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1.
Microbiol Res ; 247: 126733, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33676313

RESUMEN

Since sugarcane is a ratoon crop, genome analysis of plant growth-promoting bacteria that exist in its soil rhizosphere, can provide opportunity to better understand their characteristics and use of such bacteria in turn, may especially improve perennial crop productivity. In the present study, genome of two bacterial strains, one each of B. megaterium (BM89) and B. subtilis (BS87), isolated and reported earlier (Chandra et al., 2018), were sequenced and characterized. Though both strains have demonstrated plant growth promoting properties and enhanced in-vitro plant growth responses, functional annotation and analysis of genes indicated superiority of BS87 as it possessed more plant growth promotion attributable genes over BM89. Apart from some common genes, trehalose metabolism, glycine betaine production, peroxidases, super oxide dismutase, cold shock proteins and phenazine production associated genes were selectively identified in BS87 genome indicating better plant growth performances and survival potential under harsh environmental conditions. Genes for chitinase, d-cysteine desulfhydrase and γ-aminobutyric acid (GABA), as found in BM89, propose its selective utilization in defense and bio-control measures. Concomitant with better settlings' growth, scanning electron micrographs indicated these isolated and characterized bacteria exhibiting healthy colonization within root of sugarcane crop. Kegg pathways' assignment also revealed added pathways namely carbohydrate and amino acid metabolism attached to B. subtilis strain BS87, a preferable candidate for bio-fertilizer and its utilization to promote growth of both plant and ratoon crops of sugarcane usually experiencing harsh environmental conditions.


Asunto(s)
Bacillus megaterium/genética , Bacillus subtilis/genética , Desarrollo de la Planta , Rizosfera , Saccharum/crecimiento & desarrollo , Saccharum/microbiología , Secuenciación Completa del Genoma , Bacillus megaterium/clasificación , Bacillus megaterium/aislamiento & purificación , Bacillus megaterium/fisiología , Bacillus subtilis/clasificación , Bacillus subtilis/aislamiento & purificación , Bacillus subtilis/fisiología , Proteínas y Péptidos de Choque por Frío , Producción de Cultivos , Productos Agrícolas/microbiología , Fertilizantes , Genoma Bacteriano , Filogenia , Suelo , Microbiología del Suelo
2.
Antonie Van Leeuwenhoek ; 113(9): 1263-1278, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32564275

RESUMEN

Plant growth-promoting rhizobacteria are a group of free-living bacteria that colonize plant rhizosphere and benefit plant root growth, thereby increasing host plant to cope with salinity induced stress. The aim of this study was to (1) isolate and characterize auxin-producing bacteria showing a high plant growth-promoting (PGP) potential, and (2) evaluate the PGP effects on the growth of Medicago sativa L under salinity stress (130 mM NaCl). Of thirteen isolates, Bacillus megaterium NRCB001 (NRCB001), B. subtilis subsp. subtilis NRCB002 (NRCB002) and B. subtilis NRCB003 (NRCB003) had the ability to produce auxin, which ranged from 47.53 to 154.38 µg ml-1. The three auxin-producing bacterial strains were shown multiple PGP traits, such as producing siderophore and NH3, showing ACC deaminase activity, solubilize phosphate and potassium. Furthermore, NRCB001, NRCB002, and NRCB003 could survive in LB medium containing 1750 mM NaCl. The three auxin-producing with salinity tolerance strains were selected for further analyses. In greenhouse experiments, when inoculated with NRCB001, NRCB002 and NRCB003, dry weight of alfalfa significantly (P < 0.05) increased by 24.1%, 23.1% and 38.5% respectively, compared with those of non-inoculated control seedlings under normal growth condition. When inoculated with NRCB002 and NRCB003, dry weight of alfalfa significantly (P < 0.05) increased by 96.9 and 71.6% respectively, compared with those of non-inoculated control seedlings under 130 mM NaCl condition. Our results indicated that NRCB002 and NRCB003 having PGP traits are promising candidate strains to develop biofertilizers, especially used under salinity stress conditions.


