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1.
Braz. j. microbiol ; 49(3): 452-462, July-Sept. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-951792

RESUMO

Abstract Exopolysaccharide (EPS) biopolymers produced by microorganisms play a crucial role in the environment such as health and bio-nanotechnology sectors, gelling agents in food and cosmetic industries in addition to bio-flocculants in the environmental sector as they are degradable, nontoxic. This study focuses on the improvement of EPS production through manipulation of different culture and environmental conditions using response surface methodology (RSM). Plackett-Burman design indicated that; molasses, yeast extract and incubation temperature are the most effective parameters. Box-Behnken RSM indicated that; the optimum concentration for each parameter was 12% (w/v) for molasses, 6 g/L yeast extract and 30 °C for incubation temperature. The most potent bacterial isolate was identified as Bacillus velezensis KY498625. After production, EPS was extracted, purified using DEAE-cellulose, identified using Fourier transform infrared (FTIR), gel permeation chromatography (GPC) and gas chromatography-mass spectroscopy (GC-MS). The result indicated that; it has molecular weight 1.14 × 105 D consisting of glucose, mannose and galactose.


Assuntos
Polissacarídeos Bacterianos/metabolismo , Bacillus/metabolismo , Polissacarídeos Bacterianos/isolamento & purificação , Polissacarídeos Bacterianos/química , Bacillus/química , Microbiologia Industrial , Espectroscopia de Infravermelho com Transformada de Fourier , Meios de Cultura/metabolismo , Meios de Cultura/química , Peso Molecular
2.
Braz. j. microbiol ; 49(2): 414-421, Apr.-June 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889234

RESUMO

Abstract Agricultural crops suffer many diseases, including fungal and bacterial infections, causing significant yield losses. The identification and characterisation of pathogenesis-related protein genes, such as chitinases, can lead to reduction in pathogen growth, thereby increasing tolerance against fungal pathogens. In the present study, the chitinase I gene was isolated from the genomic DNA of Barley (Hordeum vulgare L.) cultivar, Haider-93. The isolated DNA was used as template for the amplification of the ∼935 bp full-length chitinase I gene. Based on the sequence of the amplified gene fragment, class I barley chitinase shares 93% amino acid sequence homology with class II wheat chitinase. Interestingly, barley class I chitinase and class II chitinase do not share sequence homology. Furthermore, the amplified fragment was expressed in Escherichia coli Rosetta strain under the control of T7 promoter in pET 30a vector. Recombinant chitinase protein of 35 kDa exhibited highest expression at 0.5 mM concentration of IPTG. Expressed recombinant protein of 35 kDa was purified to homogeneity with affinity chromatography. Following purification, a Western blot assay for recombinant chitinase protein measuring 35 kDa was developed with His-tag specific antibodies. The purified recombinant chitinase protein was demonstrated to inhibit significantly the important phytopathogenic fungi Alternaria solani, Fusarium spp, Rhizoctonia solani and Verticillium dahliae compared to the control at concentrations of 80 µg and 200 µg.


Assuntos
Antifúngicos/farmacologia , Quitinases/farmacologia , Hordeum/enzimologia , Proteínas Recombinantes/metabolismo , Antifúngicos/química , Antifúngicos/isolamento & purificação , Western Blotting , Quitinases/química , Quitinases/genética , Quitinases/isolamento & purificação , Cromatografia de Afinidade , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Hordeum/genética , Peso Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Homologia de Sequência de Aminoácidos
3.
Vitae (Medellín) ; 25(1): 26-36, 2018. Ilustraciones
Artigo em Inglês | LILACS, COLNAL | ID: biblio-994930

RESUMO

Background: ß-glucans (1-3: 1-4) are soluble fibers applied to foods due to their technological properties (water binding capacity, viscosity, emulsification and stabilization) and their beneficial effects on health. The functional properties of ß-glucans can be lost during the extraction and purification processes. The high viscosity of ß-glucans is related to a high molecular weight and its physiological properties in the intestine. Therefore, to characterize the fiber after its extraction and purification is fundamental to understand its possible applications in foods. Objectives: characterize ß-glucans extracted (EßG) and compare them with three commercial ß-glucans (CßG-A, CßG-B and CßG-C) to identify its possible applications in foods and to evaluate if enzymatic purification affects molecular and structurally the ß-glucans. Methods: barley ß-glucans were extracted (EßG), characterized by chemical analyzes, rheological behavior, and color, and compared to three commercial ß-glucans samples. Then, the extract was purified and its structural and molecular characteristics were calculated. Results: EßG contained 64.38 ± 3.54% of ß-glucans, high starch contamination (12.70 ± 1.73%), high content of calcium (8894 mg/kg), pseudoplastic behavior, and dark color (L* = 52.77 ± 0.7). All commercial samples showed low starch contamination, lighter color, and Newtonian behavior. After purification starch and protein contamination decreased (0.85 ± 0.46% and 5.50 ± 0.12% respectively), increased the content of ßG (69.45 ± 0.81%) and increased brightness (L* = 92.60 ± 1.70). Purified ß-glucans (PßG) showed a molar weight of 690 ± 1.6 kDa and species with degree polymerization 3 (DP3) to 11 (DP11) were identified on the structure. Conclusions: EßG extracts before the purification presented a high viscosity and contamination. The enzymatic purification process was effective and allowed to maintain a high molar mass of PßG and its distinctive molecular structures (species with DP3 and DP4). The commercial samples CßG-A and CßG-B showed a low content of ß-glucans. Finally, CßG-C presented the best physicochemical and rheological properties for its subsequent application in food.


