RESUMEN
Purple non-sulfur bacteria (PNSB) are a diverse group of bacteria studied for various possible applications. They are commonly surveyed in bioenergy research as they produce biohydrogen, a candidate for clean alternative energy. This study aimed to assess the biohydrogen production ability and genetically characterize a high biohydrogen-producing PNSB (MAY2) isolated from Los Baños, Laguna, Philippines via whole genome sequencing (WGS). MAY2, when grown in mixed volatile fatty acids, produced biogas with 38% hydrogen. WGS results revealed that the isolate is positively classified under the genus Rhodobacter johrii. Also, 82 genetic hallmarks for biohydrogen production were found in the isolated genome which are involved in the production of key enzymes and proteins relevant to the photofermentative and hydrogen regulation pathways. Its nitrogenase gene cluster is stringently regulated by two genes, nifA and rofN, whose function and expression are easily affected by several environmental factors.
Asunto(s)
Proteínas Bacterianas , Genoma Bacteriano , Hidrógeno , Rhodobacter , Hidrógeno/metabolismo , Rhodobacter/genética , Rhodobacter/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Secuenciación Completa del Genoma/métodos , Familia de Multigenes , Biocombustibles , Filogenia , Nitrogenasa/genética , Nitrogenasa/metabolismoRESUMEN
Rhodobacter species are promising beneficial microbes that can improve growth performance, immunity and antioxidant capability in aquatic crustaceans. Yet the safety of Rhodobacter azotoformans for potential application in Chinese mitten crab Eriocheir sinensis is still unclear. In the present study, R. azotoformans SY5, a potential probiotic additive that can significantly improve the growth performance, immunity, antioxidant capability, and disease resistance in E. sinensis, was evaluated for safety through whole genome sequencing, antibiotic resistance, toxic metabolites, virulence, and crab tolerance assays. The results indicated that R. azotoformans SY5 only harboured the acyl carrier protein-encoding gene (acpP) that was universal in probiotic bacteria with the function of bacterial fatty acid biosynthesis, exhibited high susceptibility to aminoglycosides, penicillins, polymyxins, polyphosphates, quinolones, and tetracyclines antimicrobials, and possessed inability to produce hemolysin, hydrogen sulphide, nitrite, ammonia, and phenylpyruvate. In addition, R. azotoformans SY5 showed no pathogenicity for E. sinensis with the seven-day acute intraperitoneal LD50 value of above 6.0 × 109 cfu/ml and 30-day chronic oral LD50 of above 6.0 × 109 cfu/g diet. To our knowledge, this is the first report on the safety of R. azotoformans for potential application in Chinese mitten crabs.
Asunto(s)
Antioxidantes , Probióticos , Antioxidantes/metabolismo , Inmunidad Innata , Rhodobacter/metabolismo , Bacterias/metabolismoRESUMEN
Ischemic reperfusion injury (IRI) of the kidneys is a direct sequela of surgical procedures associated with the interruption of blood supply. The pathophysiology of IRI is complicated, and several inflammatories, apoptosis, and oxidative stress pathways are implicated. Among the major receptors directly involved in renal IRI are the toll-like receptors (TLRs), specifically TLR2 and TLR4. In this study, we investigated the effects of Lipopolysaccharide from Rhodobacter Sphaeroides (TLR2 and TLR4 antagonist, LPS-RS) and the ultrapure form (pure TLR4 antagonist, ULPS-RS) on the histopathological changes and TLRs expression in an animal model of bilateral renal IRI. Forty-eight adult male rats were allocated into six groups (N=8) as follows: sham group (negative control without IRI), control group (rats underwent bilateral renal ischemia for 30 minutes and 2 hours of reperfusion), vehicle group (IRI+ vehicle), LPS-RS group (IRI+ 0.5 mg/kg of LPS-RS), ULPS-RS group (IRI+ 0.1 mg/kg of ULPS-RS), ULPS-RSH group (IRI+ 0.2 mg/kg of ULPS-RS). Significant improvement in the histopathological damages induced by renal IRI was found in the ULPS-RS treated groups at both doses compared with the control group. The protective effect of ULPS-RS was associated with significantly reduced TLR4 expression without affecting TLR2. Regarding LPS-RS, the tested dose adversely affected the renal tissues as manifested by the histopathological findings, although it similarly affected TLRs expression as ULPS-RS. Our results demonstrated that ULPS-RS was renoprotective while LPS-RS had no protective effect against the tissue damages induced by renal IRI.