Asunto(s)
Bacillus megaterium/fisiología , Bacillus subtilis/fisiología , Ácidos Indolacéticos/metabolismo , Medicago sativa/crecimiento & desarrollo , Raíces de Plantas/microbiología , Salinidad , Bacillus megaterium/clasificación , Bacillus megaterium/aislamiento & purificación , Bacillus subtilis/clasificación , Bacillus subtilis/aislamiento & purificación , ADN Bacteriano/genética , Medicago sativa/microbiología , Filogenia , Desarrollo de la Planta , ARN Ribosómico 16S/genética , Rizosfera , Cloruro de Sodio , Microbiología del Suelo
3.
Int J Biol Macromol ; 161: 1181-1188, 2020 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-32561282

RESUMEN

An exopolysaccharide (EPS)-producing bacterial PFY-147 was separated from Hiqiher vineyard soil and determined as Bacillus megaterium by 16S rDNA analysis and morphological. The PFY-147-EPS was purified by gel-filtration chromatography. The average molecular weight (Mw) of purified EPS was 1.878 × 106 Da. The total carbohydrate contents, uronic acid and sulfated group content in PFY-147-EPS were found to be 90.37 ± 1.48%, 7.24 ± 0.36% and 2.39 ± 0.36%, respectively. The PFY-147-EPS was found to be a levan containing a backbone of 2,6-substituted ß-fructoses by high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) and Fourier transform infrared (FT-IR) spectroscopy. The atomic force microscopy (AFM) presented that the levan formed pointed or thorny structural in aqueous solution. Scanning electron microscopy (SEM) of the levan showed compact and nonuniform block sizes morphology. Moreover, the levan exhibited higher thermal stability with a degradation temperature of 265.63 °C in DSC analysis. The levan possessed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, Superoxide anion (O2-) radical, hydroxyl (OH) radical and and2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+) scavenging activities and strong metal ion chelating activity. The WHC and WSI of levan were 231.29 ± 3.76% and 97.34 ± 1.72%, respectively. These might have implications in food and medicine, as well as in pharmaceutical industries.


Asunto(s)
Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Bacillus megaterium/química , Fructanos/química , Fructanos/aislamiento & purificación , Fructanos/farmacología , Adsorción , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Fraccionamiento Químico , Espectroscopía de Resonancia Magnética , Metales/química , Microscopía de Fuerza Atómica , Peso Molecular , Monosacáridos/química , Filogenia , Solubilidad , Espectroscopía Infrarroja por Transformada de Fourier
4.
Curr Microbiol ; 76(10): 1215-1224, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31254008

RESUMEN

Nejayote is an alkaline wastewater generated during the nixtamalization process. Nejayote contains high-value compounds such as ferulic acid (FA), which is widely employed as a substrate for the biotechnological production of flavors and aromas. In the present study, the isolation, identification, and characterization of a native strain of Bacillus megaterium were performed, and its capacity to produce 4-vinylguaiacol (4VG) from ferulic acid was evaluated by employing growing cell and resting cell systems. Growing cells of native B. megaterium biotransformed 6 mM crude FA in nejayote into 2.1 mM 4VG, reaching a productivity of 0.21 mM h-1 4VG, while nejayote enriched with FA at 10, 15, and 25 mM resulted in the formation of 2.4, 3.8, and 6.2 mM 4VG and productivities of 0.24, 0.38, and 0.51 mM h-1 4VG, respectively. In the resting cell system, from 6 and 25 mM pure FA, 3.5 mM 4VG was produced (0.18 mM h-1 4VG), while at 10 and 15 mM FA, 4.6 and 5.1 mM 4VG (average of 0.24 mM h-1 4VG) were obtained, respectively. The native B. megaterium strain, isolated from nejayote, showed great biotechnological potential to produce 4VG from crude FA contained in this wastewater, in which other Bacillus species, such as B. licheniformis and B. cereus, were unable to grow and biotransform FA into 4VG.


Asunto(s)
Bacillus megaterium/clasificación , Bacillus megaterium/metabolismo , Ácidos Cumáricos/metabolismo , Aguas Residuales/microbiología , Zea mays , Bacillus megaterium/genética , Bacillus megaterium/crecimiento & desarrollo , Biomasa , Biotransformación , Ácidos Cumáricos/química , Guayacol/análogos & derivados , Guayacol/metabolismo , Filogenia , Aguas Residuales/química
5.
FEMS Microbiol Lett ; 366(22)2019 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-31913456

RESUMEN

In the present study, the taxonomic position of Bacillus aryabhattai and Bacillus megaterium was evaluated using morphological, biochemical, phylogenomic and genome analysis. The morphological and biochemical of these two species were almost similar with few exceptions. The major fatty acids in B. megaterium DSM 32T and B. aryabhattai 21047T were anteiso-C15:0 and iso-C15:0. In the phylogenomic tree, both species clade together and shared high 16S rRNA gene sequence similarity (99.6%). The average nucleotide identity values between Bacillus aryabhattai and Bacillus megaterium were above the threshold values for bacterial species delineation. Based upon morphological, biochemical, chemotaxonomic and comparative genome analysis, we propose to reclassify Bacillus aryabhattai Shivaji et al. 2009 as a later heterotypic synonym of Bacillus megaterium de Bary 1884 (Approved Lists 1980).