Antecedentes: los ß-glucanos (1-3: 1-4) son fibras solubles aplicadas a los alimentos debido a sus propiedades tecnológicas (capacidad de retención de agua, viscosidad, emulsificación y estabilización) y a sus efectos beneficiosos en la salud. Las propiedades funcionales de los ß-glucanos pueden perderse durante los procesos de extracción y purificación. La alta viscosidad de los ß-glucanos está relacionada con un alto peso molecular y con sus propiedades fisiológicas en el intestino. Por lo tanto, caracterizar la fibra después de su extracción y purificación es fundamental para comprender sus posibles aplicaciones en alimentos. Objetivos: caracterizar ß-glucanos extraídos (EßG) y compararlos con tres marcas comerciales (CßG-A, CßG-B y CßG-C) para identificar su futura aplicación en alimentos y evaluar si la purificación enzimática afecta molecular y estructuralmente los ß-glucanos. Métodos: se extrajeron ß-glucanos de cebada (EßG), caracterizados por análisis químicos, comportamiento reológico y color, y se compararon con tres muestras comerciales. Posteriormente, el extracto (EßG) se purificó y se identificaron sus características estructurales y su peso molecular. Resultados: EßG contenía 64.38 ± 3.54% de ß-glucanos, alta contaminación con almidón (12.70 ± 1.73%), alto contenido de calcio (8894 mg / kg), comportamiento pseudoplástico y color oscuro (L* = 52.77 ± 0.7). Todas las muestras comerciales mostraron una baja contaminación con almidón, color más claro y comportamiento newtoniano. Después de la purificación de EßG, la contaminación con almidón y proteína disminuyó (0.85 ± 0.46% y 5.50 ± 0.12%, respectivamente), aumentó el contenido de ßG (69.45 ± 0.81%) y aumentó su luminosidad (L* = 92.60 ± 1.70). Los ß-glucanos purificados (PßG) mostraron un peso molar de 690 ± 1,6 kDa y se identificaron en la estructura especies con grado de polimerización desde 3 (GP3) hasta 11 (GP11). Conclusiones: los EßG antes de la purificación presentaron alta viscosidad y contaminación. El proceso de purificación enzimática fue efectivo y permitió mantener una alta masa molar de la fibra y sus estructuras moleculares características (especies con GP3 y GP4). Las muestras comerciales CßG-A y CßG-B mostraron un bajo contenido de ß-glucanos. Finalmente, la CßG-C presentó las mejores propiedades fisicoquímicas y reológicas para su posterior aplicación en alimentos.


Assuntos
Humanos , beta-Glucanas , Viscosidade , Fibras na Dieta , Alimentos Integrais , Peso Molecular
4.
Electron. j. biotechnol ; 30: 24-32, nov. 2017. tab, ilus, graf
Artigo em Inglês | LILACS | ID: biblio-1021325

RESUMO

Background: Prosopis, or mesquite (Prosopis juliflora (Sw.) DC.), was introduced in Saudi Arabia several decades ago and is heavily used in street, roadside, and park plantations. It shows great adaptation to the prevailing climatic conditions such as high temperature, severe drought, and salinity and spreads naturally in many parts of the Kingdom. This research was conducted to isolate allergen proteins and biogenic amines from the pollen grains of P. juliflora genotypes in Saudi Arabia from two regions, namely Al-Qassim and Eastern regions. Results: The results showed that 18 different allergen proteins were detected in P. juliflora genotypes, with molecular weight ranging from 14 to 97 kDa. Moreover, P. juliflora genotypes from the two studied regions contained eight biogenic amines, namely histamine, tyramine, tryptamine, ß-phenylethylamine, butricine, codapherine, spermidine, and spermine. All genotypes from the Al-Qassim region were found to contain all eight amines, while in the Eastern region, histamine was absent in three genotypes, spermine was absent in six genotypes, and spermidine was absent in three genotypes. Genotypes B23, E20, and E21 had the lowest biogenic amine quantity. Conclusions: All identified proteins from mesquite trees from both regions (Eastern and Al-Qassim) cause allergies in patients who are sensitive to pollen grains. Bioamines, except histamine and tyramine, were recorded at varying concentrations in different genotypes.


