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1.
ISME J ; 15(11): 3384-3398, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34054125

RESUMO

Photoferrotrophy allows anoxygenic phototrophs to use reduced iron as an electron donor for primary productivity. Recent work shows that freshwater photoferrotrophs can use electrons from solid-phase conductive substances via phototrophic extracellular electron uptake (pEEU), and the two processes share the underlying electron uptake mechanism. However, the ability of marine phototrophs to perform photoferrotrophy and pEEU, and the contribution of these processes to primary productivity is largely unknown. To fill this knowledge gap, we isolated 15 new strains of the marine anoxygenic phototroph Rhodovulum sulfidophilum on electron donors such as acetate and thiosulfate. We observed that all of the R. sulfidophilum strains isolated can perform photoferrotrophy. We chose strain AB26 as a representative strain to study further, and find that it can also perform pEEU from poised electrodes. We show that during pEEU, AB26 transfers electrons to the photosynthetic electron transport chain. Furthermore, systems biology-guided mutant analysis shows that R. sulfidophilum AB26 uses a previously unknown diheme cytochrome c protein, which we call EeuP, for pEEU but not photoferrotrophy. Homologs of EeuP occur in a range of widely distributed marine microbes. Overall, these results suggest that photoferrotrophy and pEEU contribute to the biogeochemical cycling of iron and carbon in marine ecosystems.


Assuntos
Elétrons , Rhodovulum , Transporte Biológico , Ecossistema , Rhodovulum/genética
2.
Commun Biol ; 3(1): 357, 2020 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-32641733

RESUMO

Photosynthetic microorganisms such as cyanobacteria, purple bacteria and microalgae have attracted great interest as promising platforms for economical and sustainable production of bioenergy, biochemicals, and biopolymers. Here, we demonstrate heterotrophic production of spider dragline silk proteins, major ampullate spidroins (MaSp), in a marine photosynthetic purple bacterium, Rhodovulum sulfidophilum, under both photoheterotrophic and photoautotrophic growth conditions. Spider silk is a biodegradable and biocompatible material with remarkable mechanical properties. R. sulfidophilum grow by utilizing abundant and renewable nonfood bioresources such as seawater, sunlight, and gaseous CO2 and N2, thus making this photosynthetic microbial cell factory a promising green and sustainable production platform for proteins and biopolymers, including spider silks.


Assuntos
Reatores Biológicos , Fibroínas/biossíntese , Rhodovulum/metabolismo , Animais , Reatores Biológicos/microbiologia , Fibroínas/genética , Fibroínas/isolamento & purificação , Fibroínas/ultraestrutura , Microrganismos Geneticamente Modificados/genética , Microrganismos Geneticamente Modificados/metabolismo , Microscopia Eletrônica de Varredura , Fotossíntese , Rhodovulum/genética , Aranhas
3.
Microbiologyopen ; 9(1): e00953, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31638342

RESUMO

Marine purple photosynthetic bacteria are ideal organisms for the production of useful materials at reduced costs and contributing to a sustainable society because they can utilize sunlight, seawater, and components of air, including carbon dioxide and nitrogen gases, for their growth. However, conjugation is the only applicable method for the transformation of marine purple photosynthetic bacteria so far. Here, we examined a calcium chloride-mediated method for the transformation of marine purple photosynthetic bacteria. Plasmid DNAs containing the kanamycin resistance gene were successfully transferred into chemically competent cells of two strains of marine purple photosynthetic bacteria (Rhodovulum sulfidophilum and Roseospira marina). Heat shock treatment increased the transformation efficiency in R. sulfidophilum, whereas the addition of cell-penetrating peptide did not improve it. We also found that prolonged incubation in agar plates containing kanamycin led to spontaneous mutation of the 16S rRNA, resulting in kanamycin resistance in R. marina. Thus, we developed an efficient and facile transformation method using chemically competent cells of marine purple photosynthetic bacteria with calcium chloride.