Asunto(s)
Bacillus megaterium/clasificación , Bacillus megaterium/genética , Bacillus/clasificación , Bacillus/genética , Bacillus/citología , Bacillus/fisiología , Bacillus megaterium/citología , Bacillus megaterium/fisiología , Análisis por Conglomerados , Citosol/química , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Genómica , Técnicas de Tipificación Micológica , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico
6.
J Gen Appl Microbiol ; 65(3): 137-144, 2019 Jul 19.
Artículo en Inglés | MEDLINE | ID: mdl-30568045

RESUMEN

An extracellular L-asparaginase was isolated and purified from Bacillus megaterium MG1 to apparent homogeneity. The purification procedure involved a combination of ammonium sulfate precipitation, ion-exchange chromatography, and gel filtration techniques, resulting in a purification factor of 31.52 fold with a specific activity of 215 U mg-1. The molecular mass of the purified enzyme was approximately 47 kDa on SDS-PAGE and 185 kDa on native PAGE gel as well as in gel filtration column chromatography, revealing that the enzyme was a homotetramer. The Km and Vmax values of the purified enzyme were calculated to be 2.0 ⅹ 10-4 M and 1.198 mM s-1. Maximum enzyme activity was observed over a wide range of temperature and pH values with an optimum temperature of 37°C and pH 8.5. SDS and metal ions such as Fe2+, Cu2+, Mg2+, Co2+, Mn2+, and Ca2+ decreased the enzyme activity remarkably, whereas the addition of Na+ and K+ led to an increase in activity. The insensitivity of the protein in the presence of EDTA suggested that the enzyme might not essentially be a metalloprotein. Its marked stability and activity in organic solvents and reducing agents suggest that this asparaginase is highly suitable as a biotechnological tool with industrial applications.


Asunto(s)
Asparaginasa/aislamiento & purificación , Asparaginasa/metabolismo , Bacillus megaterium/enzimología , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Microbiología del Agua , Asparaginasa/química , Asparaginasa/genética , Asparagina/metabolismo , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Bacillus megaterium/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Activación Enzimática , Estabilidad de Enzimas , Espacio Extracelular/metabolismo , Bosques , Concentración de Iones de Hidrógeno , India , Cinética , Peso Molecular , Filogenia , Especificidad por Sustrato , Temperatura
7.
PLoS One ; 13(4): e0196166, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29694403

RESUMEN

The intense use of herbicides for weed control in agriculture causes selection pressure on soil microbiota and water ecosystems, possibly resulting in changes to microbial processes, such as biogeochemical cycles. These xenobiotics may increase the production of reactive oxygen species and consequently affect the survival of microorganisms, which need to develop strategies to adapt to these conditions and maintain their ecological functionality. This study analyzed the adaptive responses of bacterial isolates belonging to the same species, originating from two different environments (water and soil), and subjected to selection pressure by herbicides. The effects of herbicide Callisto and its active ingredient, mesotrione, induced different adaptation strategies on the cellular, enzymatic, and structural systems of two Bacillus megaterium isolates obtained from these environments. The lipid saturation patterns observed may have affected membrane permeability in response to this herbicide. Moreover, this may have led to different levels of responses involving superoxide dismutase and catalase activities, and enzyme polymorphisms. Due to these response systems, the strain isolated from water exhibited higher growth rates than did the soil strain, in evaluations made in oligotrophic culture media, which would be more like that found in semi-pristine aquatic environments. The influence of the intracellular oxidizing environments, which changed the mode of degradation of mesotrione in our experimental model and produced different metabolites, can also be observed in soil and water at sites related to agriculture. Since the different metabolites may present different levels of toxicity, we suggest that this fact should be considered in studies on the fate of agrochemicals in different environments.