Asunto(s)
Lipopolisacáridos , Daño por Reperfusión , Animales , Riñón/irrigación sanguínea , Lipopolisacáridos/farmacología , Lipopolisacáridos/uso terapéutico , Masculino , Modelos Animales , Ratas , Daño por Reperfusión/complicaciones , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/metabolismo , Rhodobacter/metabolismo , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/metabolismo , Receptor Toll-Like 4/uso terapéutico , Receptores Toll-Like/uso terapéuticoRESUMEN
The effect of exogenous glycine (a precursor for the biosynthesis of bacteriochlorophyll) on the cell growth and photopigment accumulation was investigated in phototrophic growing Rhodobacter azotoformans 134K20. The growth rate and the biomass of strain 134K20 were significantly inhibited by glycine addition when ammonium sulfate or glutamate were used as nitrogen sources and acetate or succinate as carbon sources. A characteristic absorption maximum at approximately 423 nm was present in the absorption spectra of glutamate cultures while it was absent by the addition of high-concentration glycine of 15 mM. The component account for the 423 nm peak was eventually identified as magnesium protoporphyrin IX monomethyl ester, a precursor of bacteriochlorophyll a (BChl a). Comparative analysis of pigment composition revealed that the amount of BChl a precursors was significantly decreased by the addition of 15-mM glycine while the BChl a accumulation was increased. Moreover, glycine changed the carotenoid compositions and stimulated the accumulation of spheroidene. The A850 /A875 in the growth-inhibited cultures was increased, indicating an increased level of the light-harvesting complex 2 compared to the reaction center. The exogenous glycine possibly played an important regulation role in photosynthesis of purple bacteria.
Asunto(s)
Glicina/farmacología , Pigmentos Biológicos/biosíntesis , Rhodobacter/crecimiento & desarrollo , Rhodobacter/metabolismo , Proteínas Bacterianas/metabolismo , Bacterioclorofilas/biosíntesis , Bacterioclorofilas/química , Biomasa , Carotenoides/química , Carotenoides/metabolismo , Fotosíntesis/efectos de los fármacos , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Pigmentos Biológicos/química , Protoporfirinas/metabolismo , Rhodobacter/efectos de los fármacosRESUMEN
Photosynthetic light harvesting can occur with a remarkable near-unity quantum efficiency. The B800-850 complex, also known as light-harvesting complex 2 (LH2), is the primary light-harvesting complex in purple bacteria and has been extensively studied as a model system. The bacteriochlorophylls of the B800-850 complex are organized into two concentric rings, known as the B800 and B850 rings. However, depending on the species and growth conditions, the number of constituent subunits, the pigment geometry, and the absorption energies vary. While the dynamics of some B800-850 variants have been exhaustively characterized, others have not been measured. Furthermore, a direct and simultaneous comparison of how both structural and spectral differences between variants affect these dynamics has not been performed. In this work, we utilize ultrafast transient absorption measurements to compare the B800 to B850 energy-transfer rates in the B800-850 complex as a function of the number of subunits, geometry, and absorption energies. The nonameric B800-850 complex from Rhodobacter (Rb.) sphaeroides is 40% faster than the octameric B800-850 complex from Rhodospirillum (Rs.) molischianum, consistent with structure-based predictions. In contrast, the blue-shifted B800-820 complex from Rs. molischianum is only 20% faster than the B800-850 complex from Rs. molischianum despite an increase in the spectral overlap between the rings that would be expected to produce a larger increase in the energy-transfer rate. These measurements support current models that contain dark, higher-lying excitonic states to bridge the energy gap between rings, thereby maintaining similar energy-transfer dynamics. Overall, these results demonstrate that energy-transfer dynamics in the B800-850 complex are robust to the spectral and structural variations between species used to optimize energy capture and flow in purple bacteria.
