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1.
Rev. bras. epidemiol ; 18(1): 194-207, Jan-Mar/2015. tab
Artículo en Portugués | LILACS | ID: lil-736436

RESUMEN

OBJETIVO: Analisar os fatores pessoais associados à prevalência e duração dos benefícios auxílio-doença decorrentes de sinovite e tenossinovite (CID10 M65). MÉTODO: Estudo transversal referente aos benefícios auxílio-doença decorrentes de sinovite e tenossinovite concedidos pelo Instituto Nacional de Seguro Social aos empregados no Brasil em 2008. Dados sobre o ramo de atividade econômica (Classificação Nacional de Atividades Econômicas - CNAE divisão, classe), sexo, idade, espécie e duração dos benefícios foram coletados do Sistema Único de Benefícios. A população corresponde à média mensal dos vínculos empregatícios declarados ao Cadastro Nacional de Informações Sociais. RESULTADOS: Em 2008 foram concedidos 35.601 benefícios auxílio-doença decorrentes de sinovite e tenossinovite, com prevalência de 10,9/10.000 vínculos empregatícios. No conjunto dos benefícios auxílio-doença houve maior razão de prevalência (RP) acidentária (RP 1,2), sendo esta maior em mulheres (RP 3,3), e em trabalhadores com idade acima de 39 anos (RP 1,4). As CNAE 37-Esgoto (55,4) e 60-Atividade de rádio e TV (47,1) apresentaram as maiores prevalências, no entanto, 64-Atividade de serviços financeiros e 6422-Bancos múltiplos caracterizaram mais acidentes de trabalho (RP 3,2 e 3,8, respectivamente) e maior duração (70 e 73 dias, respectivamente). A maior duração de benefício ocorreu entre trabalhadores com idade superior a 39 anos. Tanto a CNAE-divisão 60-Atividade de rádio e TV, quanto a CNAE-classe 6010-Atividade de rádio apresentaram elevadas razões de feminilidade (RP 8,1 e 10,8, respectivamente). CONCLUSÃO: A incapacidade para o trabalho por sinovite e tenossinovite apresenta associação tanto da prevalência quanto da duração com o ramo de atividade, sexo, idade e espécie de benefício (previdenciário/acidentário). .


OBJECTIVE: To analyse the personal and occupational factors associated with the prevalence and duration of sickness benefit claims due to synovitis and tenosynovitis (CID10 M65). METHODS: Cross-sectional study regarding sickness benefit claims due to synovitis and tenosynovitis granted to employees by National Institute of Social Security in Brazil in 2008. Data on economic activity (Economic Activities National Classification - CNAE division, class), sex, age, type and duration of benefits were collected from the Unified Benefit System. The study's population consists of the average monthly employment contracts declared to the National Register of Social Information. RESULTS: In 2008, 35,601 employees were granted sickness benefits due to synovitis and tenosynovitis, with a prevalence of 10.9/10,000 employments. Sickness benefits showed higher prevalence rates (PR) for work-related claims (PR 1,2), mostly made by females (PR 3.3) and by workers older than 39 years (PR 1,4). The CNAE 37-Sewage (55.4) and 60-Broadcasting Activity (47.1) had the highest overall prevalence. However, the 64-Financial service activities, except insurance and pension funding and 6422-Multiple banks with commercial service had the highest rates of work-related claims (RP 3.2 and 3.8, respectively), and the longer duration (70 and 73 days, respectively). Workers older than 39 years had the highest durations of work disability claims. Both the CNAE-division 60-Broadcasting Activity, and the CNAE-class 6010-Radio showed a high activity ratio of females (PR 8.1 and 10.8, respectively). CONCLUSION: The work disability due to synovitis and tenosynovitis presents prevalence and duration associated with economic activity, sex, age and kind of benefit (non work-related and work-related claims). .