Assuntos
Técnicas de Transferência de Genes , Resistência a Canamicina/genética , Rhodospirillaceae/genética , Rhodovulum/genética , Transformação Bacteriana/genética , Cloreto de Cálcio/química , DNA Bacteriano/química , DNA Bacteriano/genética , Resposta ao Choque Térmico/fisiologia , Plasmídeos/genética , Água do Mar/microbiologia , Microbiologia da Água
4.
Microbiol Res ; 215: 130-140, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30172299

RESUMO

Biofixation of CO2 is being extensively investigated to solve the global warming problem. Purple non-sulfur bacteria are fast growers that consume CO2 and produce beneficial biomass. Better the growth at higher CO2 levels, more efficient are the strains for biofixation. Nine among fifty strains that were analyzed at elevated CO2 levels responded with better growth. Considering its enhanced growth at high CO2 and metabolic versatility, Rhodovulum viride strain JA756 was chosen to make further studies. Strain JA756 tolerates up to 50% (v/v) CO2 with its optimum between 20-40% (v/v), yielding a biomass of 3.4 g. L-1. The pattern of specific enzyme activity of carbonic anhydrase corresponded well with that of its growth. To gain insights into the genomic composition and genes related to carbonic anhydrases and CO2 fixation, draft genome sequencing of JA756 was carried out which revealed the presence of two non-homologous genes encoding for ß and γ carbonic anhydrases, both of which are assumed to be implicated in maintaining intracellular inorganic carbon concentration at equilibrium. Most of the genes involved in the Calvin pathway, reductive tricarboxylic acid pathway, 3-hydroxypropionate bicycle and C4 pathways were found in the draft genome. While the experimental determinations of active roles of two of these pathways are still underway, the expression of key genes of Calvin and C4 pathway suggest their functional role in the organism. Owing to its metabolic versatility, JA756 can be advantageous for biological CO2 assimilation facilities located by the coastline, inland and also at wide ranges of CO2 concentrations.


Assuntos
Ciclo do Carbono/fisiologia , Dióxido de Carbono/metabolismo , Anidrases Carbônicas/metabolismo , Rhodovulum/enzimologia , Rhodovulum/metabolismo , Processos Autotróficos/genética , Processos Autotróficos/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biomassa , Carbono/metabolismo , Ciclo do Carbono/genética , Dióxido de Carbono/administração & dosagem , Dióxido de Carbono/farmacologia , Anidrases Carbônicas/genética , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genes Bacterianos/genética , Aquecimento Global , Cinética , Ácido Láctico/análogos & derivados , Ácido Láctico/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/fisiologia , Fotossíntese/genética , Rhodovulum/genética , Rhodovulum/crescimento & desenvolvimento , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
5.
J Gen Appl Microbiol ; 64(3): 103-107, 2018 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-29526925

RESUMO

The marine bacterium Rhodovulum sulfidophilum is a nonsulfur phototrophic bacterium, which is known to produce extracellular nucleic acids in soluble form in culture medium. In the present paper, constructing the response regulator ctrA-deficient mutant of R. sulfidophilum, we found that this mutation causes a significant decrease in the extracellular DNA production. However, by the introduction of a plasmid containing the wild type ctrA gene into the mutant, the amount of extracellular DNA produced was recovered. This is the first and clear evidence that the extracellular DNA production is actively controlled by the CtrA in R. sulfidophilum.


Assuntos
Proteínas de Bactérias/genética , DNA Bacteriano/biossíntese , Espaço Extracelular/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Rhodovulum/genética , Rhodovulum/metabolismo , Organismos Aquáticos/genética , Organismos Aquáticos/metabolismo , DNA Bacteriano/metabolismo , Teste de Complementação Genética , Mutagênese Insercional , Plasmídeos/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
6.
FEMS Microbiol Lett ; 365(3)2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29228187

RESUMO

Extracellular nucleic acids of high molecular weight are detected ubiquitously in seawater. Recent studies have indicated that these nucleic acids are, at least in part, derived from active production by some bacteria. The marine bacterium Rhodovulum sulfidophilum is one of those bacteria. Rhodovulumsulfidophilum is a non-sulfur phototrophic marine bacterium that is known to form structured communities of cells called flocs, and to produce extracellular nucleic acids in culture media. Recently, it has been revealed that this bacterium produces gene transfer agent-like particles and that this particle production may be related to the extracellular nucleic acid production mechanism. This review provides a summary of recent physiological and genetic studies of these phenomena and also introduces a new method for extracellular production of artificial and biologically functional RNAs using this bacterium. In addition, artificial RNA production using Escherichia coli, which is related to this topic, will also be described.


