STUDIES ON THE PURIFICATION OF BOVINE PLASMINOGEN (PROFIBRINOLYSIN)

1960 ◽  
Vol 38 (9) ◽  
pp. 1029-1034 ◽  
Author(s):  
Arthur Dalby ◽  
Edmond R. Cole ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Gel Chromatography ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Ph Range

A crude bovine plasminogen product obtained from bovine serum by 30% ammonium sulphate precipitation has been purified 20-fold by means of isoelectric precipitation and calcium phosphate gel chromatography. A threefold purification was achieved by isoelectric precipitation. Plasminogen was precipitated in greatest yield and highest specific activity at 40-fold dilution in the pH range of 5.25–5.50. The conditions under which plasminogen is eluted from calcium phosphate gel columns have been investigated. Plasminogen fractions possessing specific activity seven times that of the isoelectric-precipitated materials have been obtained by elution with phosphate buffers over the range of 0.05 to 0.1 M, pH 6.8.

STUDIES ON THE PURIFICATION OF BOVINE PLASMINOGEN (PROFIBRINOLYSIN)

1960 ◽  
Vol 38 (1) ◽  
pp. 1029-1034 ◽  
Author(s):  
Arthur Dalby ◽  
Edmond R. Cole ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Gel Chromatography ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Ph Range

A crude bovine plasminogen product obtained from bovine serum by 30% ammonium sulphate precipitation has been purified 20-fold by means of isoelectric precipitation and calcium phosphate gel chromatography. A threefold purification was achieved by isoelectric precipitation. Plasminogen was precipitated in greatest yield and highest specific activity at 40-fold dilution in the pH range of 5.25–5.50. The conditions under which plasminogen is eluted from calcium phosphate gel columns have been investigated. Plasminogen fractions possessing specific activity seven times that of the isoelectric-precipitated materials have been obtained by elution with phosphate buffers over the range of 0.05 to 0.1 M, pH 6.8.

PARTIAL PURIFICATION OF BOVINE ANTIPLASMIN (ANTIFIBRINOLYSIN)

1960 ◽  
Vol 38 (9) ◽  
pp. 1021-1027
Author(s):  
Edmond R. Cole ◽  
Arthur Dalby ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Large Diameter ◽  
Small Diameter ◽  
Partial Purification ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Column Eluate

Bovine antiplasmin (antifibrinolysin) solutions with activities of 12–19 units/mg nitrogen have been obtained from bovine serum by 60% ammonium sulphate precipitation and isoelectric precipitation at pH 5.0 representing approximately a 6-fold purification of antiplasmin from serum. An additional 6.4-fold purification of antiplasmin was obtained by chromatography on a large diameter calcium phosphate gel column. Dialysis and lyophilization of the column eluate caused a 30% loss in activity. The dry antiplasmin was 25 times more potent than serum protein and was stable for 12 months when stored in a desiccator over P2O5 at −10°. Antiplasmin solutions with a specific activity twice that obtained with the large column could be eluted from a small diameter calcium phosphate column, but the activity decreased too rapidly to permit preparation of a lyophilized product.

PARTIAL PURIFICATION OF BOVINE ANTIPLASMIN (ANTIFIBRINOLYSIN)

1960 ◽  
Vol 38 (1) ◽  
pp. 1021-1027
Author(s):  
Edmond R. Cole ◽  
Arthur Dalby ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Large Diameter ◽  
Small Diameter ◽  
Partial Purification ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Column Eluate

Bovine antiplasmin (antifibrinolysin) solutions with activities of 12–19 units/mg nitrogen have been obtained from bovine serum by 60% ammonium sulphate precipitation and isoelectric precipitation at pH 5.0 representing approximately a 6-fold purification of antiplasmin from serum. An additional 6.4-fold purification of antiplasmin was obtained by chromatography on a large diameter calcium phosphate gel column. Dialysis and lyophilization of the column eluate caused a 30% loss in activity. The dry antiplasmin was 25 times more potent than serum protein and was stable for 12 months when stored in a desiccator over P2O5 at −10°. Antiplasmin solutions with a specific activity twice that obtained with the large column could be eluted from a small diameter calcium phosphate column, but the activity decreased too rapidly to permit preparation of a lyophilized product.

scholarly journals Biobleaching of Industrial Important Dyes with Peroxidase Partially Purified from Garlic

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Akudo Chigozirim Osuji ◽  
Sabinus Oscar O. Eze ◽  
Emmanuel Emeka Osayi ◽  
Ferdinand Chiemeka Chilaka
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Low Cost ◽  
Gel Filtration ◽  
Enzyme Concentration ◽  
Gel Filtration Chromatography ◽  
Vat Dyes ◽  
Ammonium Sulphate Precipitation ◽  
Ph Range ◽  
Better Than