Asunto(s)
Bacillus megaterium/crecimiento & desarrollo , Ciclohexanonas/farmacología , Herbicidas/farmacología , Microbiología del Suelo , Microbiología del Agua , Adaptación Fisiológica , Bacillus megaterium/clasificación , Bacillus megaterium/efectos de los fármacos , Bacillus megaterium/genética , Biodegradación Ambiental , Ecosistema , Peroxidación de Lípido/efectos de los fármacos , Viabilidad Microbiana/efectos de los fármacos , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
8.
Benef Microbes ; 7(4): 609-20, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27291405

RESUMEN

The interaction between the enteric microbiota and intestinal cells often involves signal molecules that affect both microbial behaviour and host responses. Examples of such signal molecules are the molecules secreted by bacteria that induce quorum sensing mechanisms in the producing microorganism and signal transduction pathways in the host cells. The pentapeptide competence and sporulation factor (CSF) of Bacillus subtilis is a well characterized quorum sensing factor that controls competence and spore formation in the producing bacterium and induces cytoprotective heat shock proteins in intestinal epithelial cells. We analysed several Bacillus strains isolated from human ileal biopsies of healthy volunteers and observed that some of them were unable to produce CSF but still able to act in a CSF-like fashion on model intestinal epithelial cells. One of those strains belonging to the Bacillus megaterium species secreted at least two factors with effects on intestinal HT29 cells: a peptide smaller than 3 kDa able to induce heat shock protein 27 (hsp27) and p38-MAPK, and a larger molecule able to induce protein kinase B (PKB/Akt) with a pro-proliferative effect.


Asunto(s)
Bacillus megaterium/fisiología , Células Epiteliales/microbiología , Bacillus megaterium/clasificación , Bacillus megaterium/aislamiento & purificación , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/uso terapéutico , Células CACO-2 , Ciclo Celular , Línea Celular Tumoral , Medios de Cultivo , Células Epiteliales/citología , Proteínas de Choque Térmico HSP27/metabolismo , Células HT29 , Humanos , Percepción de Quorum , Proteínas Represoras/metabolismo , Proteínas Represoras/uso terapéutico , Transducción de Señal , Estrés Fisiológico
9.
Biosci Biotechnol Biochem ; 80(7): 1264-73, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26967471

RESUMEN

Light is a ubiquitous environmental factor serving as an energy source and external stimulus. Here, I review the conserved molecular mechanism of light-inducible production of carotenoids in three nonphototrophic bacteria: Streptomyces coelicolor A3(2), Thermus thermophilus HB27, and Bacillus megaterium QM B1551. A MerR family transcriptional regulator, LitR, commonly plays a central role in their light-inducible carotenoid production. Genetic and biochemical studies on LitR proteins revealed a conserved function: LitR in complex with adenosyl B12 (AdoB12) has a light-sensitive DNA-binding activity and thus suppresses the expression of the Crt biosynthesis gene cluster. The in vitro DNA-binding and transcription assays showed that the LitR-AdoB12 complex serves as a repressor allowing transcription initiation by RNA polymerase in response to illumination. The existence of novel light-inducible genes and the unique role of the megaplasmid were revealed by the transcriptomic analysis of T. thermophilus. The findings suggest that LitR is a general regulator responsible for the light-inducible carotenoid production in the phylogenetically divergent nonphototrophic bacteria, and that LitR performs diverse physiological functions in bacteria.


Asunto(s)
Bacillus megaterium/efectos de la radiación , Proteínas Bacterianas/genética , Carotenoides/biosíntesis , Proteínas de Unión al ADN/genética , Regulación Bacteriana de la Expresión Génica , Streptomyces coelicolor/efectos de la radiación , Thermus thermophilus/efectos de la radiación , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Bacillus megaterium/metabolismo , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Proteínas de Unión al ADN/metabolismo , ARN Polimerasas Dirigidas por ADN/genética , ARN Polimerasas Dirigidas por ADN/metabolismo , Luz , Familia de Multigenes , Filogenia , Plásmidos/química , Plásmidos/metabolismo , Regiones Promotoras Genéticas , Streptomyces coelicolor/clasificación , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Thermus thermophilus/clasificación , Thermus thermophilus/genética , Thermus thermophilus/metabolismo , Transcripción Genética , Vitamina B 12/metabolismo
10.
N Biotechnol ; 32(6): 682-9, 2015 Dec 25.
Artículo en Inglés | MEDLINE | ID: mdl-25982267