Assuntos
Espaço Extracelular/metabolismo , Microbiologia Industrial/métodos , Ácidos Nucleicos/metabolismo , RNA Bacteriano/biossíntese , Rhodovulum/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Espaço Extracelular/química , Floculação , Ácidos Nucleicos/biossíntese , Ácidos Nucleicos/genética , RNA/biossíntese , RNA/genética , RNA Bacteriano/genética , Rhodovulum/genética , Rhodovulum/crescimento & desenvolvimento , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
7.
Bioengineered ; 8(5): 670-677, 2017 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-28282262

RESUMO

RNA interference-based technologies have emerged as an attractive and effective therapeutic option with potential application in diverse human diseases. These tools rely on the development of efficient strategies to obtain homogeneous non-coding RNA samples with adequate integrity and purity, thus avoiding non-targeted gene-silencing and related side-effects that impair their application onto pre-clinical practice. These RNAs have been preferentially obtained by in vitro transcription using DNA templates or via chemical synthesis. As an alternative to overcome the limitations presented by these methods, in vivo recombinant production of RNA biomolecules has become the focus in RNA synthesis research. Therefore, using pre-miR-29b as a model, here it is evaluated the time-course profile of Escherichia coli and Rhodovolum sulfidophilum microfactories to produce this microRNA. As the presence of major host contaminants arising from the biosynthesis process may have important implications in the subsequent downstream processing, it is also evaluated the production of genomic DNA and host total proteins. Considering the rapidly growing interest on these innovative biopharmaceuticals, novel, more cost-effective, simple and easily scaled-up technologies are highly desirable. As microRNA recombinant expression fulfills those requirements, it may take the leading edge in the methodologies currently available to obtain microRNAs for clinical or structural studies.


Assuntos
Reatores Biológicos/microbiologia , Escherichia coli/genética , MicroRNAs/biossíntese , MicroRNAs/genética , Recombinação Genética/genética , Rhodovulum/genética , Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Humanos , Rhodovulum/metabolismo
8.
Int J Syst Evol Microbiol ; 66(9): 3367-3371, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27260502

RESUMO

A reddish-brown-pigmented, phototrophic bacterium, designated strain JA877T, was isolated from a brown algae mat sample collected from Jalandhar beach, Gujarat, India. On the basis of the 16S rRNA gene sequence, strain JA877T belongs to the class Alphaproteobacteria and is closely related to the type strains Rhodovulum viride JA756T (99.0 %), Rhodovulum sulfidophilum Hansen W4T (98.9 %), Rhodovulumvisakhapatnamense JA181T (98.8 %),Rhodovulum kholense JA297T (97.5 %) and Rhodovulum salis JA746T (97.0). However, strain JA877T showed only 20-45 % relatedness with its phylogenetic neighbours and had a ∆Tm between 5.8 and 7.0 °C. The major respiratory quinone was ubiquinone-10 (Q10), and the polar lipid profile was composed of the major components phosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid, two unidentified sulfolipids and five unidentified lipids. The major fatty acids were C18 : 1ω5c, C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C18 : 0. The DNA G+C content was 64.5 mol%. On the basis of 16S rRNA gene sequence analysis, physiological data, and chemotaxonomic and molecular differences, strain JA877T is significantly different from other species of the genus Rhodovulum and represents a novel species, for which the name Rhodovulum algae sp. nov. is proposed. The type strain is JA877T (=LMG 29228T= KCTC 42963T).


