scholarly journals Some characteristics of active and latent monophenolase of mushroom polyphenol oxidase

2013 ◽  
Vol 51 (1-2) ◽  
pp. 33-41 ◽  
Author(s):  
Janusz Czapski
Keyword(s):  
Polyphenol Oxidase ◽  
Quantitative Estimation ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Maximum Velocity ◽  
Electrophoretic Analysis ◽  
Qualitative And Quantitative ◽  
Monophenolase Activity ◽  
Lag Period ◽  
Catalytic Power

Latent form of monophenolase of mushroom polyphenol oxidase (PPO) was activated by 0,1% sodium dodecyl sulfate (SDS). The addition of increasing concentrations of 4-methylcatechol di minished lag period of active and total monophenolase activity, measured using p-cresol with L-proline as a substrate. Changes of lag period were described by equation of one phase exponential decay when concentration of substrate varied from l to 10 mM. Affinity ( 1/K<sub>m</sub>) toward substrate of latent monophenolase was over two times higher than that of the active form, while the maximum velocity (V<sub>max</sub>) was two times lower. The catalytic power (V<sub>max</sub>/K<sub>m</sub>) of both forms of monophenolase were almost equal. Electrophoretic analysis followed by scanning technique of the gels was used. Absorbancy of spots, determined from computer image of isoenzyme bands pattern allowed for qualitative and quantitative estimation of electrophoregrams. Presence of one additional clearly defined slow migrating isoenzyme for SDS activated monophenolase differed in this respect active (2 bands) and total (3 bands) forms of monophenolase.

scholarly journals Some properties of active and latent catechol oxidase of mushroom

2013 ◽  
Vol 52 (1-2) ◽  
pp. 139-147 ◽  
Author(s):  
Janusz Czapski
Keyword(s):  
Kinetic Data ◽  
Sodium Dodecyl ◽  
Electrophoretic Analysis ◽  
Inhibitory Effect ◽  
Catechol Oxidase ◽  
Latent Form ◽  
Salicyl Hydroxamic Acid ◽  
Slow Moving ◽  
Salicyl Hydroxamic ◽  
Catalytic Power

Latent form of mushroom catechol oxidase was activated by O,1% sodium dodecyl sulfate (SDS). Catalytic power of the latent form, calculated from the kinetic parameters was 1,8 times higher than that of active one. Salicyl hydroxamic acid (SHAM) appeared as a powerful inhibitor for both active and latent forms of catechol oxidase. However, in the range of 150-250 μM SHAM the inhibitory effect for active catechol oxidase was significantly higher than that for the latent one. Non-competitive and irreversible characteristics of inhibition of latent and active catechol oxidase was calculated from kinetic data. Electrophoretic analysis followed by scanning of the gels was used. The spots' absorbance was determined from a computer image of the isoenzyme band patterns. It allowed us to estimate gels quantitatively. Presence of one additional clearly defined slow moving isoform of SDS-activated catechol oxidase, differed in the respect of 3 bands for the active and 4 bands for the total.

scholarly journals Qualitative and quantitative determination of phytochemicals from flowers of Spanish Cherry tree

2018 ◽  
Vol 8 (6-s) ◽  
pp. 182-186
Author(s):  
Vineeta Singh ◽  
Anita Rao ◽  
Shipra Pandey ◽  
Vaibhav Sharan Pandey ◽  
Vageshwari Vageshwari ◽  
...  
Keyword(s):  
Quantitative Determination ◽  
Quantitative Estimation ◽  
Standard Procedure ◽  
Tannin Content ◽  
Cherry Tree ◽  
Qualitative And Quantitative ◽  
Standard Curve ◽  
Soxhlet Apparatus ◽  
Percentage Yield

The present enquiry was intended to analyze the phytochemicals qualitatively and quantitatively from flowers of Spanish cherry tree. Flower powder was extracted using polar and nonpolar solvent by soxhlet apparatus. Percentage yield of crude extracts was determined and further the extracts were subjected to analyze the phytochemicals qualitatively and quantitatively by standard procedure. Qualitative analysis showed the absence of alkaloids while presences of tannins, saponins, terpenoids, steroids, glycosides, flavonoids, phenols. Quantitative estimation of phytochemicals was determined using standard curve. Result revealed that the tannin content was 4.3±0.01 (mgTAE /gm), flavanols content was 0.28±0.05 (mgQE/gm). Saponins content was 3.6±0.7 % and terpenoids content was 1.47±0.37 %. A well conducted studies on phytochemicals revealed that they are vital for humans because they provide protection against a variety of ailments. Therefore, the present study is aimed to analyze phytochemicals qualitatively and quantitatively. Keywords: Phytochemicals, Tannins, Saponins, Flavanols, Terpenoids

scholarly journals ON MULTIVARIATE APPROACH TO EVALUATE THE PROTECTION OF AQUIFERS AT DEVELOPMENT OF WESTERN SIBERIA OIL-AND-GAS BEARING AREAS