Assuntos
Pólen/química , Aminas Biogênicas/isolamento & purificação , Alérgenos/isolamento & purificação , Prosopis , Proteínas de Plantas/isolamento & purificação , Histamina/isolamento & purificação , Tiramina/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Eletroforese em Gel de Poliacrilamida , Genótipo , Peso Molecular
5.
Electron. j. biotechnol ; 29: 7-12, sept. 2017. ilus, graf, tab
Artigo em Inglês | LILACS | ID: biblio-1016095

RESUMO

Background: DegP is a serine protease that specifically cleaves and refolds unfolding proteins in the periplasmic space of the cells. To date, there is no information regarding DegP from halophilic bacteria. Chromohalobacter salexigens BKL5 is a moderately halophilic bacterium that has the ability to grow in a media containing more than 15% salt. Therefore, the objectives of this work were to clone and overexpress DegP-encoding gene from C. salexigens BKL5 and characterize its biochemical properties. Results: DegP-encoding gene was overexpressed in Escherichia coli BL21(DE3) CodonPlus in an active form. SDS-PAGE analysis showed that the molecular weight of the recombinant DegP was 45 kDa. Size-exclusion chromatography analysis suggested that recombinant DegP was present in two multimeric states, hexameric and dodecameric, with molecular weights of 297.9 and 579.12 kDa, respectively. Both conformations were enzymatically active when casein was used as substrate for enzymatic assay. Circular dichroism analysis showed that recombinant DegP was composed of 0.21­0.29 helical content, which was comparable to the helical content in the crystal structure of E. coli DegP. The basic/acidic residue ratio of recombinant DegP was 0.56, which was slightly higher than that of DegP from extreme halophiles (average, 0.45) but significantly lower than that of DegP from nonhalophiles (average, 0.94). Conclusions: Recombinant DegP from C. salexigens BKL5 showed proteolytic activity when ß-casein was used as a substrate. In silico analysis indicated that recombinant DegP had characteristics similar to those of halophilic proteins depending on its amino acid composition.


Assuntos
Serina Endopeptidases/genética , Proteínas Periplásmicas/genética , Chromohalobacter/enzimologia , Proteólise , Proteínas de Choque Térmico/genética , Proteínas Recombinantes , Serina Endopeptidases/metabolismo , Caseínas , Cromatografia em Gel , Dicroísmo Circular , Clonagem Molecular , Proteínas Periplásmicas/metabolismo , Eletroforese em Gel de Poliacrilamida , Escherichia coli , Salinidade , Chromohalobacter/genética , Proteínas de Choque Térmico/metabolismo , Peso Molecular
6.
Acta cir. bras ; 31(12): 813-820, Dec. 2016. graf
Artigo em Inglês | LILACS | ID: biblio-837660

RESUMO

ABSTRACT PURPOSE: To evaluate the role of low molecular chitosan containing sepia ink (LMCS) in ethanol-induced (5 ml/kg) gastric ulcer in rats. METHODS: Animals were divided into four groups (n = 12): normal group (Normal), negative control group (Con), experiment group (LMCS) and positive control Omeprazole group (OMZ). Gastric empty rate was detected in the first 7 days. Rats were sacrificed at 7, 14 and 21 day for histology and ELISA detections. RESULTS: Gastric empty was no significant differences among the groups (P > 0.05). Histological observation showed gastric mucosal LMCS treated had better healing effect. Hydroxyproline (Hyp) was significantly increased from 7 day (P < 0.05). LMCS significantly inhibited malondialdehyde (MDA) generation for lipid peroxidation from 7 day (P < 0.05). LMCS significantly promoted the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) at the earlier stage (P < 0.05). OMZ had the similar effects above. As for myeloperoxidase (MPO), LMCS significantly decreased and restored it to normal levels from 7 day (P < 0.05), it is earlier than OMZ which is from 14 day. CONCLUSION: LMCS can improve gastric mucosa tissue repair, exert significant influences on oxidative and antioxidant enzyme activities and neutrophil infiltration.


