Characterization of the Esterases of Feline Serum

1974 ◽  
Vol 52 (11) ◽  
pp. 1073-1078 ◽  
Author(s):  
D. J. Ecobichon
Keyword(s):  
Close Association ◽  
Gel Filtration ◽  
Spectrophotometric Analysis ◽  
Specific Substrate ◽  
Starch Gel Electrophoresis ◽  
Kinetic Characteristics ◽  
Starch Gel ◽  
Low Levels ◽  
Optimal Ph

Various techniques, including electrophoresis, gel filtration, titrimetric, and spectrophotometric analysis with specific and nonspecific substrates and inhibitors, were used to characterize the serum esterases of male and female cats. Starch-gel electrophoresis yielded five bands of activity: three butyrylcholinesterase bands and two bands of nonspecific carboxylesterase activity. Gel filtration on Sephadex G-200 yielded two distinct peaks of activity, one containing the cholinesterase bands and the other, the carboxylesterase activity. Kinetic characteristics determined for feline serum esterases included: (1) optimal pH; (2) optimal substrates for cholinesterase and carboxylesterase activities; (3) Km values for the choline, α-naphthyl, and p-nitrophenyl esters. With the organophosphate paraoxon as a specific substrate for arylesterase activity, low levels of this enzyme could be detected in close association with the carboxylesterase.

Characterization of the esterases of canine serum

1970 ◽  
Vol 48 (12) ◽  
pp. 1359-1367 ◽  
Author(s):  
D. J. Ecobichon
Keyword(s):  
Close Association ◽  
Gel Filtration ◽  
Spectrophotometric Analysis ◽  
Specific Substrate ◽  
Starch Gel ◽  
Males And Females ◽  
Canine Serum ◽  
Hydrolysis Of ◽  
Naphthyl Acetate

Various techniques including electrophoresis, gel filtration, and titrimetric and spectrophotometric analysis with specific and nonspecific substrates and inhibitors were used to characterize the serum esterases of male and female dogs. Electrophoresis in starch gel yielded 10 distinct bands of activity: four butyrylcholinesterase bands, an esterolytically active albumin, and five bands of aliesterase and/or arylesterase activity. Gel filtration on Sephadex G-200 yielded two distinct peaks of activity, one containing the cholinesterase bands, and the second peak containing both arylesterase and aliesterase activity in close association with the serum albumin. Kinetic characteristics determined for the canine serum esterases included (1) optimal pH; (2) Km values for esters of choline, α-naphthol, and p-nitrophenol; and (3) average rates of hydrolysis of α-naphthyl acetate, butyrylcholine iodide, p-nitrophenyl acetate, and E600 by the sera of males and females. The sensitivity of the serum esterases to inhibition by various cations, and specific and nonspecific inhibitors was investigated. The organophosphate, E600, may be a highly specific substrate for the detection and quantification of arylesterase in a mixture of nonspecific carboxylesterases.

Purification and characterization of the polyphenoloxidase (PPO) isolated from Solenocera crassicornis (H. Milne Edwards, 1837)

Crustaceana ◽  
2015 ◽  
Vol 88 (3) ◽  
pp. 259-272 ◽  
Author(s):  
X. W. Xiang ◽  
Y. F. Zhou ◽  
Y. B. Hao ◽  
H. C. Yang ◽  
H. W. Ma ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Citric Acid ◽  
Ethylenediaminetetraacetic Acid ◽  
Gel Filtration ◽  
Catechol Oxidase ◽  
Specific Substrate ◽  
Purification And Characterization ◽  
Sds Page ◽  
Optimal Ph

Polyphenoloxidase (PPO) from Solenocera crassicornis (H. Milne Edwards, 1837) was partially purified through ion-exchange and gel-filtration chromatography, and its enzymatic properties were also characterized using l-dihydroxyphenylalanine (L-DOPA) as the specific substrate. The molecular mass of PPO in SDS-PAGE is about 66.5 kDa, and the PPO molecule isolated by HPLC, is 133 kDa, implying it is a homodimeric protein. The optimal pH and temperature of PPO activity was 6.5 and 40°C, and the value was 3.80 and 8.12 on L-DOPA and on catechol, respectively. It was found that the purified PPO could only catalyse o-diphenol substrates, inferring that it belongs to the catechol-oxidase family. The PPO was very sensitive to phenylthiourea, ascorbic acid, citric acid and cysteine. The PPO activity was strongly inhibited by ethylenediaminetetraacetic acid (EDTA), Cu2+ and Zn2+, indicating that Solenocera crassicornis PPO is most probably a copper-containing metalloenzyme.