Asunto(s)
Complejos de Proteína Captadores de Luz/metabolismo , Rhodobacter/metabolismo , Rhodospirillum/metabolismo , Cristalografía por Rayos X , Transferencia de Energía , Complejos de Proteína Captadores de Luz/química , Modelos Moleculares , Conformación ProteicaRESUMEN
The current research focuses on anaerobic respiration of arsenic and other toxic metals by purple nonsulfur bacteria (PNSB). Among the optimization assays performed were carbon utilization, cross metal resistance, and metal respiration, along with a comparison of each assay in photoheterotrophic and chemoheterotrophic growth. The bacteria were identified by the classification of 16S ribosomal RNA gene sequences. Rhodobacter sp. PI3 proved to be more versatile in carbon source utilization (acetate, lactate, citrate, and oxalate), whereas Rhodopseudomonas palustris PI5 proved to be more versatile in metal resistance (arsenate, arsenite, cobalt, lead, selenium, and nickel). Both the strains were found to be positive for photofermentative hydrogen production along with arsenic respiration. This study reveals that anaerobic conditions are more appropriate for better efficiency of PNSB. Our study demonstrates that R. palustris PI5 and Rhodobacter sp. PI3 can be promising candidates for the biohydrogen production along with metal detoxification using heavy metal-polluted effluents as a substrate.
Asunto(s)
Arsénico/metabolismo , Hidrógeno/metabolismo , Metales/metabolismo , Rhodobacter/metabolismo , Rhodopseudomonas/metabolismo , Anaerobiosis , Biodegradación Ambiental , Carbono/metabolismo , ADN Bacteriano/genética , Procesos Heterotróficos , Filogenia , Pigmentos Biológicos/metabolismo , ARN Ribosómico 16S/genética , Rhodobacter/clasificación , Rhodobacter/genética , Rhodopseudomonas/clasificación , Rhodopseudomonas/genética , Análisis de Secuencia de ADN , Microbiología del AguaRESUMEN
Τhe morphology, physiology and immunology, of solid tumors exhibit spatial heterogeneity which complicates our understanding of cancer progression and therapy response. Understanding spatial heterogeneity necessitates high resolution in vivo imaging of anatomical and pathophysiological tumor information. We introduce Rhodobacter as bacterial reporter for multispectral optoacoustic (photoacoustic) tomography (MSOT). We show that endogenous bacteriochlorophyll a in Rhodobacter gives rise to strong optoacoustic signals >800 nm away from interfering endogenous absorbers. Importantly, our results suggest that changes in the spectral signature of Rhodobacter which depend on macrophage activity inside the tumor can be used to reveal heterogeneity of the tumor microenvironment. Employing non-invasive high resolution MSOT in longitudinal studies we show spatiotemporal changes of Rhodobacter spectral profiles in mice bearing 4T1 and CT26.WT tumor models. Accessibility of Rhodobacter to genetic modification and thus to sensory and therapeutic functions suggests potential for a theranostic platform organism.
Asunto(s)
Técnicas Biosensibles/métodos , Macrófagos/inmunología , Neoplasias/diagnóstico por imagen , Técnicas Fotoacústicas/métodos , Rhodobacter/química , Nanomedicina Teranóstica/métodos , Animales , Bacterioclorofila A/química , Bacterioclorofila A/metabolismo , Línea Celular Tumoral/trasplante , Modelos Animales de Enfermedad , Humanos , Estudios Longitudinales , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Neoplasias/inmunología , Rhodobacter/metabolismo , Tomografía Computarizada por Rayos X/métodos , Microambiente Tumoral/inmunologíaRESUMEN
Fe-organic matter (Fe-OM) complexes are abundant in the environment and, due to their mobility, reactivity, and bioavailability, play a significant role in the biogeochemical Fe cycle. In photic zones of aquatic environments, Fe-OM complexes can potentially be reduced and oxidized, and thus cycled, by light-dependent processes, including abiotic photoreduction of Fe(III)-OM complexes and microbial oxidation of Fe(II)-OM complexes, by anoxygenic phototrophic bacteria. This could lead to a cryptic iron cycle in which continuous oxidation and rereduction of Fe could result in a low and steady-state Fe(II) concentration despite rapid Fe turnover. However, the coupling of