Assuntos
Phaeophyceae/microbiologia , Filogenia , Rhodovulum/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Índia , Lipídeos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Rhodovulum/genética , Rhodovulum/isolamento & purificação , Análise de Sequência de DNA , Ubiquinona/química
9.
Appl Microbiol Biotechnol ; 100(8): 3723-34, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26860940

RESUMO

The present study reports the successful production of human pre-miR-29b both intra- and extracellularly in the marine phototrophic bacterium Rhodovulum sulfidophilum using recombinant RNA technology. In a first stage, the optimal transformation conditions (0.025 µg of plasmid DNA, with a heat-shock of 2 min at 35 °C) were established, in order to transfer the pre-miR-29b encoding plasmid to R. sulfidophilum host. Furthermore, the extracellular recovery of this RNA product from the culture medium was greatly improved, achieving quantities that are compatible with the majority of applications, namely for in vitro or in vivo studies. Using this system, the extracellular human pre-miR-29b concentration was approximately 182 µg/L, after 40 h of bacterial growth, and the total intracellular pre-miR-29b was of about 358 µg/L, at 32 h. At the end of the fermentation, it was verified that almost 87 % of cells were viable, indicating that cell lysis is minimized and that the extracellular medium is not highly contaminated with the host intracellular ribonucleases (RNases) and endotoxins, which is a critical parameter to guarantee the microRNA (miRNA) integrity. These findings demonstrate that pre-miRNAs can be produced by recombinant RNA technology, offering novel clues for the production of natural pre-miRNA agents for functional studies and RNA interference (RNAi)-based therapeutics.


Assuntos
Expressão Gênica , MicroRNAs/biossíntese , Rhodovulum/metabolismo , Meios de Cultura/metabolismo , Humanos , MicroRNAs/genética , Plasmídeos/genética , Plasmídeos/metabolismo , Rhodovulum/genética
10.
Int J Syst Evol Microbiol ; 66(1): 165-171, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26475698

RESUMO

A yellowish brown, phototrophic, purple non-sulfur bacterium, strain JA924T, was isolated in pure culture from a brackish water sample collected from an estuary. Single cells were oval to rod-shaped, non-motile and Gram-stain-negative and had a vesicular architecture of intracellular photosynthetic membranes. Bacteriochlorophyll-a and carotenoids of the spheroidene series were present as photosynthetic pigments. Photolithoautotrophy, chemo-organoheterotrophy and photo-organoheterotrophy were the growth modes observed. Strain JA924T had complex growth requirements. Strain JA924T was mesophilic and moderately halophilic. The DNA G+C content was 64 mol% (HPLC). The major cellular fatty acids were C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C18 : 0. The major quinone was ubiquinone-10 (Q-10). Phosphatidylglycerol, phosphatidylethanolamine, sulfolipid and an aminolipid were the main polar lipids of strain JA924T. EzTaxon-e blast searches based on the 16S rRNA gene sequence of JA924T revealed highest similarity with Rhodovulum mangrovi AK41T (98.19 %) and other members of the genus Rhodovulum ( < 95.71 %). Strain JA924T was further identified to be distantly related to Rhodovulum mangrovi AK41T ( < 29 % based on DNA-DNA hybridization and ΔTm (>5 °C). Phenotypic, chemotaxonomic and molecular differences indicate that strain JA924T represents a novel species of the genus Rhodovulum, for which the name Rhodovulum aestuarii sp. nov. is proposed. The type strain is JA924T ( = LMG 29031T = KCTC 15485T).


Assuntos
Filogenia , Rhodovulum/classificação , Águas Salinas , Técnicas de Tipagem Bacteriana , Bacterioclorofila A , Composição de Bases , Carotenoides/química , DNA Bacteriano/genética , Estuários , Ácidos Graxos/química , Índia , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Rhodovulum/genética , Rhodovulum/isolamento & purificação , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/química
11.
Int J Syst Evol Microbiol ; 64(Pt 9): 3168-3173, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24972612