Assuntos
Animais , Ratos , Úlcera Gástrica/tratamento farmacológico , Quitosana/uso terapêutico , Sepia/química , Mucosa Gástrica/efeitos dos fármacos , Antiulcerosos/uso terapêutico , Antioxidantes/farmacologia , Úlcera Gástrica/induzido quimicamente , Distribuição Aleatória , Quitosana/química , Modelos Animais de Doenças , Etanol , Mucosa Gástrica/patologia , Hidroxiprolina/metabolismo , Tinta , Malondialdeído/metabolismo , Peso Molecular , Antioxidantes/metabolismo
7.
Braz. j. microbiol ; 47(3): 647-657, July-Sept. 2016. tab, graf
Artigo em Inglês | LILACS | ID: lil-788974

RESUMO

ABSTRACT The purpose of this study was to isolate, purify and optimize the production conditions of an organic solvent tolerant and thermostable lipase from Acinetobacter sp. AU07 isolated from distillery waste. The lipase production was optimized by response surface methodology, and a maximum production of 14.5 U/mL was observed at 30 ºC and pH 7, using a 0.5% (v/v) inoculum, 2% (v/v) castor oil (inducer), and agitation 150 rpm. The optimized conditions from the shake flask experiments were validated in a 3 L lab scale bioreactor, and the lipase production increased to 48 U/mL. The enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography and the overall yield was 36%. SDS-PAGE indicated a molecular weight of 45 kDa for the purified protein, and Matrix assisted laser desorption/ionization time of flight analysis of the purified lipase showed sequence similarity with GDSL family of lipases. The optimum temperature and pH for activity of the enzyme was found to be 50 ºC and 8.0, respectively. The lipase was completely inhibited by phenylmethylsulfonyl fluoride but minimal inhibition was observed when incubated with ethylenediaminetetraacetic acid and dithiothreitol. The enzyme was stable in the presence of non-polar hydrophobic solvents. Detergents like SDS inhibited enzyme activity; however, there was minimal loss of enzyme activity when incubated with hydrogen peroxide, Tween 80 and Triton X-100. The kinetic constants (Km and Vmax) revealed that the hydrolytic activity of the lipase was specific to moderate chain fatty acid esters. The Vmax, Km and Vmax/Km ratio of the enzyme were 16.98 U/mg, 0.51 mM, and 33.29, respectively when 4-nitrophenyl palmitate was used as a substrate.


Assuntos
Compostos Orgânicos , Solventes , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/biossíntese , Acinetobacter/enzimologia , Lipase/isolamento & purificação , Lipase/biossíntese , Compostos Orgânicos/química , Solventes/química , Especificidade por Substrato , Temperatura , Proteínas de Bactérias/química , Estabilidade Enzimática , Cinética , Cromatografia por Troca Iônica , Ativação Enzimática , Espaço Extracelular/enzimologia , Concentração de Íons de Hidrogênio , Íons , Lipase/química , Lipólise , Metais , Peso Molecular
8.
Braz. j. microbiol ; 47(1): 143-149, Jan.-Mar. 2016. tab, graf
Artigo em Inglês | LILACS | ID: lil-775118

RESUMO

Abstract Enzyme production by Aspergillus terreus NCFT 4269.10 was studied under liquid static surface and solid-state fermentation using mustard oil cake as a substrate. The maximum lipase biosynthesis was observed after incubation at 30 °C for 96 h. Among the domestic oils tested, the maximum lipase biosynthesis was achieved using palm oil. The crude lipase was purified 2.56-fold to electrophoretic homogeneity, with a yield of 8.44%, and the protein had a molecular weight of 46.3 kDa as determined by SDS-PAGE. Enzyme characterization confirmed that the purified lipase was most active at pH 6.0, temperature of 50 °C, and substrate concentration of 1.5%. The enzyme was thermostable at 60 °C for 1 h, and the optimum enzyme–substrate reaction time was 30 min. Sodium dodecyl sulfate and commercial detergents did not significantly affect lipase activity during 30-min incubation at 30 °C. Among the metal ions tested, the maximum lipase activity was attained in the presence of Zn2+, followed by Mg2+ and Fe2+. Lipase activity was not significantly affected in the presence of ethylenediaminetetraacetic acid, sodium lauryl sulfate and Triton X-100. Phenylmethylsulfonyl fluoride (1 mM) and the reducing, β-mercaptoethanol significantly inhibited lipase activity. The remarkable stability in the presence of detergents, additives, inhibitors and metal ions makes this lipase unique and a potential candidate for significant biotechnological exploitation.