scholarly journals A chromatographic preparation of ox growth hormone

1966 ◽  
Vol 100 (3) ◽  
pp. 593-600 ◽  
Author(s):  
M Wallis ◽  
HBF Dixon
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Gel Electrophoresis ◽  
Anterior Pituitary ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Highly Active ◽  
Cross Linkage ◽  
Starch Gel

1. A method is described for the chromatographic preparation of ox growth hormone. It involves chromatography of an extract of anterior pituitary lobes on DEAE-cellulose, followed by rechromatography on a dextran gel of low cross-linkage (Sephadex G-100). 2. The product is highly active in growth-hormone assays, and is obtained in good yield. It was homogeneous by several criteria, but showed some heterogeneity on starch-gel electrophoresis. 3. The molecular weight of the hormone was estimated from its behaviour on gel-filtration columns under various conditions. Evidence that the hormone may dissociate into sub-units under some conditions is presented.

scholarly journals Haemoglobin polymorphism in selected farm animals: A review

2014 ◽  
Vol 30 (3) ◽  
pp. 377-390 ◽  
Author(s):  
S.S.A. Egena ◽  
R.O. Alao
Keyword(s):  
Genetic Improvement ◽  
Raw Materials ◽  
Farm Animals ◽  
Starch Gel Electrophoresis ◽  
Gene Frequencies ◽  
Starch Gel ◽  
And Performance ◽  
Polypeptide Chains ◽  
Degree Of Similarity

Biochemical diversity or polymorphism is the occurrence of varieties attributed to biochemical differences which are under genetic control. It has created a leeway for the genetic improvement of farm animals. This is because it can be used as a useful tool for the characterization of livestock breeds and population. This way, the degree of similarity or differences within and between breeds can be ascertained and this differences or similarity are important raw materials for genetic improvement of animals. Data obtained on gene frequencies and genotypes through polymorphism study makes it not only possible to compare the gene stocks of animals, the possible effects of the genes on reproductive and performance traits, but also study genetic variability under different environmental conditions of selection. This study was carried out to review haemoglobin (Hb) polymorphism in selected farm animals with the view of finding out the type of polymorphism observed by starch gel electrophoresis due to variation in the amino acid sequence in the polypeptide chains of Hb. The review showed clearly that there is a gene-controlled diversity in the different farm animals considered. This could serve as a reference point for future studies earmarked for the improvement of the animals possibly via marker-assisted selection.

Isolation and characterization of Schizosaccharomyces pombe mutants with defective NAD-dependent malic enzyme

1986 ◽  
Vol 32 (6) ◽  
pp. 481-486 ◽  
Author(s):  
C. Osothsilp ◽  
R. E. Subden
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Malic Enzyme ◽  
Pool Analysis ◽  
Starch Gel Electrophoresis ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Starch Gel ◽  
Colony Color ◽  
Color Indicator

To obtain NAD-dependent malic enzyme mutants of Schizosaccharomyces pombe, a colony color indicator screening system was developed. Mutants defective for malic acid utilization (mau mutants) are yellow, while wild-type colonies are blue on the defined bromcresol green based indicator medium. NAD-dependent malic enzyme mutants were distinguished from other mau mutants by subsequent, starch gel electrophoresis, spectrophotometry, complementation tests, and intermediate pool analysis with cell-free extracts.

FAMILIAL GOITRE WITH ABSENCE OF THYROGLOBULIN AND SYNTHESIS OF THYROID HORMONES FROM THYROIDAL ALBUMIN

1974 ◽  
Vol 60 (3) ◽  
pp. 389-397 ◽  
Author(s):  
K. B. DESAI ◽  
M. N. MEHTA ◽  
M. C. PATEL ◽  
S. M. SHARMA ◽  
L. RAMANNA ◽  
...  
Keyword(s):  
Thyroid Hormones ◽  
Radioactive Iodine ◽  
Gradient Centrifugation ◽  
Gel Filtration ◽  
Sucrose Density Gradient ◽  
Starch Gel Electrophoresis ◽  
Sucrose Density Gradient Centrifugation ◽  
Salting Out ◽  
Immunological Tests ◽  
Starch Gel

SUMMARY Two siblings, a brother (H. B.) and a sister (R. B.) with long standing goitres were investigated. Radioactive iodine uptake by the thyroid was increased and a significant portion of the plasma radioactive iodine was not extractable with butanol. Chromatography of butanol extracts of serum after radioactive iodine administration showed distinct peaks of triiodothyronine and thyroxine. Microscopic examination of the surgical specimens of the goitres showed Hürthle cell carcinoma with follicles devoid of colloid in both specimens. Sucrose density gradient centrifugation, gel filtration on Sephadex G-200, salting out procedures, starch gel electrophoresis and immunological tests of the supernatant soluble fraction of thyroid homogenates showed a lack of thyroglobulin. Further fractionation of the soluble proteins showed that albumin was apparently involved in the synthesis of thyroid hormones in the absence of thyroglobulin.