RESUMO

A novel Gram-staining-negative, purple non-sulfur bacterium, strain AK41(T), was isolated from a sediment sample collected from Coringa mangrove forest, Andhra Pradesh, India. A red-brownish-coloured culture was obtained on modified Pfennig medium after enrichment with 2 % NaCl and 0.3 % pyruvate under 2000 lx illumination. Individual cells were ovoid-rod-shaped and non-motile. Bacteriochlorophyll a and carotenoids of the spheroidene series were present as photosynthetic pigments. Strain AK41(T) was halophilic and grew photoheterotrophically with a number of organic compounds as carbon sources and electron donors. It was unable to grow photoautotrophically. It did not utilize sulfide or thiosulfate as electron donors. The fatty acids were found to be dominated by C16 : 0 and C18 : 1ω7c. Strain AK41(T) contained phosphatidylglycerol, phosphatidylethanolamine, an unknown aminolipid and four unknown lipids as polar lipids. Q-10 was the predominant respiratory quinone. The DNA G+C content of strain AK41(T) was 68.9 mol%. 16S rRNA gene sequence analysis indicated that strain AK41(T) was a member of the genus Rhodovulum and was closely related to Rhodovulum sulfidophilum, with 96.0 % similarity to the type strain; the 16S rRNA gene sequence similarity to the type strains of other species of the genus Rhodovulum was 93.9-95.8 %. Phylogenetic analyses indicated that strain AK41(T) clustered with the type strains of Rhodovulum marinum, Rdv. kholense, Rdv. sulfidophilum and Rdv. visakhapatnamense with sequence similarity of 95.9-96.2 %. Based on data from the current study, strain AK41(T) is proposed to represent a novel species of the genus Rhodovulum, for which the name Rhodovulum mangrovi sp. nov. is proposed. The type strain of Rhodovulum mangrovi is AK41(T) ( = MTCC 11825(T) = JCM 19220(T)).


Assuntos
Sedimentos Geológicos/microbiologia , Filogenia , Rhodovulum/classificação , Áreas Alagadas , Avicennia/microbiologia , Técnicas de Tipagem Bacteriana , Bacterioclorofila A/química , Composição de Bases , Carotenoides/química , DNA Bacteriano/genética , Ácidos Graxos/química , Índia , Dados de Sequência Molecular , Pigmentação , RNA Ribossômico 16S/genética , Rhodovulum/genética , Rhodovulum/isolamento & purificação , Análise de Sequência de DNA , Ubiquinona/química
12.
Artigo em Inglês | MEDLINE | ID: mdl-24508674

RESUMO

Recently, differential expression of microRNAs, in patients with Alzheimer's disease (AD) suggests that they might have key regulatory roles in this neurodegenerative disease. Taking into account this fact, several studies demonstrated that the miR-29 is significantly decreased in AD patients, also displaying abnormally high levels of ß-site APP-cleaving enzyme 1. Thus, RNA biochemical or structural studies often require a RNA sample that is chemically pure and biologically active. The present work describes a new affinity chromatography method using an arginine support to specifically purify pre-miR-29 from other Rhodovulum sulfidophilum small RNA species. Nevertheless, in order to achieve higher efficiency and selectivity, it is essential to characterize the behavior of pre-miR-29 binding/elution. Thus, three different strategies based on increased sodium chloride (280-500mM), arginine (25mM) or decreased ammonium sulfate (2-0.1M) stepwise gradients are described to purify pre-miR-29. In this way, it was proved that well-defined binding/elution conditions are crucial to enhance the purification performance. As a matter of fact, by employing elution strategies using sodium chloride or arginine, an improvement in the final pre-miR-29 yields (96.5 and 56.7%, respectively) was obtained. Moreover, the quality control analysis revealed high integrity in pre-miR-29 preparations as well as high purity (90 and 98%, respectively), demonstrated by the scarce detection of proteins. This improved method takes advantage of its simplicity, significant cost reduction, due to the elimination of some complex operations, and speed for large-scale purification of pre-miRNAs suitable for biochemical and structural studies.


Assuntos
Arginina/química , Cromatografia de Afinidade/métodos , MicroRNAs/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , MicroRNAs/análise , MicroRNAs/química , MicroRNAs/genética , Concentração Osmolar , Rhodovulum/química , Rhodovulum/genética , Temperatura
13.
Bioresour Technol ; 154: 254-9, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24412477

RESUMO

Most of the photosynthetic bacterial strains exhibit optimum hydrogen production at neutral initial pH, and lower initial pH resulted in a sharp decrease in hydrogen yield. Thus, screening of acid-tolerant hydrogen-producing photosynthetic bacteria is very important. To obtain acid tolerant mutants, a Tn7-based transposon was randomly inserted into the genomic DNA of Rhodovulum sulfidophilum P5. An acid tolerant mutant strain TH-102 exhibited increased hydrogen production in acidic environment (pH 4.5-6.5) and at higher temperatures (35 and 37°C) than the wild-type strain. At pH 5.5 and 35°C, the mutant strain TH-102 continuously produced hydrogen. The hydrogen yield and average rate were 2.16 ± 0.10 mol/mol acetate and 10.06 ± 0.47 mL/Lh, which was about 17.32 and 15.37-fold higher than that of the wild-type strain, respectively. This acid- and temperature-tolerant mutant strain TH-102 could be used in a cost-effective hydrogen production process employing both dark fermentative and photosynthetic bacteria.


