RESUMO
A simple, rapid and straightforward procedure for identification and determination of intracellular and extracellular activity of aminopeptidases employing synthetic substrates beta-naphtylamides of L-Ala, L-Phe, and L-Tyr was used. Poppy cells (Papaver somniferum L.) permeabilized by Tween 80 were immobilized via crosslinking by glutaraldehyde. Glutaraldehyde immobilized poppy cells lost their viability and demonstrated significantly lower aminopeptidase activities than untreated control cells probably due to a damage to the enzyme active centre. Poppy cells immobilized by pectate and alginate have retained high activity of studied aminopeptidases. The culture medium (without cells) used for the identification and determination of extracellular enzyme activities retained 20-21%, whereas intracellular activities were estimated to be 79-80% of total enzyme activity. Thus the intracellular specific activity was 1.00-1.07 higher.
Assuntos
Alanina/análogos & derivados , Alanina/metabolismo , Aminopeptidases/metabolismo , Papaver/enzimologia , Fenilalanina/análogos & derivados , Fenilalanina/metabolismo , Tirosina/análogos & derivados , Tirosina/metabolismo , Alanina/química , Alginatos/química , Amidas/química , Amidas/metabolismo , Aminopeptidases/química , Permeabilidade da Membrana Celular , Sobrevivência Celular , Células Cultivadas , Espaço Extracelular/enzimologia , Glucose/metabolismo , Glutaral/química , Espaço Intracelular/enzimologia , Pectinas/química , Fenilalanina/química , Polissorbatos , Tirosina/químicaRESUMO
A simple and rapid procedure for the identification and determination of extracellular invertase from a culture medium of tomato cell suspension cultures is described. Sucrose was used as substrate for the determination of the extracellular and intracellular activities of the enzyme. The culture medium (without cells) was used for identification and determination of extracellular enzyme activity. Intracellular activity was estimated from the cell suspension.
Assuntos
Solanum lycopersicum/enzimologia , beta-Frutofuranosidase/metabolismo , Células Cultivadas , Meios de Cultura , Espaço Extracelular/enzimologia , Solanum lycopersicum/citologia , Sacarose/metabolismo , beta-Frutofuranosidase/químicaRESUMO
A simple, rapid and reproducible procedure for the identification of extracellular Californian poppy (Eschscholzia californica Cham.) beta-galactosidase is described using callus cultures of seedlings from the tested plant, roots of 4-days-old seedlings of Californian poppy germinating on agar plates and cell suspension cultures cultivated from callus cultures. 6-Bromo-2-naphthyl-beta-D-galactopyranoside and p-nitrophenyl-beta-D-galactopyranoside were used as substrates for the determination of the intracellular and extracellular activities of beta-galactosidase. The extracellular beta-galactosidase activity was identified by evaluating the dye-zones in an agar medium. The enzyme from Californian poppy callus cultures or from seedling roots cultivated on agar plates supplemented with 6-bromo-2-naphthyl-galactopyranoside hydrolyzed this substrate releasing 6-bromo-2-naphthol. By simultaneous coupling with hexazonium p-rosaniline the corresponding (reddish-brown) azo-dye was formed. The agar plate method described permits rapid, simple and specific detection of plant producers of extracellular beta-galactosidase.
Assuntos
Plantas/enzimologia , beta-Galactosidase/análise , Células Cultivadas , Espaço Extracelular/enzimologia , Galactosídeos/química , Indicadores e Reagentes , Papaver/química , Reprodutibilidade dos TestesRESUMO
The present paper aims to inform the pharmaceutical public about anthracenons and their natural producers. The first available mention dates from 1888 and the present authors also describe their synthesis and use in clinical practice.
Assuntos
Antracenos/química , Extratos Vegetais , Animais , Antracenos/uso terapêutico , Antracenos/toxicidade , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/uso terapêutico , Humanos , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Extratos Vegetais/toxicidadeRESUMO
A simple, sensitive and reproducible method of detection of extracellular beta-galactosidase was worked out. beta-Galactosidase secreted by the callus culture, or the roots of sprouting plants, hydrolyzes the substrate (6-bromo-2-naphthyl-beta-D-galactopyranoside) to produce beta-D-galactose and 6-bromo-2-naphthol, which after simultaneous azocoupling with hexazotized p-rosaniline, or basic fuchsine, produce a reddish brown compound, nearly insoluble in water (a diazonium salt). The colouring under the callus culture and around it, as well as the colouring of the roots of the sprouting plants, is a manifestation of the activity of extracellular beta-galactosidase by the objects under study. The intensity of the colouring is a measure of their enzymatic activity. The share of intracellularly localized enzyme represents the majority and the share of extracellular beta-galactosidase the minority.
Assuntos
Plantas/enzimologia , beta-Galactosidase/biossíntese , Raízes de PlantasRESUMO
Curcuma vera is one of the medicinal plants of traditional Chinese medicine. The plant is the natural source of one of natural antioxidative agents--curcumin. The present paper summarizes information about the properties of curcumin and presents the characteristics of Curcuma vera, its producer.
Assuntos
Antioxidantes , Curcumina , Antioxidantes/química , Antioxidantes/uso terapêutico , Curcumina/química , Curcumina/uso terapêutico , HumanosRESUMO
Cell suspensions of gherkin (Cucumis sativus L.) were permeabilized by Tween-80, and immobilized by glutaraldehyde. Beta-galactosidase showed pH optimum at 4.9 and temperature optimum at 58 degrees C. The enzyme catalysed hydrolysis was linear for 3 h with 60-68% conversion of the substrate. The cells characterized by high beta-galactosidase activity and stability on long-term storage showed valuable technological properties.
Assuntos
Cucumis sativus/enzimologia , beta-Galactosidase/metabolismo , Catálise , Células Imobilizadas , Estabilidade Enzimática , Hidrólise , beta-Galactosidase/análiseRESUMO
Papaver somniferum L., (opium poppy) cells were after permeabilization in Tween 80 immobilized by glutaraldehyde without any carrier. Cells immobilized by cross-linking performed the hydrolysis of sucrose. The immobilized cells were characterized by high invertase activity and appropriate physico-mechanical properties.