Asunto(s)
Humanos , Globinas/química , Peróxido de Hidrógeno/química , Proteínas del Tejido Nervioso/química , Nitritos/química , Secuencia de Aminoácidos , Sitios de Unión , Catálisis , Cisteína/química , Cisteína/metabolismo , Disulfuros/química , Disulfuros/metabolismo , Globinas/metabolismo , Peróxido de Hidrógeno/metabolismo , Cinética , Modelos Moleculares , Datos de Secuencia Molecular , Mioglobina/química , Mioglobina/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Nitritos/metabolismo , Oxidación-Reducción , Conformación Proteica , Fenol/química , Fenoles/química , Fenilacetatos/química , Espectrometría de Masas en Tándem
2.
Indian J Biochem Biophys ; 2013 Jun; 50(3): 221-226
Artículo en Inglés | IMSEAR | ID: sea-147306

RESUMEN

The kinetic mechanism of enzymatic modification of flavonol quercetin with L-cysteine by horseradish peroxidase (HRP) was studied. Reaction of modification of quercetin was followed by recording spectral changes over time at 380 nm. All reactions were performed in 100 mM phosphate buffer pH, 6.0 at 20ºC. Kinetic parameters were determined from graphics of linear Michaelis-Menten equation. The values obtained at specified intervals were: Vmax = 0.17 ÷ 0.91 ΔA380/min, Km = 0.023 ÷ 0.5 mM, kcat = 0.21 ÷ 1.14 ΔA380/min∙nM-1 and Vmax/Km = 0.83 ÷ 26.55 ΔA380/min∙mM-1. It was found that all investigated reactions of the modification of quercetin with L-cysteine by HRP followed an ordered mechanism. We propose that HRP initially reacts with H2O2 than with quercetin and finally with L-cysteine, leading to the introduction of L-cysteine in the structure of quercetin.


Asunto(s)
Simulación por Computador , Cisteína/química , Activación Enzimática , Peroxidasa de Rábano Silvestre/química , Cinética , Modelos Químicos , Quercetina/química
3.
Arq. bras. med. vet. zootec ; 65(2): 430-438, abr. 2013. tab
Artículo en Portugués | LILACS | ID: lil-673118

RESUMEN

Foram utilizados ejaculados (n=25) de garanhões para avaliar o efeito de glutationa peroxidase (GPx) e cisteína na viabilidade de espermatozoides congelados. O sêmen foi diluído em Botu Crio, com antioxidantes, e foram formados os grupos: G1, Controle; G2, 1U GPx ; G3, 5U GPx; G4, 0,5mM cisteína; G5, 1mM cisteína. Depois foi envasado em palhetas (0,5mL) e congelado. Após descongelação, 37°C por 30 segundos, alíquotas foram analisadas quanto à integridade de membrana plasmática (IMP) e acrossoma (IAc), potencial de membrana mitocondrial (PMM) e cinética, nos tempos zero (T0) e 60 minutos (T60). GPx 5U e cisteína 0,5mM determinaram maior (P<0,05) IAc em T0 do que em T60. Cisteína 1mM resultou em maior (P<0,05) IAc em T60 do que GPx 1 e 5U e cisteína 0,5mM. O PMM de um garanhão no T60 foi mais alto (P<0,05) do que o de dois garanhões. VCL e VAP foram maiores (P<0,05) no T0 do que no T60 do grupo controle, e um garanhão apresentou, em geral, valores cinéticos mais altos (P<0,05) do que os demais. Conclui-se que a adição de glutationa peroxidase, nas concentrações de 1U e 5U, e de cisteína, nas concentrações de 0,5mM e 1mM, não interferem na integridade de espermatozoides criopreservados de equinos, mas preservam os parâmetros cinéticos de VCL e VAP após 60 minutos de incubação. Ressalta-se, ainda, que o garanhão tem uma forte influência nas características espermáticas pós-congelação.


Ejaculates (n=25) of horses were used to assess the effect of glutathione peroxidase (GPx) and cysteine on the viability of frozen sperm cells. Semen was extended at Botu Crio with antioxidants, and divided in groups: G1, control; G2, 1 U GPx; G3, 5U GPx; G4, 0.5mM cysteine and G5, 1mM cysteine, packed in 0.5mL straws, and frozen. After thawing (37° C for 30 seconds) samples were analyzed for plasma membrane (IMP) and acrosome integrity (IAc), mitochondrial membrane potential (MMP) and kinematic, at zero (T0) and 60 minutes after (T60). GPx 5U and cysteine 0.5mM increased (P<0.05) IAc at T0, when compared to T60. Cysteine 1mM resulted in a higher (P<0.05) IAc on T60, than GPx 1 and 5U, and cysteine 0.5mM. The PMM from a stallion on T60 was higher (P<0.05) than those of two stallions. In sperm kinematic, VCL and VAP were higher (P<0.05) at T0 compared to T60 for the control group, and one stallion showed larger (P<0.05) kinematic values than other animals. It is concluded that the addition of glutathione peroxidase at concentrations 1U and 5U, and cysteine, at concentrations of 0.5mM and 1mM, does not interfere with the integrity of cryopreserved equine sperm, but preserves the kinetic parameters VCL and VAP after 60 minutes of incubation. It should be noted also that the stallion has a strong influence on sperm characteristics post-freezing.