Assuntos
Ácidos/farmacologia , Adaptação Fisiológica/efeitos dos fármacos , Elementos de DNA Transponíveis/genética , Hidrogênio/metabolismo , Mutação/genética , Fotobiologia/métodos , Rhodovulum/genética , Técnicas de Cultura Celular por Lotes , Mutagênese/efeitos dos fármacos , Mutagênese/genética , Reação em Cadeia da Polimerase , Rhodovulum/efeitos dos fármacos , Temperatura
14.
Int J Syst Evol Microbiol ; 64(Pt 3): 957-962, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24425825

RESUMO

Two strains (JA746(T) and JA756(T)) having yellowish brown-to-green pigment were isolated from a solar saltern and a pink pond, respectively. While both strains were non-motile and shared the presence of bacteriochlorophyll-a, major cellular fatty acids (C18 : 1ω7c, C16 : 0, C18 : 0), quinone (Q-10), polar lipids and hopanoids, they differed from each other in their carotenoid composition. The G+C content of genomic DNA of strains JA746(T) and 756(T) was 62.4 and 63.3 mol%, respectively. The 16S rRNA gene-based EzTaxon-e blast search analysis of strains JA746(T) and 756(T) indicated highest sequence similarity with members of the genus Rhodovulum in the family Rhodobacteraceae of the class Alphaproteobacteria. Strain JA746(T) has high sequence similarities with Rhodovulum visakhapatnamense JA181(T) (97.3 %), Rhodovulum steppense A-20s(T) (97.3 %), Rhodovulum phaeolacus JA580(T) (97 %), Rhodovulum strictum MB-G2(T) (97 %) and other members of the genus Rhodovulum (<97 %). Strain JA756(T) has high sequence similarities with Rhodovulum visakhapatnamense JA181(T) (99.8 %), Rhodovulum sulfidophilum Hansen W4(T) (99.1 %), Rhodovulum kholense JA297(T) (97.9 %) and other members of the genus Rhodovulum (<97 %). The sequence similarity between strains JA746(T) and JA756(T) was 97.5 %. However, these strains are not closely related to each other or to their phylogenetic neighbours since the DNA-DNA reassociation values were less than 56 %. The genomic information was also supported by phenotypic and chemotaxonomic results, leading us to classify strains JA746(T) ( = NBRC 108898(T) = KCTC 15180(T)) and JA756(T) ( = NBRC 109122(T) = KCTC 15223(T)) as the type strains of two novel species of the genus Rhodovulum, for which the names Rhodovulum salis sp. nov. and Rhodovulum viride sp. nov. are proposed, respectively.


Assuntos
Filogenia , Lagoas/microbiologia , Rhodovulum/classificação , Microbiologia da Água , Bacterioclorofila A/química , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Índia , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Rhodovulum/genética , Rhodovulum/isolamento & purificação , Água do Mar , Análise de Sequência de DNA , Cloreto de Sódio , Ubiquinona/química
15.
J Gen Appl Microbiol ; 60(6): 222-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25742972

RESUMO

Previously, we proposed a new method for production of RNA aptamers using the marine bacterium Rhodovulum sulfidophilum. A streptavidin RNA aptamer (an RNA which binds to streptavidin) was extracellularly produced by this bacterium containing engineered plasmid. The aptamer had full biological function. As a next step we attempted to produce another functional RNA, short hairpin RNAs (shRNAs) using this bacterial system. We have designed two types of shRNAs targeted to the luciferase gene. Here we report that shRNAs are successfully produced extracellularly by this system. Even if the shRNA has a long stem-loop structure which is thought to interfere with transcription in bacterial cells, the yield of the shRNA is almost the same as that of the streptavidin RNA aptamer. During the course of these experiments, we also found a new type of RNA processing for the double-stranded region of the shRNA.