Cytoplasmic Carboxylesterases of Human and Domestic Animal Liver: Aggregation, Dissociation and Molecular Weight Estimation

1972 ◽  
Vol 50 (1) ◽  
pp. 9-15 ◽  
Author(s):  
D. J. Ecobichon
Keyword(s):  
Molecular Weight ◽  
Human Liver ◽  
Gel Filtration ◽  
Domestic Animal ◽  
Starch Gel Electrophoresis ◽  
Weight Estimation ◽  
Animal Liver ◽  
Starch Gel ◽  
Species Specific ◽  
Two Peaks

The cytoplasmic carboxylesterases of bovine, ovine, equine and human liver were fractionated by starch gel electrophoresis and by gel filtration on Sephadex. While species-specific, heterogeneous bands were observed in starch gel, the esterases of the bovine, ovine and equine liver were eluted from Sephadex G-100 as single peaks of activity, each with a characteristic elution volume. Gel filtration of human liver extracts yielded two peaks of activity, one containing electrophoretically slow esterases, the other electrophoretically fast esterases. Extracted equine and human hepatic carboxylesterases aggregated readily on storage or concentration, forming larger units which could be dissociated by a combination of acidic pH and high salt concentration. Molecular weight estimates of the hepatic esterases by gel filtration on Sephadex G-100 and G-200 yielded values of 65 000 for ovine, 55 000 for bovine, 96 000 and 70 000 for equine variants and 180 000 and 65 000 for human variants. The observations suggested that the cytoplasmic enzymes in relatively crude hepatic extracts had a lower molecular weight than those in concentrated or partially purified preparations which formed stable dimers or trimers.

Biochemical characterization of Tunisian grapevine varieties

OENO One ◽  
1998 ◽  
Vol 32 (1) ◽  
pp. 17
Author(s):  
Ferjani Ben Abdallah ◽  
Farhat Chibani ◽  
Asma Fnayou ◽  
Abdelwahed Ghorbel ◽  
Jean-Michel Boursiquot
Keyword(s):  
Biochemical Characterization ◽  
Biochemical Study ◽  
Starch Gel Electrophoresis ◽  
Banding Patterns ◽  
Enzyme Systems ◽  
Mother Plants ◽  
Starch Gel ◽  
Biochemical Identification ◽  
Berry Colour

<p style="text-align: justify;">61 tunisian autochton grapevine varieties have been collected for biochemical identification. Isozymes analysis with starch gel electrophoresis technique was used to confirn or to cancel random denominations awarded to the majority of these local varieties. In our conditions, concentrated plant extracts were obtained from vigorous donnant canes newly cut off from selected mother plants during automn. These allowed us to dispose of rigorously interpretable isozyme banding patterns of GPI and PGM systems and to overcome difficulties often related to the use of PGM system. The study of GPII and PGM enzyme systems allowed us to classify the autochton accessions into 16 different groups from which 5 groups containing only 2 or 3 varieties.</p><p style="text-align: justify;">On the other hand, the study of AAT and peroxydase enzyme systems has shown stable and legible isozyme banding patterns allowing to discriminate between equivalent accessions such as Sakasly and Kahli (two black local vines very similar), 3 varieties of Bidh Hamem (Bidh Hamem, Bidh Hamem Rafraf and Bidh Hamem Sfax), and 2 varieties of Bezzoul Kelba Bidha (Sfax and Gabes). In addition, certain varieties having for longtime the same denominations were characterized. A case of point the 4 varieties Khalt meaning mixture (Bouchemma, Abiedh, Mdaouer and Souche 1) and the 3 varieties of Arich (Ahmar, Dressée, and Jerba) were proved to be completely different from each other. In the same way, Bezzoul Khadem has been differed from Hemri variety. The complementary use of berry colour allowed to discriminate between Saouadi, Khdhiri and Jebbi varieties and to subdivise the remainig groups into sub-groups.</p><p style="text-align: justify;">The study of GPI, PGM, AAT and peroxydase isozyme banding patterns in combination with berry colour has led to establish a classification of the 61 autochton varieties into 37 groups including 26 varieties definitely differentiated through the results of this biochemical study.</p>

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