RESUMEN

Biodegradable polymers are currently viable alternatives to traditional synthetic polymers. For instance, polyhydroxybutyrate (PHB) is intracellularly produced and accumulated by Bacillus species, among others. This study reports several wild-type Bacillus strains with the ability to accumulate PHB using raw glycerol from biodiesel production as the sole carbon source. Out of 15 strains from different sources, B. megaterium B2 was selected as the most promising strain for further statistical optimization of the medium composition. Plackett-Burman and central composite designs were used to establish key variables and optimal culture conditions for PHB production using both 250-mL shake flasks and a 7.5-L bioreactor. Temperature and concentrations of glycerol and Na2HPO4 are the experimental variables with the most significant influence on PHB production by B2. After 14 hours of fermentation in shake flasks with optimized medium, B2 produced 0.43 g/L of PHB with a 34% accumulation in the cells. In contrast, under the same conditions, a maximum PHB concentration of 1.20 g/L in the bioreactor was reached at 11 hours. These values correspond to a 48% and 314% increase in PHB production compared to the initial culture conditions. These results suggest the potential of B2 as a PHB producer using raw glycerol, which is an inexpensive, abundant and readily available carbon source.


Asunto(s)
Bacillus megaterium/metabolismo , Biocombustibles/microbiología , Glicerol/metabolismo , Hidroxibutiratos/metabolismo , Residuos Industriales/prevención & control , Valor Nutritivo/fisiología , Poliésteres/metabolismo , Bacillus megaterium/clasificación , Biodegradación Ambiental , Reactores Biológicos/microbiología , Glicerol/aislamiento & purificación , Hidroxibutiratos/aislamiento & purificación , Fosfatos/metabolismo , Poliésteres/aislamiento & purificación , Temperatura
11.
Int J Biol Macromol ; 76: 254-61, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25697675

RESUMEN

Oceans have significant potential to empower mankind and thus marine organisms are believed to be an enormous source for useful biomolecules. Polyhydroxyalkanoates (PHAs) are biological macromolecules that can be applied in nearly all fields. In the present study, Bacillus megaterium strain JK4h has been exploited for maximum PHB production using novel Dry Sea Mix (DSM) via Central Composite Design (CCD) of Response Surface Methodology (RSM) approach. The isolate was found to be producing 56.77% Cell Dry Weight (CDW) of PHAs within 24h, with optimized combinations of peptone, yeast extract and glucose. The PHB yield had been increased 2.61 fold compared to un-optimized experiments. The obtained PHA/PHB had been chemically characterized through Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FTIR), Thermo Gravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC). The results indicate the successful optimization for maximum production of biological macromolecule and it was found to be highly pure polyhydroxybutyrate (PHB). Thus, DSM can be served as a novel and cost effective medium for PHA production offering the use of marine resources as a "green" sustainable alternative.


Asunto(s)
Bacillus megaterium/metabolismo , Polihidroxialcanoatos/química , Polihidroxialcanoatos/farmacología , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Rastreo Diferencial de Calorimetría , Fraccionamiento Químico , Genes Bacterianos , Resonancia Magnética Nuclear Biomolecular , Filogenia , Reproducibilidad de los Resultados , Espectroscopía Infrarroja por Transformada de Fourier , Termogravimetría
12.
Wei Sheng Wu Xue Bao ; 55(12): 1543-50, 2015 Dec 04.
Artículo en Chino | MEDLINE | ID: mdl-27101696

RESUMEN

OBJECTIVE: The aim of this study was to screen tobacco straw and nicotine degrading microorganism. METHODS: The bacterium was isolated from tobacco field soil using medium containing tobacco straw as the sole carbon and nitrogen source. We identified the bacterium through morphological and physiological characterization combined with the result of 16S rRNA gene sequence and data analysis. We also studied the lignocelluloses degradation and enzyme activities related to the degradation of lignin and cellulose in liquid state fermentation of tobacco stalk. RESULTS: The bacterium was identified as Bacillus megaterium and we had demonstrated that it has a good ability to degrade lignin in tobacco straw when fermented in liquid state. It showed the highest laccase production of 418. 52 U/L while the highest lignin peroxides and manganese peroxides activity was 19. 71 U/L and 64. 71 U/L. On the other hand, we also found that nicotine in tobacco stem was totally degraded 20 d after inoculation. CONCLUSION: to the isolated Bacillus megaterium is capable of degrading tobacco straw partially and nicotine totally.