these processes has never been demonstrated experimentally. In this study, we grew a model anoxygenic phototrophic Fe(II) oxidizer, Rhodobacter ferrooxidans SW2, with either citrate, Fe(II)-citrate, or Fe(III)-citrate. We found that strain SW2 was capable of reoxidizing Fe(II)-citrate produced by photochemical reduction of Fe(III)-citrate, which kept the dissolved Fe(II)-citrate concentration at low (<10 µM) and stable concentrations, with a concomitant increase in cell numbers. Cell suspension incubations with strain SW2 showed that it can also oxidize Fe(II)-EDTA, Fe(II)-humic acid, and Fe(II)-fulvic acid complexes. This work demonstrates the potential for active cryptic Fe cycling in the photic zone of anoxic aquatic environments, despite low measurable Fe(II) concentrations which are controlled by the rate of microbial Fe(II) oxidation and the identity of the Fe-OM complexes.IMPORTANCE Iron cycling, including reduction of Fe(III) and oxidation of Fe(II), involves the formation, transformation, and dissolution of minerals and dissolved iron-organic matter compounds. It has been shown previously that Fe can be cycled so rapidly that no measurable changes in Fe(II) and Fe(III) concentrations occur, leading to a so-called cryptic cycle. Cryptic Fe cycles have been shown to be driven either abiotically by a combination of photochemical reduction of Fe(III)-OM complexes and reoxidation of Fe(II) by O2, or microbially by a combination of Fe(III)-reducing and Fe(II)-oxidizing bacteria. Our study demonstrates a new type of light-driven cryptic Fe cycle that is relevant for the photic zone of aquatic habitats involving abiotic photochemical reduction of Fe(III)-OM complexes and microbial phototrophic Fe(II) oxidation. This new type of cryptic Fe cycle has important implications for biogeochemical cycling of iron, carbon, nutrients, and heavy metals and can also influence the composition and activity of microbial communities.
Asunto(s)
Bacterias/metabolismo , Compuestos Férricos/metabolismo , Compuestos Ferrosos/metabolismo , Hierro/metabolismo , Procesos Fototróficos/fisiología , Anaerobiosis/fisiología , Carbono/metabolismo , Minerales/metabolismo , Oxidación-Reducción , Rhodobacter/metabolismo , Microbiología del SueloRESUMEN
An ovoid to rod-shaped, phototrophic, purple non-sulfur bacterium was isolated from a sediment sample of a hot spring in Tibet, China. Cells of strain YIM 73036T were Gram-stain negative, non-motile and multiplied by binary fission. Strain YIM 73036T grew optimally at pH 7.0-7.5 at 37-45 °C. Growth occurred in 0.5-3.5% (w/v) NaCl. Vitamins were not required for growth. The presence of photosynthesis genes pufL and pufM were shown and photosynthesis pigments were formed. Bacteriochlorophyll α, the bacteriopheophytin and carotenoids were present as photosynthetic pigments. Internal cytoplasmic membranes were of the lamellar type. The organism YIM 73036T was able to grow chemo-organoheterophically, chemo-lithoautotrophically and photo-organoheterotrophically but photo-lithoautotrophic and fermentative growth were not demonstrated. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain YIM 73036T is closely related to Rhodobacter blasticus ATCC 33485T (96.65% sequence similarity) and clustered with species of the genus Rhodobacter of the family Rhodobacteraceae. Whole-genome sequence analyses based on the average nucleotide BLAST identity (ANI < 82%) indicated that this isolate belongs to a novel species. The genomic DNA G+C content of organism YIM 73036T was determined to be 66.0 mol%. Strain YIM 73036T contained Q-10 as the predominant ubiquinone and C18:1ω7c, C18:1ω7c 11-methyl and C18:0 as the major fatty acids. The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and unidentified phospholipid. Differential phenotypic and chemotaxonomic properties, together with the phylogenetic distinctiveness, demonstrated that strain YIM 73036T is distinguishable from other species of the genus Rhodobacter. On the basis of the data presented, strain YIM 73036T is considered to represent a novel species of the genus Rhodobacter, for which the name Rhodobacter thermarum sp. nov. [type strain YIM 73036T (= KCTC 52712T = CCTCC AB 2016298T)] is proposed.