Assuntos
Organismos Aquáticos/metabolismo , Engenharia Metabólica , RNA Interferente Pequeno/biossíntese , Rhodovulum/metabolismo , Organismos Aquáticos/genética , Luciferases/genética , Luciferases/metabolismo , Plasmídeos , RNA Interferente Pequeno/genética , Rhodovulum/genética
16.
Bioresour Technol ; 142: 18-25, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23732918

RESUMO

In this study, transposon mutagenesis technology was utilized to enhance the hydrogen production capability of a wild marine photosynthetic bacterium Rhodovulum sulfidophilum P5. A mutant strain TH-253 that exhibited high hydrogen yield and weaker light absorption ability was screened. Under strong light conditions, the mutant produced more hydrogen than that of the WT. Under optimum light intensity (120 µmol photons/m(2)s), the mutant achieved its highest hydrogen yield (1,436 ± 44 mL H2/L, about 3.21 ± 0.10 mol H2/mol acetate), which was 40.37% higher that of the WT. In continuous operation mode, the hydrogen yield (3.59 ± 0.11 mol H2/mol acetate) and average hydrogen production rate (16.91 ± 0.46 mL H2/Lh) of the mutant were 43.40% and 45.07% higher than those of the WT, respectively. The mutant strain TH-253 may be used as an appropriate starting strain for future photosynthesis-based large scale hydrogen production.


Assuntos
Hidrogênio/metabolismo , Mutação , Rhodovulum/metabolismo , Sequência de Bases , Primers do DNA , Elementos de DNA Transponíveis , Genes Bacterianos , Reação em Cadeia da Polimerase , Rhodovulum/genética , Rhodovulum/crescimento & desenvolvimento
17.
J Biol Chem ; 287(48): 40350-9, 2012 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-23060437

RESUMO

BACKGROUND: SoxAX enzymes initiate microbial oxidation of reduced inorganic sulfur compounds. Their catalytic mechanism is unknown. RESULTS: Cyanide displaces the CysS(-) ligand to the active site heme following reduction by S(2)O(4)(2-) but not Eu(II). CONCLUSION: An active site heme ligand becomes labile on exposure to substrate analogs. SIGNIFICANCE: Elucidation of SoxAX mechanism is necessary to understand a widespread pathway for sulfur compound oxidation. SoxAX enzymes couple disulfide bond formation to the reduction of cytochrome c in the first step of the phylogenetically widespread Sox microbial sulfur oxidation pathway. Rhodovulum sulfidophilum SoxAX contains three hemes. An electrochemical cell compatible with magnetic circular dichroism at near infrared wavelengths has been developed to resolve redox and chemical properties of the SoxAX hemes. In combination with potentiometric titrations monitored by electronic absorbance and EPR, this method defines midpoint potentials (E(m)) at pH 7.0 of approximately +210, -340, and -400 mV for the His/Met, His/Cys(-), and active site His/CysS(-)-ligated heme, respectively. Exposing SoxAX to S(2)O(4)(2-), a substrate analog with E(m) ~-450 mV, but not Eu(II) complexed with diethylene triamine pentaacetic acid (E(m) ~-1140 mV), allows cyanide to displace the cysteine persulfide (CysS(-)) ligand to the active site heme. This provides the first evidence for the dissociation of CysS(-) that has been proposed as a key event in SoxAX catalysis.


Assuntos
Proteínas de Bactérias/química , Grupo dos Citocromos c/química , Heme/metabolismo , Oxirredutases/química , Rhodovulum/enzimologia , Enxofre/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Catálise , Domínio Catalítico , Grupo dos Citocromos c/genética , Grupo dos Citocromos c/metabolismo , Cinética , Oxirredução , Oxirredutases/genética , Oxirredutases/metabolismo , Rhodovulum/química , Rhodovulum/genética
18.
J Bacteriol ; 194(22): 6363, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23105089

RESUMO

We report the 4.8-Mb draft genome of Rhodovulum sp. strain PH10, a phototrophic bacterium belonging to class Alphaproteobacteria, isolated from a soil sample collected from the mangrove forest of Namkhana in India. This genome is the first from the genus Rhodovulum and will lead to a better understanding of the genes/pathways involved in activities like phototrophic growth and nitrogen fixation in this group of bacteria.