2015 ◽  
pp. 7-15
Author(s):  
Yu. V. Bespalova
Keyword(s):  
Quantitative Assessment ◽  
Western Siberia ◽  
Oil And Gas ◽  
Quantitative Estimation ◽  
Water Levels ◽  
Multivariate Approach ◽  
Qualitative And Quantitative ◽  
Ground Waters ◽  
Clay Rocks ◽  
Gas Bearing

The qualitative and quantitative assessment of ground waters protectability based on the regional specific charac-teristics of the lithology and thickness of impermeable deposits of the zone of aeration and overlying deposits, a ratio of confined and ground water levels, the soils and clay rocks absorptivity. The article gives the author's vision of ground waters natural protectability. It presents a quantitative estimation by three most well-known methods. Based on the calculations made a map of protectability of fresh ground waters in the Atlym-Novomikhailovsk complex for Nizhnevartovsk district was constructed.

Reversible Sodium Dodecyl Sulfate Activation of Latent Peach Polyphenol Oxidase by Cyclodextrins

2000 ◽  
Vol 379 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Francisco Laveda ◽  
Estrella Núñez-Delicado ◽  
Francisco García-Carmona ◽  
Alvaro Sánchez-Ferrer
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Polyphenol Oxidase ◽  
Sodium Dodecyl ◽  
Dodecyl Sulfate ◽  
Sulfate Activation

scholarly journals The interrelations between high- and low-molecular-weight forms of normal and mutant (Krabbe-disease) galactocerebrosidase

1980 ◽  
Vol 189 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Yoav Ben-Yoseph ◽  
Melinda Hungerford ◽  
Henry L. Nadler
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Krabbe Disease ◽  
Low Molecular Weight ◽  
Molecular Weight Form

Galactocerebrosidase (β-d-galactosyl-N-acylsphingosine galactohydrolase; EC 3.2.1.46) activity of brain and liver preparations from normal individuals and patients with Krabbe disease (globoid-cell leukodystrophy) have been separated by gel filtration into four different molecular-weight forms. The apparent mol.wts. were 760000±34000 and 121000±10000 for the high- and low-molecular-weight forms (peaks I and IV respectively) and 499000±22000 (mean±s.d.) and 256000±12000 for the intermediate forms (peaks II and III respectively). On examination by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the high- and low-molecular-weight forms revealed a single protein band with a similar mobility corresponding to a mol.wt. of about 125000. Antigenic identity was demonstrated between the various molecular-weight forms of the normal and the mutant galactocerebrosidases by using antisera against either the high- or the low-molecular-weight enzymes. The high-molecular-weight form of galactocerebrosidase was found to possess higher specific activity toward natural substrates when compared with the low-molecular-weight form. It is suggested that the high-molecular-weight enzyme is the active form in vivo and an aggregation process that proceeds from a monomer (mol.wt. approx. 125000) to a dimer (mol.wt. approx. 250000) and from the dimer to either a tetramer (mol.wt. approx. 500000) or a hexamer (mol.wt. approx. 750000) takes place in normal as well as in Krabbe-disease tissues.

Preliminary Treatment of Urinary Proteins Improves Electrophoretic Analysis and Immunodetection

1992 ◽  
Vol 38 (6) ◽  
pp. 860-863 ◽  
Author(s):  
J M Verdier ◽  
B Dussol ◽  
P Dupuy ◽  
Y Berland ◽  
J C Dagorn
Keyword(s):  
Protein Pattern ◽  
Protein A ◽  
Sodium Dodecyl ◽  
Complex Mixture ◽  
Electrophoretic Analysis ◽  
Renal Physiology ◽  
Preliminary Treatment ◽  
Urinary Proteins ◽  
Simple Method ◽  
Electrophoretic Migration

Abstract Analysis of urinary protein composition is an important tool in studies on renal physiology and physiopathology. Urine is, however, a complex mixture containing, besides protein, a variety of compounds such as salts, peptides, oligosaccharides, and glycosaminoglycans. Some of these compounds interfere with the electrophoretic migration of protein in sodium dodecyl sulfate-polyacrylamide gels and prevent correct analysis of the protein pattern. We describe a simple method for extracting urinary proteins that considerably improves their electrophoretic migration and subsequent immunodetection. This treatment involves ammonium sulfate fractionations (for precipitating proteins), EDTA (for inhibiting protein aggregation), and HCl hydrolysis (for removing glycosylaminoglycans). Recovery during extraction was found to be almost quantitative for total protein and three representative proteins: albumin, alpha 1-glycoprotein acid, and beta 2-microglobulin.