Asunto(s)
Sedimentos Geológicos/microbiología , Manantiales de Aguas Termales/microbiología , Rhodobacter/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Ácidos Grasos/metabolismo , Sedimentos Geológicos/química , Manantiales de Aguas Termales/química , Filogenia , ARN Ribosómico 16S/genética , Rhodobacter/clasificación , Rhodobacter/genética , Rhodobacter/metabolismo , Cloruro de Sodio/análisis , Cloruro de Sodio/metabolismo , TibetRESUMEN
An exopolysaccharide (EPS) producing bacterial strain was isolated from the surface of marine macroalgae (Padina sp.). Based on polyphasic taxonomy, the strain CDR-SL 7Cii was assigned to the genus Rhodobacter and found to be the closest relative of the species Rhodobacter johrii. The strain was able to produce 6.2â¯g/l of EPS upon fermentation using R2A medium enriched with 2.5% glucose. FT-IR analysis revealed the presence of hydroxyl, carboxyl and diacyl-ester functional groups in the purified EPS. Further Chromatographic study revealed that R. johrii synthesized a high molecular weight anionic exopolysaccharide composed of glucose, glucuronic acid, rhamnose and galactose in a molar ratio of 3:1.5:0.25:0.25. The 1D and 2D NMR spectroscopy (COSY/HSQC) analysis revealed the presence of 1,6 linked-α-d-Glcp, 1,4 linked-ß-d-Glcp, 1,3 linked-ß-d-GlcA, 1,3 linked-ß-d-Galp, 1,6 linked-ß-d-Galf and 3-α-l-Rhmp residues. Moreover, the purified EPS has shown stability towards elevated temperature and also acted as a bio-emulsifier to create a high pH and temperature stable emulsion of hydrocarbon/water indicating its industrial potential. This is the first report of EPS production by a strain of Rhodobacter johrii.
Asunto(s)
Organismos Acuáticos , Calor , Polisacáridos Bacterianos , Rhodobacter , Organismos Acuáticos/química , Organismos Acuáticos/metabolismo , Conformación de Carbohidratos , Polisacáridos Bacterianos/biosíntesis , Polisacáridos Bacterianos/química , Polisacáridos Bacterianos/aislamiento & purificación , Rhodobacter/química , Rhodobacter/metabolismoRESUMEN
The effects of ultraviolet (UV) irradiation (up to 0.6 J/cm2) and heating (65 °C, 20 min) on the absorption spectra and electron transfer in dehydrated film samples of photosynthetic reaction centers (RCs) from purple bacterium Rhodobacter (Rb.) sphaeroides, as well as in hybrid structures consisting of RCs and quantum dots (QDs), have been studied. The samples were placed in organic matrices containing the stabilizers of protein structure-polyvinyl alcohol (PVA) and trehalose. UV irradiation led to partially irreversible oxidation of some RCs, as well as to transformation of some fraction of the bacteriochlorophyll (BChl) molecules into bacteriopheophytin (BPheo) molecules. In addition, UV irradiation causes degradation of some BChl molecules that is accompanied by formation of 3-acetyl-chlorophyll a molecules. Finally, UV irradiation destroys the RCs carotenoid molecules. The incorporation of RCs into organic matrices reduced pheophytinization. Trehalose was especially efficient in reducing the damage to the carotenoid and BChl molecules caused by UV irradiation. Hybrid films containing RC + QD were more stable to pheophytinization upon UV irradiation. However, the presence of QDs in films did not affect the processes of carotenoid destruction. The efficiency of the electronic excitation energy transfer from QD to P865 also did not change under UV irradiation. Heating led to dramatic destruction of the RCs structure and bacteriochlorins acquired the properties of unbound molecules. Trehalose provided strong protection against destruction of the RCs and hybrid (RC + QD) complexes.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Puntos Cuánticos , Rayos Ultravioleta , Calefacción , Rhodobacter/metabolismo , Trehalosa/metabolismoRESUMEN
The Salar de Huasco is an evaporitic basin located in the Chilean Altiplano, which presents extreme environmental conditions for life, i.e. high altitude (3800 m.a.s.l.), negative water balance, a wide salinity range, high daily temperature changes and the occurrence of the highest registered solar radiation on the planet (> 1200 W m-2). This ecosystem is considered as a natural laboratory to understand different adaptations of microorganisms to extreme conditions. Rhodobacter, an anoxygenic aerobic phototrophic bacterial genus, represents one of the most abundant groups reported based on taxonomic diversity surveys in this ecosystem. The bacterial mat isolate Rhodobacter sp. strain Rb3 was used to study adaptation mechanisms to stress-inducing factors potentially explaining its success in a polyextreme ecosystem. We found that the Rhodobacter sp. Rb3 genome was characterized by a high abundance of genes involved in stress tolerance and adaptation strategies, among which DNA repair and oxidative stress were the most conspicuous. Moreover, many other molecular mechanisms associated with oxidative stress, photooxidation and antioxidants; DNA repair and protection; motility, chemotaxis and biofilm synthesis; osmotic stress, metal, metalloid and toxic anions resistance; antimicrobial resistance and multidrug pumps; sporulation; cold shock and heat shock stress; mobile genetic elements and toxin-antitoxin system were detected and identified as potential survival mechanism features in Rhodobacter sp. Rb3. In total, these results reveal a wide set of strategies used by the isolate to adapt and thrive under environmental stress conditions as a model of polyextreme environmental resistome.