Asunto(s)
Animales , Análisis de Semen/métodos , Cisteína/química , Glutatión Peroxidasa , Caballos/clasificación , Criopreservación/instrumentación
4.
Clinics ; 67(10): 1171-1179, Oct. 2012. ilus, tab
Artículo en Inglés | LILACS | ID: lil-653481

RESUMEN

OBJECTIVE: To explore the use of β-lactoglobulin polymerized using microbial transglutaminase and heating to identify whether protein polymerization could reduce in vivo allergenicity and maintain in vitro and ex vivo immunoreactivity for use in tolerance-induction protocols. METHODS: Based on previous protocols applied in mice and children, we performed in vivo challenges (using a skin prick test) with native and polymerized β-lactoglobulin in adult patients with an IgE-mediated allergy to plactoglobulin. In vitro humoral immunoreactivity was analyzed using immunoblotting. Cell-mediated immunoreactivity was analyzed using ex vivo challenges with native and polymerized β-lactoglobulin and monitored by leukocyte adherence inhibition tests. RESULTS: The skin tests demonstrated that there was a significant reduction in immediate cutaneous reactivity after polymerization. Polymerization did not decrease the immunoblotting detection of s-IgE specific to β-lactoglobulin. Cell-mediated immunoreactivity, as assessed by ex vivo challenges and leukocyte adherence inhibition tests, did not exhibit significant differences between leukocytes challenged with native versus polymerized β-lactoglobulin. CONCLUSIONS: The polymerization of β-lactoglobulin decreased in vivo allergenicity and did not decrease in vitro humoral or ex vivo cell-mediated immunoreactivity. Therefore, we conclude that inducing polymerization using transglutaminase represents a promising technique to produce suitable molecules for the purpose of designing oral/ sublingual tolerance induction protocols for the treatment of allergies.


Asunto(s)
Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Cisteína/inmunología , Tolerancia Inmunológica/inmunología , Lactoglobulinas/inmunología , Hipersensibilidad a la Leche/inmunología , Transglutaminasas/inmunología , Alérgenos/inmunología , Estudios de Casos y Controles , Cisteína/química , Calefacción , Immunoblotting , Inmunoglobulina E/sangre , Prueba de Inhibición de Adhesión Leucocitaria , Hipersensibilidad a la Leche/prevención & control , Polimerizacion , Pruebas Cutáneas , Estadísticas no Paramétricas , Transglutaminasas/química
5.
Rev. colomb. psiquiatr ; 41(1): 217-228, ene.-abr. 2012. ilus, graf, tab
Artículo en Español | LILACS | ID: lil-639942

RESUMEN

Introducción: En psiquiatría de enlace se logra obtener una visión integral del tratamiento y de las necesidades de cada paciente prestando especial atención a las interacciones medicamentosas y a las contraindicaciones. Algunos casos particulares motivaron la descripción, reporte y revisión bibliográfica acerca de otras posibles aplicaciones de fármacos antagonistas de los recetores 5HT2A y 3, particularmente mirtazapina y olanzapina, en síndrome de hiperalgesia, tinitus y leucoencefalopatía multifocal progresiva por virus JC. Método: reporte de casos. Resultados y Conclusiones: Se describen los casos de tres pacientes en los cuales fue necesario usar mirtazapina y olanzapina no solo para el control de los síntomas psiquiátricos (afectivos, comportamentales y trastorno del sueño), sino también como coadyuvantes en las patologías de base de cada paciente. El uso de cualquier medicamento en psiquiatría de enlace debe tener en cuenta el contexto del paciente, la comorbilidad, las contraindicaciones y las interacciones farmacológicas para garantizar un desenlace positivo, además de promover el trabajo multidisciplinario entre especialistas.