Assuntos
Aspergillus/enzimologia , Lipase/metabolismo , Cátions Bivalentes/metabolismo , Eletroforese em Gel de Poliacrilamida , Estabilidade Enzimática , Ativadores de Enzimas/análise , Inibidores Enzimáticos/análise , Concentração de Íons de Hidrogênio , Lipase/química , Lipase/isolamento & purificação , Peso Molecular , Mercaptoetanol/metabolismo , Metais/metabolismo , Temperatura
9.
Braz. j. microbiol ; 46(4): 1147-1154, Oct.-Dec. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-769668

RESUMO

Abstract Pseudomonas aeruginosa, in spite of being a ubiquitous organism (as it is found in soil, water, and humans), is also an opportunistic pathogen. In order to maintain its diversity in the community, it produces various toxic proteins, known as, bacteriocins. In the present study, pyocin SA189, which is a bacteriocin produced by P. aeruginosa SA189 (isolated from a clinical sample) was characterized. P. aeruginosa SA189, as identified by the conventional and 16S rRNA gene amplification, produced pyocin SA189 of molecular weight of 66 k Da. The pyocin showed antimicrobial activity against several clinically relevant Gram-positive and Gram-negative bacteria and was substantially stable for wide ranges of temperature and pH. Furthermore, the pyocin also retained its biological activity upon treatment with metal ions, organic solvents, and various proteolytic and lipolytic enzymes. The data from the growth kinetics indicated that the maximum bacteriocin production occurred in the late log phase. Overall, our results signify the potential of pyocin SA189 as a bio-control agent.


Assuntos
Pseudomonas aeruginosa/metabolismo , Piocinas/isolamento & purificação , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Especificidade de Hospedeiro , Concentração de Íons de Hidrogênio , Peso Molecular , Pseudomonas aeruginosa/genética , Piocinas/química , /genética , Análise de Sequência de DNA , Temperatura
10.
Braz. j. microbiol ; 46(2): 425-432, Apr-Jun/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-749712

RESUMO

The strain JPL-2, capable of degrading fenoxaprop-P-ethyl (FE), was isolated from the soil of a wheat field and identified as Rhodococcus ruber. This strain could utilize FE as its sole carbon source and degrade 94.6% of 100 mg L−1 FE in 54 h. Strain JPL-2 could also degrade other aryloxyphenoxy propanoate (AOPP) herbicides. The initial step of the degradation pathway is to hydrolyze the carboxylic acid ester bond. A novel esterase gene feh, encoding the FE-hydrolyzing carboxylesterase (FeH) responsible for this initial step, was cloned from strain JPL-2. Its molecular mass was approximately 39 kDa, and the catalytic efficiency of FeH followed the order of FE > quizalofop-P-ethyl > clodinafop-propargyl > cyhalofop-butyl > fluazifop-P-butyl > haloxyfop-P-methyl > diclofop-methy, which indicated that the chain length of the alcohol moiety strongly affected the hydrolysis activity of the FeH toward AOPP herbicides.


Assuntos
Carboxilesterase/genética , Carboxilesterase/metabolismo , Herbicidas/metabolismo , Oxazóis/metabolismo , Propionatos/metabolismo , Rhodococcus/isolamento & purificação , Rhodococcus/metabolismo , Biotransformação , Clonagem Molecular , Análise por Conglomerados , Carboxilesterase/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Peso Molecular , Filogenia , /genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Rhodococcus/enzimologia , Rhodococcus/genética , Análise de Sequência de DNA , Microbiologia do Solo , Especificidade por Substrato , Triticum/crescimento & desenvolvimento
11.
Braz. j. microbiol ; 46(1): 251-260, 05/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-748253

RESUMO

An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The KM for sodium phytate hydrolysis was 30.9 mM, while the kcat and kcat/KM were 1.46 ×105 s−1 and 4.7 × 106 s−1.M−1, respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg2+, Cd2+, K+ and Ca2+, and it was drastically inhibited by F−. The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.


Assuntos
/isolamento & purificação , /metabolismo , Aspergillus niger/enzimologia , /química , Precipitação Química , Cromatografia em Gel , Cromatografia por Troca Iônica , Eletroforese em Gel de Poliacrilamida , Estabilidade Enzimática , Inibidores Enzimáticos/análise , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Peso Molecular , Multimerização Proteica , Proteólise , Peptídeo Hidrolases/metabolismo , Ácido Fítico/metabolismo , Especificidade por Substrato , Temperatura , Ultrafiltração
12.
Braz. j. microbiol ; 46(1): 285-292, 05/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-748256

RESUMO

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.