Asunto(s)
Bacillus megaterium/enzimología , Proteínas Bacterianas/metabolismo , Nicotiana/microbiología , Nicotina/metabolismo , Tallos de la Planta/microbiología , Microbiología del Suelo , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Bacillus megaterium/aislamiento & purificación , Proteínas Bacterianas/genética , Celulosa/metabolismo , Lignina/metabolismo , Datos de Secuencia Molecular , Filogenia , Tallos de la Planta/metabolismo , Nicotiana/metabolismo
13.
PLoS One ; 9(9): e108012, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25244184

RESUMEN

In this study, we isolated an endophytic quinclorac-degrading bacterium strain Q3 from the root of tobacco grown in quinclorac contaminated soil. Based on morphological characteristics, Biolog identification, and 16S rDNA sequence analysis, we identified strain Q3 as Bacillus megaterium. We investigated the effects of temperature, pH, inoculation size, and initial quinclorac concentration on growth and degrading efficiency of Q3. Under the optimal degrading condition, Q3 could degrade 93% of quinclorac from the initial concentration of 20 mg/L in seven days. We analyzed the degradation products of quinclorac using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The major degradation products by Q3 were different from those of previously identified quinclorac degrading strains, which suggests that Q3 may employ new pathways for quinclorac degradation. Our indoor pot experiments demonstrated that Q3 can effectively alleviate the quinclorac phytotoxicity in tobacco. As the first endophytic microbial that is capable of degrading quinclorac, Q3 can be a good bioremediation bacterium for quinclorac phytotoxicity.


Asunto(s)
Bacillus megaterium/metabolismo , Herbicidas/metabolismo , Quinolinas/metabolismo , Bacillus megaterium/clasificación , ADN Ribosómico/genética , Hidrólisis , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genética
14.
J Bacteriol ; 196(5): 1045-54, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24375103

RESUMEN

Previous work demonstrated that Bacillus megaterium QM B1551 spores that are null for the sleB and cwlJ genes, which encode cortex-lytic enzymes (CLEs), either of which is required for efficient cortex hydrolysis in Bacillus spores, could germinate efficiently when complemented with a plasmid-borne copy of ypeB plus the nonlytic portion of sleB encoding the N-terminal domain of SleB (sleB(N)). The current study demonstrates that the defective germination phenotype of B. megaterium sleB cwlJ spores can partially be restored when they are complemented with plasmid-borne ypeB alone. However, efficient germination in this genetic background requires the presence of sleL, which in this species was suggested previously to encode a nonlytic epimerase. Recombinant B. megaterium SleL showed little, or no, activity against purified spore sacculi, cortical fragments, or decoated spore substrates. However, analysis of muropeptides generated by the combined activities of recombinant SleB and SleL against spore sacculi revealed that B. megaterium SleL is actually an N-acetylglucosaminidase, albeit with apparent reduced activity compared to that of the homologous Bacillus cereus protein. Additionally, decoated spores were induced to release a significant proportion of dipicolinic acid (DPA) from the spore core when incubated with recombinant SleL plus YpeB, although optimal DPA release required the presence of endogenous CLEs. The physiological basis that underpins this newly identified dependency between SleL and YpeB is not clear, since pulldown assays indicated that the proteins do not interact physically in vitro.


Asunto(s)
Bacillus megaterium/metabolismo , Proteínas Bacterianas/metabolismo , Esporas Bacterianas/fisiología , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica/fisiología , Hidrólisis
15.
Int J Biol Macromol ; 60: 253-61, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23748002

RESUMEN

A marine sponge-associated bacterium Bacillus megaterium MSBN04 was used for the production of polyhydroxybutyrate (PHB) under solid state culture (SSC). A central composite design (CCD) was employed to optimize the production medium and to find out the interactive effects of four independent variables, viz. tapioca industry waste, palm jaggery, horse gram flour and trace element solution on PHB production. The maximum yield of PHB 8.637 mg g(-1) of substrate (tapioca industry waste) was achieved from biomass 15.203 mg g(-1) of substrate, using statistically optimized medium. The horse gram flour (nitrogen source) and trace element solution were found to be critical control factors for PHB synthesis. The (1)H NMR analysis revealed that the polymer was a PHB monomer. PHB obtained from this study having high molecular weight (6.7×10(5) Da) with low polydispersity index (PDI) value (1.71) and produced PHB was used to synthesize PHB polymeric nanoparticles using solvent displacement approach. Therefore, B. megaterium MSBN04 is an ideal candidate that can be exploited biotechnologically for the commercial production of PHB under solid state culture.