An acidic peroxidase was extracted from garlic (Allium sativum) and was partially purified threefold by ammonium sulphate precipitation, dialysis, and gel filtration chromatography using sephadex G-200. The specific activity of the enzyme increased from 4.89 U/mg after ammonium sulphate precipitation to 25.26 U/mg after gel filtration chromatography. The optimum temperature and pH of the enzyme were 50°C and 5.0, respectively. The Km andVmaxfor H2O2and o-dianisidine were 0.026 mM and 0.8 U/min, and 25 mM and 0.75 U/min, respectively. Peroxidase from garlic was effective in decolourizing Vat Yellow 2, Vat Orange 11, and Vat Black 27 better than Vat Green 9 dye. For all the parameters monitored, the decolourization was more effective at a pH range, temperature, H2O2concentration, and enzyme concentration of 4.5–5.0, 50°C, 0.6 mM, and 0.20 U/mL, respectively. The observed properties of the enzyme together with its low cost of extraction (from local sources) show the potential of this enzyme for practical application in industrial wastewater treatment especially with hydrogen peroxide. These Vat dyes also exhibited potentials of acting as peroxidase inhibitors at alkaline pH range.

Anreicherung und Charakterisierung einer „Transhydrogenase-Aktivität“ und der 17 β-Hydroxysteroid-Oxidoreduktase aus der Cytoplasma-Fraktion der Leber von weiblichen Ratten / The Enrichment and Characterisation of Cytoplasmatic “Transhydrogenase-activity” and 17 β-hydroxysteroid-oxidoreductase from Rat Liver

1972 ◽  
Vol 27 (8) ◽  
pp. 981-988
Author(s):  
Kunhard Pollow ◽  
Barbara Pollow
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulphate ◽  
Rat Liver ◽  
Isoelectric Focusing ◽  
Isoelectric Point ◽  
Column Chromatography ◽  
Enzymatic Activities ◽  
Kinetic Constants ◽  
Gel Chromatography ◽  
Ammonium Sulphate Precipitation

The cytoplasmatic fraction of rat liver contains both 17 β-hydroxysteroid-oxidoreductase and a “transhydrogenase-activity”, which catalyses the transfer of hydrogen from the 17 β-position of estradiol-17 β to the 17-position of 4-androstene-3,17-dione. The 17 β-hydroxysteroid-oxidoreductase was purified 718-fold and the “transhydrogenase-activity” 264-fold by ammonium sulphate precipitation, gel chromatography with Sephadex G-200, column chromatography on DEAE-Sephadex and isoelectric focusing. The two enzymic activities could not be separated. The characteristics of the two enzymatic activities give some evidence that the “transhydrogenase-activity” is identical with the already known 17 β-hydroxysteroid-oxidoreductase.Isoelectric focusing of the chromatographycally enriched 17 β-enzyme gave an isoelectric point at 5,2. The 17 β-enzyme has a molecular weight of 62 — 65 000 as determined by mobility on Sephadex G-200 superfine.The kinetic constants for both the 17 β-enzyme and the “transhydrogenase-activity” were determined.

Fermentative Production, Purification and Characterization of Nisin

2008 ◽  
Vol 4 (5) ◽  
Author(s):  
Swapnali S Gujarathi ◽  
Sandip B. Bankar ◽  
Laxmi A. Ananthanarayan
Keyword(s):  
Ammonium Sulphate ◽  
Hydrophobic Interaction ◽  
Orthogonal Array ◽  
Gel Filtration ◽  
Statistical Design ◽  
Temperature Stability ◽  
Gel Chromatography ◽  
Purification And Characterization ◽  
Ammonium Sulphate Precipitation

Bacteriocins are bactericidal or bacteriostatic in action and active against closely related species. Nisin is the bacteriocin produced by the lactic acid bacteria (LAB). Nisin is a small (3353 Da), cationic, hydrophobic, and 34-amino acid peptide. It is used in products such as pasteurized processed cheese, salad dressing, and liquid whole eggs to inhibit the growth of Gram-positive microorganisms including Listeria monocytogenes. The objective of the present work was to study production, purification and characterization of nisin. The production of nisin was carried out using Lactococcus lactis subsp lactis MTCC 440 by one-factor-at-a-time method and statistical design (Orthogonal array). Purification was carried out using ammonium sulphate precipitation followed by hydrophobic interaction and gel chromatography. Characterization was done for pH and temperature stability. The activity of nisin after one factor at a time optimization was found to be 5120 AU/ml. The activity of the nisin increased to 6800 AU/ml after optimization by Orthogonal Array design. Ammonium sulphate precipitation gives good yield of nisin with 60 to 80% saturation with 2.52 fold purity. The overall purification by hydrophobic interaction and gel filtration chromatography was 10.87 fold with 50.84% yield and 8.8 fold with 49.65% yield as compared to crude broth respectively.

scholarly journals Bromelain Activity of Waste Parts of Two Pineapple Varieties

2018 ◽  
Vol 2 ◽  
pp. 21-28 ◽  
Author(s):  
Edmund Ofosu Benefo ◽  
Isaac Williams Ofosu
Keyword(s):  
Enzyme Activity ◽  
Ammonium Sulphate ◽  
Crude Extract ◽  
Specific Activity ◽  
High Demand ◽  
Digestion Method ◽  
Waste Products ◽  
Food Industries ◽  
Ethanol Precipitation ◽  
Ammonium Sulphate Precipitation