Two-dimensional electrophoretic analysis of cytosols from human breast tumors: optimal migration conditions.

1984 ◽  
Vol 30 (12) ◽  
pp. 1947-1949 ◽  
Author(s):  
P Motté ◽  
J M Bidart ◽  
J C Delarue ◽  
E Comoy ◽  
P Moingeon ◽  
...  
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Breast Tumors ◽  
Electrophoretic Analysis ◽  
Ph Gradient ◽  
Two Dimensional ◽  
Human Breast ◽  
Cytosolic Proteins ◽  
Optimal Migration ◽  
Dodecyl Sulfate

Abstract In an examination of cytosols from human breast tumors, we performed two-dimensional electrophoretic analysis. Several migration conditions were tried in a search for a homogeneous repartition of cytosolic proteins. The most workable gels were obtained with a 4.40 to 8.05 pH gradient in the first dimension and a homogeneous 125 g/L acrylamide/sodium dodecyl sulfate gel, 1 mm in thickness, in the second dimension. The practicability of this method is discussed.

ON DIFFERENT MOLECULAR EORMS OE PLASMINOGEN ACTIVATOR INHIBITOR

1987 ◽  
Author(s):  
I Carlsson ◽  
J Chmielewska ◽  
B Wiman
Keyword(s):  
Conditioned Medium ◽  
Plasminogen Activator ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Calf Serum ◽  
Gel Filtration ◽  
Plasminogen Activator Inhibitor ◽  
Elution Pattern ◽  
Inactive Form ◽  
Activator Inhibitor

The production of plasminogen activator inhibitor (PAI) by the human cell-lines Hep G2 and HT 1080 have been studied by immunochemical and functional methods. In conditioned medium collected after 2h, the PAI seemed to be almost fully active, but with increasing incubation time the activity was gradually lost, in spite of that the PAI-antigen content increased continously. The active PAI form can be separated from the inactive form by gel-filtration. The inactive form behaves as a low Mr (about 50,000) component in the absence and in the presence of sodium dodecyl-sulphate. In contrast, the active form of PAI behaves as a high Mr (>300,000) compound in the absence of sodium dodecylsulphate but as a low MT compound in its presence. The low M_r inactive PAI has been purified to homogeneity from HT 1080 conditioned medium, collected in the absence of fetal calf serum. This was achieved by chromatography on Concanavalin A-Sepharose, gel-filtration on Sephacryl S-300 and affinity chromatography on insolubilized monoclonal antibodies against PA-inhibitor. On treatment of this form of the inhibitor with 4 mol/L Guanidinium chloride, the activity was regained, but its gel-filtration behaviour was unchanged in the absence of serum/plasma (Mr about 50,000). Addition of plasma or serum prior to the gel-filtration, changed the elution pattern of PAI towards a high Mr form. The reason for this behaviour is not yet fully understood, but the most plausible explanation is the presence of a high Mr PAI-binding protein in plasma/serum. This hypothesis is presently being explored .

Transport and proteolytic processing of rabbit cardiac cathepsin D

1985 ◽  
Vol 248 (1) ◽  
pp. C135-C144
Author(s):  
A. M. Samarel ◽  
A. G. Ferguson ◽  
S. W. Worobec ◽  
M. Lesch
Keyword(s):  
Gel Electrophoresis ◽  
Cathepsin D ◽  
Polyacrylamide Gel Electrophoresis ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Limited Proteolysis ◽  
Proteolytic Processing ◽  
Time Dependent ◽  
Low Density ◽  
Enzyme Maturation

Rabbit cardiac cathepsin D is synthesized as a 53,000-mol wt precursor that undergoes limited proteolysis at an unknown intracellular site to a 48,000-mol wt active form. To examine the site of proteolytic processing, isolated perfused rabbit hearts were fractionated by differential centrifugation 150 or 300 min after pulse labeling with [35S]methionine. Newly synthesized precursor and processed cathepsin D were quantitatively isolated from each fraction by extraction, immunoadsorption, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After 30-min pulse perfusions, all of the 35S-labeled cathepsin D was present as precursor, with the greatest amounts found in low-density subcellular fractions. Proteolytic processing of cathepsin D precursor occurred after chase perfusions that were coincident with the subcellular redistribution of newly synthesized enzyme from sites of synthesis to heavier subcellular structures. Pulse-chase perfusions with chloroquine (10 microM) inhibited precursor proteolytic processing and the time-dependent subcellular redistribution of newly synthesized cathepsin D. The data are consistent with a model for cardiac lysosomal enzyme maturation in which limited proteolytic processing occurs coincident with or soon after the transport of precursors to an acidic intracellular compartment. The results thus suggest that cathepsin D proteolytic processing occurs within cardiac lysosomes.

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