Assuntos
Aspergillus flavus/efeitos dos fármacos , Aspergillus flavus/enzimologia , Cobre/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Lacase/biossíntese , Ativação Transcricional/efeitos dos fármacos , Aspergillus flavus/genética , Aspergillus flavus/isolamento & purificação , Cromatografia em Gel , Meios de Cultura/química , DNA Fúngico/genética , Eletroforese em Gel de Poliacrilamida , Resíduos Industriais , Lacase/química , Lacase/isolamento & purificação , Dados de Sequência Molecular , Peso Molecular , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Microbiologia do Solo , Análise Espectral , Purificação da Água
13.
Mem. Inst. Oswaldo Cruz ; 110(1): 114-124, 03/02/2015. tab
Artigo em Inglês | LILACS | ID: lil-741621

RESUMO

This paper presents, from the perspective of technological development and production, the results of an investigation examining 61 clinical studies with vaccines conducted in Brazil between 1938-2013, with the participation of the Oswaldo Cruz Institute (IOC) and the Oswaldo Cruz Foundation (Fiocruz). These studies have been identified and reviewed according to criteria, such as the kind of vaccine (viral, bacterial, parasitic), their rationale, design and methodological strategies. The results indicate that IOC and Fiocruz have accumulated along this time significant knowledge and experience for the performance of studies in all clinical phases and are prepared for the development of new vaccines products and processes. We recommend national policy strategies to overcome existing regulatory and financing constraints.


Assuntos
Animais , Ração Animal/efeitos adversos , Proteínas na Dieta/química , Modelos Biológicos , Proantocianidinas/química , Rúmen/metabolismo , Brassica rapa/química , Precipitação Química , Proteínas na Dieta/metabolismo , Fermentação , Fabaceae/efeitos adversos , Fabaceae/química , Frutas/efeitos adversos , Frutas/química , Estrutura Molecular , Peso Molecular , Concentração Osmolar , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proantocianidinas/efeitos adversos , Proantocianidinas/metabolismo , Ruminantes , Ribulose-Bifosfato Carboxilase/química , Ribulose-Bifosfato Carboxilase/metabolismo , Rúmen/microbiologia , Solubilidade , Estereoisomerismo , Soroalbumina Bovina/química , Soroalbumina Bovina/metabolismo
14.
Ciênc. saúde coletiva ; 20(1): 75-84, 01/2015. graf
Artigo em Inglês | LILACS | ID: lil-733155

RESUMO

This study sought to verify the records on file and the number of cases of attempted suicide among children and adolescents who were attended by Emergency Care health professionals in the municipality of Matozinhos, Minas Gerais, Brazil. Documentary and descriptive research was conducted, the data for which was collected by means of an investigation of Outpatient Records from 2008 to 2010. Of the 73,000 files evaluated, those dealing with cases of attempted suicide among children and adolescents between the age of 3 and 18 years were selected. It was revealed that the health professionals, particularly physicians and nurses, fail to register the cases appropriately, invalidating information about the problem and potential prevention measures. The conclusion reached was that underreporting and the discrepancy of the diagnoses which were not duly referred to the competent agencies require rethinking and reviewing medical practices, and taking a systematic and careful look to address the individual as a complex whole.


Neste estudo procurou-se verificar o registro e o número de casos de tentativa de suicídio entre crianças e adolescentes do município de Matozinhos, Minas Gerais, Brasil, que foram atendidos pelos profissionais de saúde do Pronto-Atendimento. Trata-se de uma pesquisa documental e descritiva, cuja coleta dos dados ocorreu por meio de investigação nas Fichas Ambulatoriais, no período de 2008 a 2010. Das 73.000 fichas levantadas, selecionaram-se aquelas que tratavam de casos de tentativa de suicídio entre crianças e adolescentes do município, com idades entre três e 18 anos. Percebeu-se que os profissionais de saúde, mais especificamente os médicos e enfermeiros, não registram os casos de forma adequada, inviabilizando a informação sobre o problema e as medidas de prevenção. Concluiu-se que a subnotificação, a discrepância dos diagnósticos e o não encaminhamento aos órgãos competentes exigem repensar e rever a prática médica e dirigir um olhar sistematizado e cuidadoso para perceber o sujeito como um todo complexo.


Assuntos
Aldeídos/química , Citocromos c/química , Membranas Mitocondriais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Sequência de Aminoácidos , Cardiolipinas/química , Cardiolipinas/metabolismo , Citocromos c/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Concentração de Íons de Hidrogênio , Histidina/química , Histidina/metabolismo , Lisina/química , Lisina/metabolismo , Dados de Sequência Molecular , Peso Molecular , Estresse Oxidativo/fisiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fatores de Tempo
15.
Braz. j. phys. ther. (Impr.) ; 18(6): 538-543, 09/01/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-732351