Assuntos
Genoma Bacteriano , Rhodovulum/classificação , Rhodovulum/genética , Microbiologia do Solo , Áreas Alagadas , Índia , Dados de Sequência Molecular , Rhodovulum/isolamento & purificação
19.
J Fish Dis ; 35(12): 927-34, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22943666

RESUMO

RNA aptamers are artificial nucleic acids that specifically bind to a wide variety of targets. They are an effective tool for pharmaceutical research and development of antiviral agents. Here, we describe four Hirame rhabdovirus (HIRRV)-RNA aptamers (H1, H2, H3 and H4) that we obtained from an in vitro process called the systematic evolution of ligands by exponential enrichment (SELEX). The HIRRV-RNA aptamers specifically bind to HIRRV. Hirame natural embryo (HINAE) cells treated with virus and the RNA aptamer showed a decrease in appearance of cytopathic effect when compared with control (treated only with virus). Rhodovulum sulfidophilum was transformed with genes for the RNA aptamers, and the aptamers were detected in the culture medium, indicating that they were secreted from the cells. Thus, the recombinant R. sulfidophilum might be a powerful tool for the prevention of HIRRV in aquaculture.


Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Aptâmeros de Nucleotídeos/farmacologia , Novirhabdovirus/crescimento & desenvolvimento , Replicação Viral/efeitos dos fármacos , Animais , Antivirais/farmacologia , Aptâmeros de Nucleotídeos/biossíntese , Linhagem Celular , DNA Recombinante/genética , Linguado , Plasmídeos/genética , Rhodovulum/genética
20.
Int J Syst Evol Microbiol ; 62(Pt 12): 2962-2966, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22286907

RESUMO

Two strains of purple non-sulfur bacteria (A-36s(T) and A-51s) were isolated from brackish steppe soda lakes of southern Siberia. Genetically, the isolates were related most closely to the type strains of Rhodovulum steppense and Rhodovulum strictum, from which they differed at the species level (98.5% 16S rRNA gene sequence similarity, 40-53% DNA-DNA relatedness). Cells of the two strains were ovoid to rod-shaped, 0.4-0.8 µm wide and 1.0-2.5 µm long, and motile by means of a polar flagellum. They contained internal photosynthetic membranes of vesicular type and photosynthetic pigments (bacteriochlorophyll a and carotenoids of the spheroidene series). The strains were obligate haloalkaliphiles, growing over wide ranges of salinity (0.3-10.0% NaCl) and pH (7.5-10.0), with growth optima at 1.0-3.0% NaCl and pH 8.5-9.0. Photoheterotrophic and chemoheterotrophic growth occurred with a number of organic compounds and biotin, p-aminobenzoate, thiamine and niacin as growth factors. No anaerobic respiration on nitrite, nitrate or fumarate and no fermentation were demonstrated. The strains grew photolithoautotrophically and chemolithoautotrophically with sulfide, sulfur and thiosulfate, oxidizing them to sulfate. Sulfide was oxidized via deposition of extracellular elemental sulfur. No growth with H(2) as the electron donor was observed. The major fatty acid was C(18:1) (78%). The major quinone was ubiquinone Q-10. The DNA G+C content of strain A-36s(T) was 65.4 mol% (T(m)). According to genotypic and phenotypic characteristics, the investigated strains were assigned to a novel species of the genus Rhodovulum, for which the name Rhodovulum tesquicola sp. nov. is proposed. The type strain is A-36s(T) ( = VKM B-2491(T) = ATCC BAA-1573(T)), which was isolated from steppe soda lake Sul'fatnoe (Zabaikal'skii Krai, southern Siberia, Russia).


Assuntos
Lagos/microbiologia , Filogenia , Rhodovulum/classificação , Microbiologia da Água , DNA Bacteriano/genética , Ácidos Graxos/análise , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Rhodobacteraceae/classificação , Rhodobacteraceae/genética , Rhodobacteraceae/isolamento & purificação , Rhodovulum/genética , Rhodovulum/isolamento & purificação , Análise de Sequência de DNA , Sibéria , Sulfatos/metabolismo , Ubiquinona/análise
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