Introduction: In liaison psychiatry it is possible to get an integral view of patient's treatment and needs, paying special attention to pharmacological interactions and contraindications. Some particular cases motivated the description, report and review about other possible applications of 5HT2A and 5HT3 antagonist, particularly Mirtazapine and Olanzapine, in hyperalgesia syndrome, tinnitus and Progressive Multifocal Leukoencephalopathy by JC virus. Method: Cases report. Results: We describe 3 cases of patients in which Mirtazapine and Olanzapine were necessary not only to control psychiatric symptoms (affective / behavioral symptoms and insomnia) but to act as adjuvant therapy in axis III diseases. The use of any drug in psychiatry must take in to account the context of the patient, the presence of comorbidity, contraindications and pharmacological interactions so as to grant a positive outcome also promoting the multidisciplinary work between specialists.


Asunto(s)
Humanos , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Núcleo Celular/metabolismo , Cisteína/metabolismo , Neuronas/metabolismo , Tiorredoxinas/metabolismo , Factores de Transcripción/metabolismo , Secuencias de Aminoácidos , Proteínas Adaptadoras Transductoras de Señales/genética , Línea Celular Tumoral , Cisteína/química , Citoplasma/metabolismo , Disulfuros/química , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Mutación , Neuronas/citología , Oxidación-Reducción , Mapeo de Interacción de Proteínas , Transducción de Señal , Transcripción Genética , Tiorredoxinas/genética , Factores de Transcripción/genética
6.
Indian J Biochem Biophys ; 2007 Jun; 44(3): 179-82
Artículo en Inglés | IMSEAR | ID: sea-26455

RESUMEN

Hydrogen sulphide (H2S), a signaling gasotransmitter and a potent vasorelaxant is endogenously produced by the enzymes cystathionine-beta-synthase (CBS) and cystathionine-gamma-lyase (CSE). CBS is a predominant source of H2S in the central nervous system, while CSE is the major H2S producing enzyme in the brain and other nervous tissues. Though the expression of these enzymes in the blood lymphocytes is known, H2S formation in the lymphocytes has not been reported so far. In the present study, H2S levels in the lymphocytes of healthy control subjects were estimated, after suitable modifications in a routine method [Stipanuk M H & Beck P W (1982) Biochem J 206, 267-277] used for detecting tissue levels of H2S. In this method, homocysteine (Hcys) due to its higher solubility was used as the substrate in place of L-cysteine and NaOH was used in place of zinc acetate to increase the entrapment of H2S in the central well. A mean H2S level of 11.64 +/- 6.36 microM/min/mg protein was detected in the lymphocytes of 8 subjects (mean age, 24 +/- 2; 2 male, 6 female). The modified method was found to be more sensitive for H2S estimation in human lymphocytes. As endogenous H2S is reported to be involved in the pathogenesis of various cardiovascular and pulmonary diseases, the levels of H2S in lymphocytes can be a marker of the endogenous tissue levels.


Asunto(s)
Adulto , Cisteína/química , Femenino , Homocisteína/química , Humanos , Sulfuro de Hidrógeno/análisis , Leucocitos Mononucleares/metabolismo , Linfocitos/citología , Masculino , Modelos Biológicos , Modelos Químicos , Transducción de Señal , Especificidad por Sustrato , Distribución Tisular , Acetato de Zinc/química
8.
Indian J Biochem Biophys ; 2006 Feb; 43(1): 37-40
Artículo en Inglés | IMSEAR | ID: sea-29086

RESUMEN

The kinetics and mechanism of the reduction of ferricytochrome c [Cyt c(III)] by substrates namely glutathione (GSH) and L-cysteine (L-cys) have been investigated spectrophotometrically employing [substrate]T > [Cyt c(III)]T. The reaction exhibits first order dependence in [substrate]T and [Cyt c(III)]T. The pseudo-first order rate constant increases with an increase in pH, indicating that the conjugate base form of the HCyt c(III) is a better oxidant than the parent HCyt c(III). The electron transfer rate constants between the oxidants and GSH for both the k1 and k2 paths are found to be greater than that with L-cysteine. Hence, GSH is a better reductant of Cyt c(III) as compared to L-cysteine. A suitable mechanism has been proposed on the basis of experimental findings. The deprotonation constant for HCyt c(III) and the second order rate constants of k1 and k2 paths for the present reaction at 25 degrees C have been determined.