Asunto(s)
Bacillus megaterium/metabolismo , Fermentación , Hidroxibutiratos/aislamiento & purificación , Hidroxibutiratos/metabolismo , Bacillus megaterium/clasificación , Bacillus megaterium/crecimiento & desarrollo , Biotecnología , Hidroxibutiratos/química , Residuos Industriales
16.
J Biosci Bioeng ; 116(3): 302-8, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23548274

RESUMEN

Twenty bacterial strains were examined on their ability to produce polyhydroxyalkanoates (PHA) from different carbon sources under rich and depleted nitrogen conditions. Preliminary experiments with glucose as sole carbon source allowed to select PHA producing bacteria using FTIR spectroscopy. They were further tested with eight additional carbon substrates including organic, fatty acids or sugars. PHA content and monomer composition of four chosen strains (Pseudomonas putida mt-2, Bacillus megaterium DSM 90 and DSM 509, Corynebacterium glutamicum DSM 20137) were assessed by gas chromatography techniques for two cultural conditions: during growth phase on a mineral medium (MM) and after transfer of cells on a fresh MM without nitrogen (MM-N). For several carbon substrates, substantial amounts of PHA (up to 53% of the cell dry weight: CDW) were already obtained in MM for C. glutamicum DSM 20137 and the two B. megaterium strains; after transfer in MM-N, PHA contents remained constant except for B. megaterium DSM 509 where PHA production increased whatever the carbon source. P. putida mt-2 synthesized PHA under deprived nitrogen conditions. Highest PHA accumulation reached 48 and 77% of CDW with octanoic acid as substrate in B. megaterium DSM 90 and P. putida mt-2, respectively. Surprisingly, an atypical metabolic shift was observed for B. megaterium DSM 509 cultivated with nearly all unrelated carbon sources: whereas short chain length PHA (scl-PHA) were synthesized in MM, medium chain length PHA (mcl-PHA) were produced after transfer of cells into MM-N supplemented with the same substrate.


Asunto(s)
Bacillus megaterium/metabolismo , Carbono/metabolismo , Nitrógeno/metabolismo , Polihidroxialcanoatos/biosíntesis , Bacillus megaterium/clasificación , Bacillus megaterium/crecimiento & desarrollo , Corynebacterium glutamicum/metabolismo , Pseudomonas putida/metabolismo
17.
Bioprocess Biosyst Eng ; 36(10): 1463-74, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23340799

RESUMEN

This study demonstrated the improved polyhydroxybutyrate (PHB) production via high cell density cultivation of Bacillus megaterium BA-019 with balanced initial total sugar concentration and carbon to nitrogen (C/N) weight ratio. In the 10 L stirred fermentor operated at 30 °C, pH 7.0, 600 rpm, and 1.0 vvm air, with the initial total sugar concentration of 60 g/L and urea at the C/N weight ratio of 10:1, 32.48 g/L cell biomass with the corresponding PHB weight content of 26.94 % and volumetric productivity of 0.73 g/L h were obtained from batch cultivation. Continuing cultivation by intermittent feeding of the sugarcane molasses along with urea at the C/N weight ratio of 12.5:1 gave much improved biomass and PHB production (90.71 g/L biomass with 45.84 % PHB content and 1.73 g/L h PHB productivity). Similar biomass and PHB yields were obtained in the 90 L stirred fermentor when using the impeller tip speed as the scale-up criterion.


Asunto(s)
Bacillus megaterium/metabolismo , Fermentación , Hidroxibutiratos/metabolismo , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Secuencia de Bases , Biomasa , Carbono/metabolismo , Datos de Secuencia Molecular , Nitrógeno/metabolismo , Filogenia , ARN Ribosómico 16S/genética , Saccharum/metabolismo
18.
J Basic Microbiol ; 51(6): 614-24, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21780140

RESUMEN

An alkaline serine protease produced by Bacillus megaterium RRM2 isolated from the red alga, Kappaphycus alvarezii (Doty) Doty ex Silva was studied for the first time and the same analyzed for the production of protease in the present study. Identification of the bacterium was done on the basis of both biochemical analysis and by 16S rDNA sequence analysis. The extracellular protease obtained from B. megaterium RRM2 was purified by a three-step process involving ammonium sulphate precipitation, gel filtration (Sephadex G100) and Q-Sepharose column chromatography. The purity was found to be 30.6-fold with a specific activity of 3591.5 U/mg protein with a molecular weight of 27 kDa. The metal ions Ca(2+), Mg(2+), K(+) and Na(+) marginally enhanced the activity of the purified enzyme while Hg(2+), Cu(2+), Fe(2+), CO(2+) and Zn(2+), had reduced the activity. The enzyme was found to be active in the pH range of 9.0-10.0 and remained active up to 60 °C. Phenyl Methyl Sulfonyl Fluoride (PMSF) inhibited the enzyme activity, thus, confirming that this enzyme is an alkaline serine protease. Likewise, DTT also inhibited the enzyme thus confirming the disulfide nature of the enzyme. The enzyme exhibited a high degree of tolerance to Sodium Dodecyl Sulphate (SDS). The partially purified protease when used as an additive in the commercial detergents was found to be a suitable source for washing clothes especially those stained with blood. Further, it showed good dehairing activity within a short duration in goat skin without affecting its collagen component.


