Cell suspension of Papaver somniferum L. (opium poppy) was permeabilized by Tween 80 and immobili... more Cell suspension of Papaver somniferum L. (opium poppy) was permeabilized by Tween 80 and immobilized by glutaraldehyde. The highest saccharase activity was at pH 4.6 and 50°C. The hydrolysis of the substrate was linear for 5 h reaching 69% of conversion. A very good storage stability was achieved when using dry catalyst, or a solution of 0.15 M NaCl with the addition of chloramphenicol, (1-methyldodecyl)-dimethylamin-4-oxide (ATDNO), chlortetracycline hydrochloride (CLCTC) or by freezing the immobilized cells in 0.15 M NaCl. The cells characterized by high enzyme activity and stability in long-term storage showed convenient technological and physicomechanical properties.
A simple and rapid procedure for the identification and determination of extracellular invertase ... more 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.
Ceská a Slovenská farmacie : casopis Ceské farmaceutické spolecnosti a Slovenské farmaceutické spolecnosti, 2000
A simple, sensitive and reproducible method of detection of extracellular beta-galactosidase was ... more 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.
Chemistry of Natural Compounds - CHEM NAT COMPD, 2002
A simple, rapid, and sensitive procedure for the identification of plant extracellular a- and ß-g... more A simple, rapid, and sensitive procedure for the identification of plant extracellular a- and ß-galactosidase is described using callus cultures of seedlings from the tested plant, roots of 4-6 day old seedlings of Californian poppy germinating on agar plates, and cell suspension cultures cultivated from callus cultures. Synthetic substrates were used for the determination of the intra- and extracellular activities of a- and ß-galactosidase. Many imino- or azasugars are strong glycosidase inhibitors, and some of them show promising chemotherapeutic effects against viral disease and are potential antidiabetic as well as antitumor agents. This fact aroused our interest for a rapid and sensitive assay method of the activity of a- and ß-galactosidases of plant origin. The results presented here show the perspective usefulness of extracellular galactosidases of plant origin in inhibitory and/or biotechnological studies.
A simple, rapid and reproducible procedure for the identification of extracellular Californian po... more 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.
... The same phenomenon (an increase of α-and β-galactosidase activity during storage) was observ... more ... The same phenomenon (an increase of α-and β-galactosidase activity during storage) was observed in the immobilized cells of poppy and ginseng [1921]. The observed increase in the activity on storage remains unclear. ... 5. M. Forster, Enzyme Microbiol. ...
Wageningen Academic Publishers eBooks, Jun 17, 2014
The results of our studies from long-term biomonitoring of polluted environment by means of wild ... more The results of our studies from long-term biomonitoring of polluted environment by means of wild plant species have shown until now fully undiscovered flexibility of nature in resistance to human interference, mainly due to the long-term mutagenesis processes based at auto-regulating and repair mechanisms. The repair ability of nature in the case of the environmental consequences of the war in the Persian Gulf may serve as an example here. Luckily, despite of the biggest ecological catastrophe in modern history, the limits of auto-regulation in the natural environment of the given area were still not crossed. On the contrary, the desert became greener and the sea cleaner than ever before. Adaptation is another mechanisms thanks to which nature turn any violent acts against her to her profit by developing new varieties of life. The fundamental work has shown the successful adaptive response of plant cells even to the most dangerous mutagens. Furthermore, the environment so heavily damaged by human activities that it becomes highly risky for human survival, lower organisms can still adapt – as an example we may mention extremophiles. In this way we may expected the effect of a long-term mutagenesis processes for a creation of new species adapted to the new elements of the environment heavily impacted by human activities.
Cell suspension of Papaver somniferum L. (opium poppy) was permeabilized by Tween 80 and immobili... more Cell suspension of Papaver somniferum L. (opium poppy) was permeabilized by Tween 80 and immobilized by glutaraldehyde. The highest saccharase activity was at pH 4.6 and 50°C. The hydrolysis of the substrate was linear for 5 h reaching 69% of conversion. A very good storage stability was achieved when using dry catalyst, or a solution of 0.15 M NaCl with the addition of chloramphenicol, (1-methyldodecyl)-dimethylamin-4-oxide (ATDNO), chlortetracycline hydrochloride (CLCTC) or by freezing the immobilized cells in 0.15 M NaCl. The cells characterized by high enzyme activity and stability in long-term storage showed convenient technological and physicomechanical properties.
A simple and rapid procedure for the identification and determination of extracellular invertase ... more 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.
Ceská a Slovenská farmacie : casopis Ceské farmaceutické spolecnosti a Slovenské farmaceutické spolecnosti, 2000
A simple, sensitive and reproducible method of detection of extracellular beta-galactosidase was ... more 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.
Chemistry of Natural Compounds - CHEM NAT COMPD, 2002
A simple, rapid, and sensitive procedure for the identification of plant extracellular a- and ß-g... more A simple, rapid, and sensitive procedure for the identification of plant extracellular a- and ß-galactosidase is described using callus cultures of seedlings from the tested plant, roots of 4-6 day old seedlings of Californian poppy germinating on agar plates, and cell suspension cultures cultivated from callus cultures. Synthetic substrates were used for the determination of the intra- and extracellular activities of a- and ß-galactosidase. Many imino- or azasugars are strong glycosidase inhibitors, and some of them show promising chemotherapeutic effects against viral disease and are potential antidiabetic as well as antitumor agents. This fact aroused our interest for a rapid and sensitive assay method of the activity of a- and ß-galactosidases of plant origin. The results presented here show the perspective usefulness of extracellular galactosidases of plant origin in inhibitory and/or biotechnological studies.
A simple, rapid and reproducible procedure for the identification of extracellular Californian po... more 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.
... The same phenomenon (an increase of α-and β-galactosidase activity during storage) was observ... more ... The same phenomenon (an increase of α-and β-galactosidase activity during storage) was observed in the immobilized cells of poppy and ginseng [1921]. The observed increase in the activity on storage remains unclear. ... 5. M. Forster, Enzyme Microbiol. ...
Wageningen Academic Publishers eBooks, Jun 17, 2014
The results of our studies from long-term biomonitoring of polluted environment by means of wild ... more The results of our studies from long-term biomonitoring of polluted environment by means of wild plant species have shown until now fully undiscovered flexibility of nature in resistance to human interference, mainly due to the long-term mutagenesis processes based at auto-regulating and repair mechanisms. The repair ability of nature in the case of the environmental consequences of the war in the Persian Gulf may serve as an example here. Luckily, despite of the biggest ecological catastrophe in modern history, the limits of auto-regulation in the natural environment of the given area were still not crossed. On the contrary, the desert became greener and the sea cleaner than ever before. Adaptation is another mechanisms thanks to which nature turn any violent acts against her to her profit by developing new varieties of life. The fundamental work has shown the successful adaptive response of plant cells even to the most dangerous mutagens. Furthermore, the environment so heavily damaged by human activities that it becomes highly risky for human survival, lower organisms can still adapt – as an example we may mention extremophiles. In this way we may expected the effect of a long-term mutagenesis processes for a creation of new species adapted to the new elements of the environment heavily impacted by human activities.
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