Asunto(s)
Animales , Cisteína/química , Citocromos c/química , Glutatión/química , Caballos , Cinética , Oxidación-Reducción
9.
Egyptian Journal of Food Science. 2006; 34: 21-36
en Inglés | IMEMR | ID: emr-145496

RESUMEN

THE AMINO acids and vegetable proteins have been recommended as safe flavouring precursors of thermal process flavouring. Therefore the present work was designed to examine the effect of such compounds on the overall qualities of corn flavour extrudates. Amino acids [proline and cysteine], vegetable proteins [soybean and gluten] and sucrose were added to corn flour pre-extrusion process. A sample free from any additives was considered as control. The sensory evaluation for each sample was carried out, the expansion ratio and protein content were determined, the results showed a significant increase in all sensory attributes comparing with the control sample. Samples containing proline or cysteine showed high quality attributes whereas addition of soybean protein or gluten resulted in significant decrease in appearance and texture attributes compared with the sample containing sucrose only. An agreement between the sensory results and expansion ratio for all samples was observed for the protein content. On the other hand, the GC and GC-MS analysis confirmed the aroma quality of each sample


Asunto(s)
Cisteína/química , Glycine max/química , Glútenes , Sacarosa/química , Cromatografía de Gases/métodos , Aminoácidos/química , Proteínas/química
10.
Egyptian Journal of Hospital Medicine [The]. 2006; 23 (June): 287-308
en Inglés | IMEMR | ID: emr-145511

RESUMEN

Hair is an of the epidermis in mammals and consists of two large groups of human hair proteins. One is hard -keratins and the other is matrix proteins. The present investigation aimed to compare the ultrastructural of the hair scale using the scanning electron microscope, and the proteins and amino acids content of the keratin in seven mammalian species. The values of the hair thickness, x/y feret and hair pattern of the species in the present study confirm the presence of species-specific characteristics and ultra structural variation. The situation in man differs from the wild mammals due to damage of hair cuticle caused by mechanical abuse, exposure to ultraviolet radiation and chemical over processing. The maximum amount of extracted proteins from hair keratin was analyzed by SDS-PAGE. The electrophoretic patterns showed an overall degree of similarity. However, differences exist between species in the intensity of stain. Quantitatively, the electrophoretic patterns scanned and analyzed using gel protein analyzer. The results showed no difference between the molecular mass of some species, but different in molecular mass distribution. Amino acid composition of keratin of mammalian hair species of the present study showed some variation, especially for methionine, isoleucine, lysine and arginine. The other amino acids studied are significantly present in most hair. One of the later amino acid is cysteine. Cysteine is a very important due to the presence of disulfate cross-links


Asunto(s)
Queratinas/química , Proteínas de la Matriz Extracelular/química , Electroforesis/estadística & datos numéricos , Microscopía Electrónica de Rastreo , Metionina/química , Isoleucina/química , Lisina/química , Arginina/química , Cisteína/química
11.
Experimental & Molecular Medicine ; : 18-26, 2006.
Artículo en Inglés | WPRIM | ID: wpr-77905

RESUMEN

The constrained alpha-helical structure of a C-peptide is useful for enhancing anti-HIV-1 activity. The i and i+3 positions in an alpha-helical structure are located close together, therefore D-Cys (dC) and L-Cys (C) were introduced at the positions, respectively, to make a dC-C disulfide bond in 28mer C-peptides. Accordingly, this study tested whether a dC-C disulfide bond would increase the alpha-helicity and anti-HIV-1 activity of peptides. A C-peptide can be divided into three domains, the N-terminal hydrophobic domain (HPD), middle interface domain (IFD), and C-terminal hydrogen domain (HGD), based on the binding property with an N-peptide. In general, the dC-C modifications in HPD enhanced the anti-HIV-1 activity, while those in IFD and HGD resulted in no or much less activity. The modified peptides with no activity clearly showed much less alpha-helicity than the native peptides, while those with higher activity showed an almost similar or slightly increased alpha-helicity. Therefore, the present results suggest that the introduction of a dC-C bridge in the N-terminal hydrophobic domain of a C-peptide may be useful for enhancing the anti-HIV-1 activity.