Asunto(s)
Adaptación Fisiológica/genética , Ecosistema , Ambientes Extremos , Rhodobacter/fisiología , Microbiología del Agua , Altitud , Chile , Biología Computacional , ADN Bacteriano/genética , Genes Bacterianos , Modelos Biológicos , ARN Ribosómico 16S/genética , ARN de Transferencia/genética , Rhodobacter/genética , Rhodobacter/metabolismo , Especificidad de la EspecieRESUMEN
Biohydrogen production via fermentative routes offers considerable advantages in waste recycling and sustainable energy production. This can be realized by single-stage dark or photofermentative processes, or by a two-stage integrated process; the latter offering the higher production yields due to complete conversion of sugar substrates into H2 and CO2. However, problems arising from the integration of these two processes limit its scale-up and implementation. Hence, high efficiency one-step fermentative biohydrogen production processes from sugar-rich wastes are preferable. In this study, different strains of purple non-sulfur bacteria were investigated for their biohydrogen production capacity on pure sucrose and sugar beet molasses, and the feasibility of single-stage photofermentative biohydrogen production was evaluated. A single-stage photofermentation process was carried out using four different strains of purple non-sulfur bacteria (Rhodobacter capsulatus DSM 1710, R. capsulatus YO3, Rhodobacter sphaeroides O.U.001, and Rhodopseudomonas palustris DSM 127) on different initial sucrose concentrations. The highest hydrogen yield obtained was 10.5 mol H2/mol of sucrose and the maximum hydrogen productivity was 0.78 mmol/L h by Rp. palustris on 5 mM sucrose. A hydrogen yield of 19 mol H2/mol sucrose, which represents 79% of theoretical yield, and a maximum hydrogen productivity of 0.55 mmol/L h were obtained by Rp. palustris from sugar beet molasses. The yield was comparable to those values obtained in two-stage processes. The present study demonstrates that single-stage photofermentation using purple non-sulfur bacteria on sucrose-based wastes is promising.
Asunto(s)
Beta vulgaris/microbiología , Fermentación , Hidrógeno/metabolismo , Melaza , Rhodobacter/metabolismo , Concentración de Iones de Hidrógeno , Fotoquímica , Rhodobacter/crecimiento & desarrollo , Especificidad de la Especie , Sacarosa/metabolismoRESUMEN
Mixed microbial cultures (MMC) and waste/surplus substrates, as hardwood spent sulfite liquor, are being used to decrease polyhydroxyalkanoates' (PHA) production costs. The process involves two or three steps, being the selection step a crucial one. For the industrial implementation of this strategy, reactor stability in terms of both performance and microbial community presence has to be considered. A long-term operation of a sequencing batch reactor under feast/famine conditions was performed along with microbial community identification/quantification using FISH and DGGE. The community was found to be extremely dynamic, dominated by Alphaproteobacteria, with Paracoccus and Rhodobacter present, both PHA-storing microorganisms. 16S rRNA gene clone library further revealed that side populations' non-PHA accumulators were able to strive (Agrobacterium, Flavobacteria, and Brachymonas). Nevertheless, reactor performance in terms of PHA storage was stable during operation time. The monitoring of the MMC population evolution provided information on the relation between community structure and process operation.
