Molecular/métodos , Nanopartículas/química
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