Asunto(s)
Humanos , Secuencia de Aminoácidos , Fármacos Anti-VIH/síntesis química , Línea Celular , Dicroismo Circular , Cisteína/química , Disulfuros/química , Proteína gp41 de Envoltorio del VIH/química , VIH-1/efectos de los fármacos , Concentración 50 Inhibidora , Modelos Moleculares , Datos de Secuencia Molecular , Péptidos/síntesis química , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Relación Estructura-Actividad
12.
Journal of Korean Medical Science ; : 123-126, 2004.
Artículo en Inglés | WPRIM | ID: wpr-20641

RESUMEN

X-linked severe combined immunodeficiency (X-SCID) is a rare, life-threatening immune disorder, caused by mutations in the gamma c chain gene, which encodes an essential component of the cytokine receptors for interleukin-2 (IL-2), IL-4, IL-7, IL-9, IL-15, and IL-21. A 13-month-old boy with recurrent infections who had reduced serum immunoglobulin levels and decreased numbers of CD3, CD16/56 cells was evaluated for gamma c chain gene mutation and protein expression. The patient had a C-to-T point mutation at nucleotide position 690, one of the hot spots, resulting in a single amino acid substitution of cysteine for arginine (R226C), as determined by direct sequencing and PCR-RFLP. The patient's mother was a heterozygous carrier. Percutaneous umbilical cord blood sampling was performed at the 6-month of gestation in a subsequent pregnancy. As the immunophenotype of the fetus showed an identical pattern, the pregnancy was terminated and genetic analysis of the abortus confirmed recurrence. This is the first report of the molecular diagnosis of X-SCID in Korea. Genetic analysis of the gamma c chain gene is useful for definite diagnosis and genetic counseling for X-SCID.


Asunto(s)
Femenino , Humanos , Masculino , Arginina/química , Cisteína/química , ADN/metabolismo , Análisis Mutacional de ADN , Citometría de Flujo , Asesoramiento Genético/métodos , Heterocigoto , Inmunoglobulinas/metabolismo , Inmunofenotipificación/métodos , Corea (Geográfico) , Ligamiento Genético , Mutación , Linaje , Mutación Puntual , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Receptores Inmunológicos/genética , Análisis de Secuencia de ADN , Inmunodeficiencia Combinada Grave/diagnóstico , Factores de Tiempo , Cromosoma X
13.
Journal of Korean Medical Science ; : 856-860, 2002.
Artículo en Inglés | WPRIM | ID: wpr-125130

RESUMEN

A family with paramyotonia congenita (PC) is presented. At least 10 family members were affected in an autosomal dominant inheritance pattern. The proband had cold-sensitive muscle stiffness, paradoxical myotonia, and intermittent muscle weakness since childhood. The serum level of creatine kinase was mildly elevated and short exercise test with cooling revealed a drastic reduction of compound muscle action potentials with repetitive discharges. Muscle biopsy revealed marked variation in the fiber size and increased internal nuclei. The molecular biological study revealed a common missense mutation (Arg1448Cys) at the voltage-gated sodium channel gene (SCN4A). The repetitive CMAP discharges during short exercise test with cooling observed in the proband has not been reported previously. This observation needs to be confirmed among PC patients with different mutations. This is the first report on a PC family confirmed by the molecular biological technique in Korea.


Asunto(s)
Adulto , Femenino , Humanos , Masculino , Arginina/química , Núcleo Celular/metabolismo , Creatina Quinasa/sangre , Cisteína/química , Análisis Mutacional de ADN , Ejercicio Físico , Corea (Geográfico) , Mutación Missense , Trastornos Miotónicos/genética , Linaje , Fenotipo , Canales de Sodio/genética
14.
Indian J Biochem Biophys ; 2001 Jun; 38(3): 180-5
Artículo en Inglés | IMSEAR | ID: sea-28955

RESUMEN

Purification and characterization of an extracellular invertase produced by Aspergillus ochraceus TS are reported. The enzyme was purified (42-fold) from culture filtrate by salt precipitation, ion-exchange and gel filtration. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme showed a single band of molecular mass 66 kDa. The molecular mass of the native enzyme was found to be 130 kDa by gel filtration. The purity of the protein was also checked against its antiserum raised in rabbits by two-dimensional immunodiffusion in agarose gel and Western blot that showed a single band. It is a glycoprotein with mannose as its carbohydrate residue. The enzyme showed high affinity for sucrose with a Km of 3.5 mM. The amino acid analysis revealed a high proportion of acidic residues but it had a low content of cysteine, histidine and arginine comparable to other fungal invertases.


Asunto(s)
Aminoácidos/química , Arginina/química , Aspergillus/enzimología , Western Blotting , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Cisteína/química , Electroforesis en Gel de Agar , Electroforesis en Gel de Poliacrilamida , Glicósido Hidrolasas/química , Histidina/química , Concentración de Iones de Hidrógeno , Iones , Cinética , Manosa/química , Peso Molecular , Unión Proteica , Sacarosa/química , beta-Fructofuranosidasa
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