Asunto(s)
Bacillus megaterium/enzimología , Biotecnología/métodos , Detergentes/metabolismo , Inhibidores Enzimáticos/metabolismo , Cabello/metabolismo , Serina Proteasas/aislamiento & purificación , Serina Proteasas/metabolismo , Bacillus megaterium/clasificación , Bacillus megaterium/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Fraccionamiento Químico , Cromatografía Liquida , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Activadores de Enzimas/metabolismo , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Metales/metabolismo , Datos de Secuencia Molecular , Peso Molecular , Filogenia , ARN Ribosómico 16S/genética , Rhodophyta/microbiología , Análisis de Secuencia de ADN , Serina Proteasas/química , Temperatura
19.
J Bacteriol ; 193(16): 4199-213, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21705586

RESUMEN

Bacillus megaterium is deep-rooted in the Bacillus phylogeny, making it an evolutionarily key species and of particular importance in understanding genome evolution, dynamics, and plasticity in the bacilli. B. megaterium is a commercially available, nonpathogenic host for the biotechnological production of several substances, including vitamin B(12), penicillin acylase, and amylases. Here, we report the analysis of the first complete genome sequences of two important B. megaterium strains, the plasmidless strain DSM319 and QM B1551, which harbors seven indigenous plasmids. The 5.1-Mbp chromosome carries approximately 5,300 genes, while QM B1551 plasmids represent a combined 417 kb and 523 genes, one of the largest plasmid arrays sequenced in a single bacterial strain. We have documented extensive gene transfer between the plasmids and the chromosome. Each strain carries roughly 300 strain-specific chromosomal genes that account for differences in their experimentally confirmed phenotypes. B. megaterium is able to synthesize vitamin B(12) through an oxygen-independent adenosylcobalamin pathway, which together with other key energetic and metabolic pathways has now been fully reconstructed. Other novel genes include a second ftsZ gene, which may be responsible for the large cell size of members of this species, as well as genes for gas vesicles, a second ß-galactosidase gene, and most but not all of the genes needed for genetic competence. Comprehensive analyses of the global Bacillus gene pool showed that only an asymmetric region around the origin of replication was syntenic across the genus. This appears to be a characteristic feature of the Bacillus spp. genome architecture and may be key to their sporulating lifestyle.


Asunto(s)
Bacillus megaterium/clasificación , Bacillus megaterium/genética , Genoma Bacteriano , Bacillus megaterium/metabolismo , Cromosomas Bacterianos , Flagelos/genética , Flagelos/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Variación Genética , Datos de Secuencia Molecular , Filogenia , Plásmidos , Especificidad de la Especie
20.
Microbiology (Reading) ; 157(Pt 3): 721-726, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21163842

RESUMEN

An efficient 3,4-dichloroaniline (3,4-DCA)-mineralizing bacterium has been isolated from enrichment cultures originating from a soil sample with a history of repeated exposure to diuron, a major metabolite of which is 3,4-DCA. This bacterium, Bacillus megaterium IMT21, also mineralized 2,3-, 2,4-, 2,5- and 3,5-DCA as sole sources of carbon and energy. These five DCA isomers were degraded via two different routes. 2,3-, 2,4- and 2,5-DCA were degraded via previously unknown dichloroaminophenol metabolites, whereas 3,4- and 3,5-DCA were degraded via dichloroacetanilide.


Asunto(s)
Compuestos de Anilina/metabolismo , Bacillus megaterium/aislamiento & purificación , Bacillus megaterium/metabolismo , Microbiología del Suelo , Compuestos de Anilina/química , Bacillus megaterium/clasificación , Bacillus megaterium/genética , Biodegradación Ambiental , Medios de Cultivo , Diurona/química , Diurona/metabolismo , Genes de ARNr , Herbicidas/química , Herbicidas/metabolismo , Isomerismo , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
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