Asunto(s)
Medios de Cultivo/química , Microbiología Industrial , Polihidroxialcanoatos/biosíntesis , Agrobacterium/aislamiento & purificación , Agrobacterium/metabolismo , Alphaproteobacteria/aislamiento & purificación , Alphaproteobacteria/metabolismo , Reactores Biológicos/microbiología , Clonación Molecular , Comamonadaceae/aislamiento & purificación , Comamonadaceae/metabolismo , ADN Bacteriano/aislamiento & purificación , Flavobacterium/aislamiento & purificación , Flavobacterium/metabolismo , Biblioteca de Genes , Hibridación Fluorescente in Situ , Paracoccus/aislamiento & purificación , Paracoccus/metabolismo , Filogenia , Polihidroxialcanoatos/análisis , Rhodobacter/aislamiento & purificación , Rhodobacter/metabolismoRESUMEN
Energy-coupling factor (ECF) transporters are a large family of ATP-binding cassette transporters recently identified in microorganisms. Responsible for micronutrient uptake from the environment, ECF transporters are modular transporters composed of a membrane substrate-binding component EcfS and an ECF module consisting of an integral membrane scaffold component EcfT and two cytoplasmic ATP binding/hydrolysis components EcfA/A'. ECF transporters are classified into groups I and II. Currently, the molecular understanding of group-I ECF transporters is very limited, partly due to a lack of transporter complex structural information. Here, we present structures and structure-based analyses of the group-I cobalt ECF transporter CbiMNQO, whose constituting subunits CbiM/CbiN, CbiQ, and CbiO correspond to the EcfS, EcfT, and EcfA components of group-II ECF transporters, respectively. Through reconstitution of different CbiMNQO subunits and determination of related ATPase and transporter activities, the substrate-binding subunit CbiM was found to stimulate CbiQO's basal ATPase activity. The structure of CbiMQO complex was determined in its inward-open conformation and that of CbiO in ß, γ-methyleneadenosine 5'-triphosphate-bound closed conformation. Structure-based analyses revealed interactions between different components, substrate-gating function of the L1 loop of CbiM, and conformational changes of CbiO induced by ATP binding and product release within the CbiMNQO transporter complex. These findings enabled us to propose a working model of the CbiMNQO transporter, in which the transport process requires the rotation or toppling of both CbiQ and CbiM, and CbiN might function in coupling conformational changes between CbiQ and CbiM.
Asunto(s)
Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Cobalto/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Rhodobacter/metabolismo , Adenosina Trifosfato/metabolismo , Proteínas de Transporte de Membrana/química , Modelos Biológicos , Modelos Moleculares , Unión Proteica , Estructura Secundaria de Proteína , Subunidades de Proteína/metabolismoRESUMEN
Two strains of photosynthetic bacteria, Rhodobacter blasticus and Rhodobacter capsulatus, were used in this work to investigate the feasibility of using photosynthetic bacteria for the treatment of anaerobically digested swine wastewater. The effects of crucial factors which influence the pollutants removal efficiency were also examined. Results showed that anaerobically digested swine wastewater could be treated effectively by photosynthetic bacteria. The treatment efficiency was significantly higher by the mixed photosynthetic bacteria than that by any unitary bacterium. The optimal treatment condition by mixed bacteria was inoculation of 10.0%(v/v) of the two bacteria by 1:1, initial pH of 7.0 and initial chemical oxygen demand of 4800mgL-1. Under these conditions, the removal rate of chemical oxygen demand was 83.3%, which was 19.3% higher than when using Rhodobacter blasticus or 10.6% higher than when using Rhodobacter capsulatus separately. This mixed photosynthetic bacteria achieved high chemical oxygen demand removal and cell yields.
Asunto(s)
Rhodobacter/metabolismo , Aguas Residuales/microbiología , Purificación del Agua/métodos , Anaerobiosis , Animales , Análisis de la Demanda Biológica de Oxígeno , Biomasa , Estudios de Factibilidad , Concentración de Iones de Hidrógeno , Fotosíntesis , Rhodobacter capsulatus/metabolismo , PorcinosRESUMEN
The exponential rise in energy demand vis-à-vis depletion of mineral oil resources has accelerated recovery of bioenergy from organic waste. In this study, a laboratory-scale anaerobic (An)/aerobic (Ar) system comprising of expanded granular sludge bed (EGSB) reactor coupled to an aerobic sequential batch reactor (SBR) was constructed to treat beet sugar industrial wastewater (BSIW) of chemical oxygen demand (COD) 1665 mg L-1 while harnessing methane gas. The EGSB reactor generated methane at the rate of 235 mL/g COD added, with considerably higher than previously reported methane content of 86 %. Meanwhile, contaminants were successfully reduced in the combined An/Ar system, realizing a removal rate of more than 71.4, 97.3, 97.7, and 99.3 % of organic matter as total phosphorus, total nitrogen, biological oxygen demand (BOD), and soluble COD, respectively. Microbial community analysis showed that the bacterial genus Clostridium sp. and archaeal genus Methanosaeta sp. dominated the EGSB reactor, while Rhodobacter sp. dominance was observed in the SBR. The obtained experimental results indicate that the integration of expanded granular sludge bed and sequential batch reactor in treating BSIW obtained competitively outstanding performance.