RESUMO

BACKGROUND: The adapted arcometer has been validated for use in adults. However, its suitability for use in children can be questioned given the structural differences present in these populations. OBJECTIVE: To verify the concurrent validity, repeatability, and intra- and inter-reproducibility of the adapted arcometer for the measurement of the angles of thoracic kyphosis and lumbar lordosis in children. METHOD: Forty children were evaluated using both sagittal radiography of the spine and the adapted arcometer. The evaluations using the arcometer were carried out by two trained evaluators on two different days. In the statistical treatment, the intraclass correlation coefficient (ICC), Pearson's product moment correlation, Spearman's rho, the paired t test, and Wilcoxon's test were used (α=.05). RESULTS: A moderate and significant correlation was found between the x-ray and the adapted arcometer regarding thoracic kyphosis, but no correlation was found regarding lumbar lordosis. Repeatability and intra-evaluator reproducibility of the thoracic kyphosis and lumbar lordosis were confirmed, which was not the case of inter-evaluator reproducibility. CONCLUSION: The adapted arcometer can be used to accompany postural alterations in children made by the same evaluator, while its use for diagnostic purposes and continued evaluation by different evaluators cannot be recommended. Further studies with the aim of adapting this instrument for use in children are recommended. .


Assuntos
Proteínas de Bactérias/análise , Glicoproteínas de Membrana/análise , Glicoproteínas de Membrana/genética , Prodigiosina/biossíntese , Serratia marcescens/metabolismo , Centrifugação com Gradiente de Concentração , Eletroforese em Gel de Poliacrilamida , Peso Molecular , Glicoproteínas de Membrana/biossíntese , Solubilidade , Sarcosina/análogos & derivados , Serratia marcescens/análise
16.
Rev. bras. enferm ; 67(6): 957-964, Nov-Dec/2014.
Artigo em Português | LILACS, BDENF - Enfermagem | ID: lil-732825

RESUMO

Objetivo: identificar as necessidades e as preocupações prioritárias, manifestadas pelos pais no desempenho do seu papel, em três etapas do ciclo vital: adolescência, idade produtiva e idade madura. Metodologia: estudo exploratório com abordagem qualitativa, desenvolvido com quatorze pais residentes em um município no extremo sul do Brasil. Os dados foram coletados entre maio e agosto de 2011, por meio de entrevista em profundidade. Através da técnica da análise textual discursiva e da matriz construída com base na teoria bioecológica de Bronfenbrenner, foram construídas três categorias: Necessidades/preocupações do pai, geradas em sua relação com o mundo do trabalho; Necessidades/preocupações que emergem da relação de cuidado com os filhos e Preocupações dos pais com relação ao futuro dos filhos. Conclusão: identificou-se que a preocupação com o futuro dos filhos foi apontada por pais de todas as faixas-etárias investigadas. .


Objective: this study aimed to identify priority needs and concerns expressed by fathers in the performance of their role in three stages of the life cycle: adolescence, productive age, and mature age. Methodology: this is an exploratory study with a qualitative approach, conducted with fourteen fathers residing in a municipality in the extreme south of Brazil. The data were collected between May and August 2011 by means of the in-depth interview. Through the technique of written discourse analysis and the array built upon Bronfenbrenner's bioecological theory, we obtained three categories: fathers' needs/concerns, generated in their relationship with the world of work; needs/concerns that emerged from the relationship of care with the children; and fathers' concerns about the future of the children. Conclusions: we identified that the concern with the future of the children was pointed out by fathers of all age groups investigated. .


Objetivo: identificar las necesidades y preocupaciones prioritarias, manifestadas por los padres en el desempeño de su función, en tres etapas del ciclo de vida: adolescencia, edad productiva y edad madura. Metodología: estudio exploratorio con abordaje cualitativo, desarrollado con catorce padres residentes en un municipio en el extremo sur de Brasil. Los datos fueran colectados entre mayo y agosto de 2011, a través de entrevistas en profundidad. A través de la técnica de análisis textual y discursiva e de la matriz construida basada en la teoria bioecologica de Bronfenbrenner, fueran construidas tres categorías: Necesidades/ preocupaciones de lo padre, generado en suya relación con el mundo de lo trabajo; Necesidades/preocupaciones que emergen de la relación de cuidado con hijos e preocupaciones de los padres con lo futuro de los hijos. Conclusión: Se identifico que la preocupación con el futuro de los hijos fue apuntado por los padres de todas las edades averiguadas. .


Assuntos
Coenzima A Ligases/isolamento & purificação , Fenilacetatos/metabolismo , Pseudomonas/enzimologia , Aminoácidos/análise , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Cromatografia por Troca Iônica , Coenzima A Ligases/biossíntese , Coenzima A Ligases/metabolismo , Eletroforese em Gel de Poliacrilamida , Cinética , Peso Molecular , Pseudomonas/crescimento & desenvolvimento , Especificidade por Substrato , Termodinâmica , Ultracentrifugação
17.
Braz. j. microbiol ; 45(4): 1293-1302, Oct.-Dec. 2014. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-741279

RESUMO

Xylanase (EC 3. 2. 1. 8), hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry) as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7th day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH4SO2 precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa) using SDS-PAGE.


