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.

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.

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.

PURIFICATION AND CHARACTERIZATION OFESTERASE ENZYME ISOLATED FROM CONTAMINATED SOIL SAMPLE

2019 ◽  
pp. 1-3
Author(s):  
S. Chithra ◽  
K. Silambarasi ◽  
I. Yasmin
Keyword(s):  
Soil Sample ◽  
Ammonium Sulphate ◽  
Contaminated Soil ◽  
Specific Activity ◽  
Nitrogen Sources ◽  
Partial Purification ◽  
Carbon And Nitrogen ◽  
Crude Enzyme Preparation ◽  
Ammonium Sulphate Precipitation ◽  
Highly Active

Isolation and partial purification of esterase from contaminated soil sample was the main aim of this study. The production medium for organism was optimized by using different pH,Temperature,Carbon and Nitrogen sources.The esterase enzyme was highly active and stable from pH 5.0 to 9.0 with an optimum at pH 9.Its optimum temperature was 35°C. The best carbon and nitrogen sources were mannitol and yeast extract. Esterase was partially purified by ammonium sulphate precipitation,dialysis.The specific activity of partially purified esterase obtained from ammonium sulphate fractionation is found to be 8.6485U/mg and the fractionation is 5.4 fold purified from the crude enzyme preparation yielding 17.5U/mg from the crude protein. This result showed that Bacillus subtilis under study is a good producer of extra cellular esterase,which can be beneficial for industries

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 New Lipase for Biodiesel Production: Partial Purification and Characterization of LipSB 25-4

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Aysel Ugur ◽  
Nurdan Sarac ◽  
Rukiye Boran ◽  
Berk Ayaz ◽  
Ozgur Ceylan ◽  
...  
Keyword(s):  
Specific Activity ◽  
Gel Filtration ◽  
Nitrogen Sources ◽  
Biodiesel Production ◽  
Partial Purification ◽  
Period Effect ◽  
Carbon And Nitrogen ◽  
Ammonium Sulphate Precipitation ◽  
Thermophilic Conditions

The lipolytic activities of 300 Streptomyces isolates were determined in Tributyrin and Rhodamine-B Agar. Lipase activities were also measured with p-nitrophenyl palmitate (p-NPP) as a substrate. The strain of Streptomyces bambergiensis OC 25-4 used in this study was selected among 300 strains of Streptomyces from MUCC as the best lipase producer. The incubation conditions were optimized and the inoculum amount, incubation period, effect of carbon and nitrogen sources, and rates of MgSO4 and CaCO3 were investigated. LipSB 25-4 (the lipase produced by S. bambergiensis OC 25-4 strain) was partially purified with ammonium sulphate precipitation, dialysis, and gel filtration chromatography 2.73-fold and with 92.12 U/mg specific activity. The optimal pH and temperature for LipSB 25-4 were determined as 8.0 and 50°C, respectively. The lipase has high stability in all pH and temperature values used in this study. While LipSB 25-4 was slightly activated in the presence of β-mercaptoethanol, it was slightly reduced by PMSF. The enzyme conserved approximately 75% of its activity at the end of 60 h, in the presence of methanol and ethanol. Since LipSB 25-4 displays high activity in the thermophilic conditions and stability in the presence of organic solvents, this lipase can catalyse the biodiesel production from olive oil by the transesterification reactions.

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.

scholarly journals PEMURNIAN PARSIAL DAN KRISTALISASI PAPAIN DARI GETAH Carica papaya

2019 ◽  
Vol 4 (2) ◽  
pp. 152
Author(s):  
Dwi Putri Mashfufatur Rohmah ◽  
Sofijan Hadi ◽  
Afaf Baktir
Keyword(s):  
Ammonium Sulphate ◽  
Carica Papaya ◽  
Specific Activity ◽  
Total Activity ◽  
Partial Purification ◽  
Purification Factor ◽  
Papaya Latex ◽  
Carica Papaya Latex

AbstractThis research has done partial purification by fractionation and optimization crystallization of papain from Carica papaya latex with the addition of ammonium sulphate. The enhancement of purification factor was determined by its specific activity. The fractionation results show that papain fraction of Carica papaya can be obtained by adding 40-80% saturated ammonium sulphate, with the highest specific activity, i.e. 2,0819 U/µg and the factor purification increase of 6,27 fold than papain extract. Meanwhile, the highest total activity, i.e. 10,7780 U of papain crystals can be obtained by presipitant agent addition of ammonium sulphate in the level / concentration 80% saturated at 15 °C. Microscopycally papain crystals profile in this condition have cube and tetragonal shape.Key words: crystallization, fractionation, ammonium sulphate, papain

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