Asunto(s)
Beta vulgaris , Reactores Biológicos , Residuos Industriales , Eliminación de Residuos Líquidos/métodos , Aerobiosis , Anaerobiosis , Análisis de la Demanda Biológica de Oxígeno , Reactores Biológicos/microbiología , Clostridium/metabolismo , Metano/biosíntesis , Methanosarcinales/metabolismo , Nitrógeno/metabolismo , Fósforo/metabolismo , Rhodobacter/metabolismo , Aguas Residuales , Contaminantes Químicos del Agua/metabolismoRESUMEN
Chlorophylls are light-capturing units found in photosynthetic proteins. We study here the ground and excited state properties of monomeric, dimeric, and tetrameric models of the special chlorophyll/bacteriochlorophyll (Chl/BChl) pigment (P) centers P700 and P680/P870 of type I and type II photosystems, respectively. In the excited state calculations, we study the performance of the algebraic diagrammatic construction through second-order (ADC(2)) method in combination with the reduced virtual space (RVS) approach and the recently developed Laplace-transformed scaled-opposite-spin (LT-SOS) algorithm, which allows us, for the first time, to address multimeric effects at correlated ab initio levels using large basis sets. At the LT-SOS-RVS-ADC(2)/def2-TZVP level, we obtain vertical excitation energies (VEEs) of 2.00-2.07 and 1.52-1.62 eV for the P680/P700 and the P870 pigment models, respectively, which agree well with the experimental absorption maxima of 1.82, 1.77, and 1.43 eV for P680, P700, and P870, respectively. In the P680/P870 models, we find that the photoexcitation leads to a π â π* transition in which the exciton is delocalized between the adjacent Chl/BChl molecules of the central pair, whereas the exciton is localized to a single chlorophyll molecule in the P700 model. Consistent with experiments, the calculated excitonic splittings between the central pairs of P680, P700, and P870 models are 80, 200, and 400 cm(-1), respectively. The calculations show that the electron affinity of the radical cation of the P680 model is 0.4 V larger than for the P870 model and 0.2 V larger than for P700. The chromophore stacking interaction is found to strongly influence the electron localization properties of the light-absorbing pigments, which may help to elucidate mechanistic details of the charge separation process in type I and type II photosystems.
Asunto(s)
Clorofila/química , Complejo de Proteína del Fotosistema I/química , Complejo de Proteína del Fotosistema II/química , Transporte de Electrón , Luz , Fotosíntesis , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Estructura Terciaria de Proteína , Teoría Cuántica , Rhodobacter/metabolismo , TermodinámicaRESUMEN
Translocator protein 18 kDa (TSPO) was previously known as the peripheral benzodiazepine receptor (PBR) in eukaryotes, where it is mainly localized to the mitochondrial outer membrane. Considerable evidence indicates that it plays regulatory roles in steroidogenesis and apoptosis and is involved in various human diseases, such as metastatic cancer, Alzheimer's and Parkinson's disease, inflammation, and anxiety disorders. Ligands of TSPO are widely used as diagnostic tools and treatment options, despite there being no clear understanding of the function of TSPO. An ortholog in the photosynthetic bacterium Rhodobacter was independently discovered as the tryptophan-rich sensory protein (TspO) and found to play a role in the response to changes in oxygen and light conditions that regulate photosynthesis and respiration. As part of this highly conserved protein family found in all three kingdoms, the rat TSPO is able to rescue the knockout phenotype in Rhodobacter, indicating functional as well as structural conservation. Recently, a major breakthrough in the field was achieved: the determination of atomic-resolution structures of TSPO from different species by several independent groups. This now allows us to reexamine the function of TSPO with a molecular perspective. In this review, we focus on recently determined structures of TSPO and their implications for potential functions of this ubiquitous multifaceted protein. We suggest that TSPO is an ancient bacterial receptor/stress sensor that has developed additional interactions, partners, and roles in its mitochondrial outer membrane environment in eukaryotes.