Assuntos
Microbiologia do Solo , Trichoderma/classificação , Trichoderma/isolamento & purificação , Xilosidases/análise , Cromatografia Líquida , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Peso Molecular , Técnicas de Tipagem Micológica , Filogenia , /genética , Análise de Sequência de DNA , Trichoderma/enzimologia , Trichoderma/crescimento & desenvolvimento , Xilosidases/química , Xilosidases/isolamento & purificação
18.
Salud pública Méx ; 56(6): 654-659, nov.-dic. 2014. tab
Artigo em Espanhol | LILACS | ID: lil-733345

RESUMO

La listeriosis es una enfermedad transmitida por alimentos (ETA) y ocasionada por Listeria monocytogenes. La importancia de ésta se debe a su impacto clínico, la alta tasa de mortalidad y el efecto económico derivado de los brotes asociados con el consumo de alimentos. En México, las fallas en los sistemas de vigilancia epidemiológicos son causa de información imprecisa sobre la incidencia de la listeriosis y sobre su caracterización como ETA. En este trabajo se presentan datos referentes a la presencia de la bacteria en alimentos, reportes de casos de la enfermedad y patologías relacionadas con infección por L. monocytogenes. La falta de datos exactos sobre la importancia de esta bacteria plantea la necesidad de concientizar a las instancias correspondientes para definir estrategias de búsqueda intencionada de L. monocytogenes en alimentos y de la recopilación de información clínica precisa que permita conocer la importancia clínica y epidemiológica de la listeriosis en México.


Listeriosis is caused by Listeria monocytogenes, an important food-borne disease due to its clinical forms, high mortality rate, and the economic impact in both clinical and food production industries. In Mexico, the lack of epidemiological surveillance systems leads to the need of accurate data on the incidence of listeriosis and its association with food-borne disease. In this paper, we present data about the presence of this bacterium in food, reports related to clinical cases of listeriosis, and information of diseases in which L. monocytogenes may be involved. However, in most of these cases the etiology was not established. Given this, there's a need to inform and warn the appropriate entities, to define strategies for the mandatory search of L. monocytogenes through the whole food production chain and clinical suspects, for the epidemiological importance and control of listeriosis in Mexico.


Assuntos
Animais , Cisteína Endopeptidases/isolamento & purificação , Proteínas do Ovo/metabolismo , Precursores Enzimáticos/isolamento & purificação , Antimaláricos/farmacologia , Cromatografia em Gel , Cisteína Endopeptidases/metabolismo , Inibidores de Cisteína Proteinase/farmacologia , Eletroforese em Gel de Poliacrilamida , Ativação Enzimática , Gema de Ovo/química , Precursores Enzimáticos/metabolismo , Concentração de Íons de Hidrogênio , Leucina/análogos & derivados , Leucina/metabolismo , Peso Molecular , Ortópteros
20.
Braz. j. microbiol ; 45(3): 1007-1015, July-Sept. 2014. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-727032

RESUMO

In the present study, a bacterium isolated from Marcha- a herbal cake used as traditional starter culture to ferment local wine in North East India, was evaluated for bacteriocin like inhibitory substance production and was tested against six food borne/spoilage causing pathogens viz. Listeria monocytogenes MTCC 839, Bacillus subtilis MTCC 121, Clostridium perfringens MTCC 450, Staphylococcus aureus, Lactobacillus plantarum and Leuconostoc mesenteroides MTCC 107 by using bit/disc method followed by well diffusion method. The bacterial isolate was identified as Brevibacillus borstelensis on the basis of phenotypic, biochemical and molecular characteristics using 16Sr RNA gene technique. Bacteriocin like inhibitory substance produced by Brevibacillus borstelensis AG1 was purified by gel exclusion chromatography. The molecular mass of the Brevibacillus borstelensis AG1 was found to be 12 kDa. Purified bacteriocin like inhibitory substance of Brevibacillus borstelensis was further characterized by studying the effect of temperature, pH, proteolytic enzyme and stability. Bacteriocin like inhibitory substance was found to be thermostable upto 100 °C, active at neutral pH, sensitive to trypsin, and partially stable till third week of storage thus showing a bright prospective to be used as a potential food biopreservative.


Assuntos
Bacteriocinas/isolamento & purificação , Bacteriocinas/farmacologia , Brevibacillus/isolamento & purificação , Brevibacillus/metabolismo , Bactérias Gram-Positivas/efeitos dos fármacos , Bacteriocinas/química , Cromatografia em Gel , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Microbiologia de Alimentos , Índia , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Peso Molecular , Filogenia , Estabilidade Proteica/efeitos da radiação , /genética , Análise de Sequência de DNA , Temperatura
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