Bromelain, a protease found in pineapples, is of high demand in the pharmaceutical, cosmetic and food industries. Along the pineapple processing chain, waste products such as peels, crowns, stems and cores result. These parts are usually discarded, though they contain significant amounts of the enzyme bromelain. This study sought to determine the bromelain activity of the crowns and peels of two pineapple varieties grown in Ghana;MD2andSugarloaf. The crude extract was obtained by homogenising the peels and crowns in a cold phosphate buffer and centrifuging at 3000 rpm for 15 min. Ethanol and ammonium sulphate precipitation were carried out on the extract between 30% – 80% precipitation levels. The enzyme activity was determined using the casein digestion method. Results showed that bromelain was precipitated mainly in the 30% – 60% precipitation range.Sugarloafcrowns yielded the highest enzyme activity of 20.82 U/ml and a specific activity of 194.58 U/mg at the 40% ammonium sulphate precipitation level. This was followed by theSugarloafpeels with an enzyme activity of 19.98 U/ml at 50% ethanol precipitation level. Ethanol precipitation resulted in fractions with lower bromelain activity. Enzyme activity was higher in theSugarloafvariety and also in the crowns of both varieties. The two pineapple varieties have significant levels of bromelain activity and could be exploited for commercialisation.

THE DEVELOPMENT OF A RADIOIMMUNOASSAY FOR OXYTOCIN: RADIO-IODINATION, ANTIBODY PRODUCTION AND SEPARATION TECHNIQUES

1970 ◽  
Vol 46 (2) ◽  
pp. 269-278 ◽  
Author(s):  
T. CHARD ◽  
M. J. KITAU ◽  
J. LANDON
Keyword(s):  
Lymph Node ◽  
Human Plasma ◽  
Rapid Method ◽  
Ammonium Sulphate ◽  
Antibody Production ◽  
Specific Activity ◽  
High Specific Activity ◽  
Separation Techniques ◽  
Ammonium Sulphate Precipitation

SUMMARY A simple and rapid method is described for labelling oxytocin with 131I at a high specific activity. This method is compared with those of previous workers. A satisfactory antiserum has been raised by direct intra-lymph node injection of oxytocin adsorbed to carbon microparticles. A number of methods for separating antibody-bound from free oxytocin are described, and reasons given for preferring a procedure using ammonium sulphate precipitation. These data form the basis for developing a radioimmunoassay intended for the determination of oxytocin in human plasma.

Purification of a Fibrinolytic Enzyme from Bacillus Sp. Isolated from Budu

2012 ◽  
Vol 59 (1) ◽  
Author(s):  
Nurulhanis Ahmad Sanusi ◽  
Haryati Jamaluddin
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Culture Broth ◽  
Fibrinolytic Enzyme ◽  
Total Protein Content ◽  
Bacillus Sp ◽  
Ammonium Sulphate Precipitation ◽  
Sds Page

Bacillus sp. strain B1 producing wild type fibrinolytic enzyme was isolated from Budu. The fibrinolytic enzyme was collected from the supernatant of Bacillus sp. strain B1 culture broth and purified to electrophoretic homogeneity through a combination of various purification schemes, which include ammonium sulphate precipitation, followed by anion exchange chromatography using DEAE–Sepharose Fast Flow and gel filtration chromatography on Sephadex G–75 column. During ammonium sulphate precipitation screening, it was observed that the crude enzyme from Bacillus sp. strain B1 precipitated at 40% and 50% of ammonium sulphate saturation respectively. The fibrinolytic enzyme was purified 58.5–fold with a final yield of 0.51%. The specific activity was determined to be 1.17 Units/mg using plasmin as standard and the final total protein content was 8.58 mg/ml. After the successive purification steps, the estimated molecular mass of fibrinolytic enzymes from strain B1 was estimated via sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE). SDS–PAGE analysis showed a single band at 45 kDa corresponding to the purified fraction with fibrinolytic activity.

scholarly journals Purification of soluble arylsulphatases from ox brain

1965 ◽  
Vol 97 (2) ◽  
pp. 360-364 ◽  
Author(s):  
W Bleszynski ◽  
LM Dzialoszynski
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Ammonium Sulphate Precipitation ◽  
Optimum Ph ◽  
Arylsulphatase A

1. Two soluble arylsulphatases (A and B) have been extracted from ox brain by a modified Albers autolysis method and purified by acetone and ammonium sulphate precipitation and dialysis. 2. A 1600-fold purification was achieved with arylsulphatase A and 320-fold purification with arylsulphatase B. 3. The specific activity of arylsulphatase A was 266000 4-nitrocatechol units/mg. of protein N, and that of arylsulphatase B was 64600units/mg. of protein N. 4. Arylsulphatase A seems to be electrophoretically homogeneous. 5. With 3mm-dipotassium 2-hydroxy-5-nitrophenyl sulphate as substrate the optimum pH for the activity of arylsulphatase A was 4.7, and for arylsulphatase B it was 6.1 with a 60mm solution of the same substrate.

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