Table 2. Ammonium sulfate saturation table using solid ammonium sulfate

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.tab2
Keyword(s):  
Ammonium Sulfate ◽  
Ammonium Sulfate Saturation

scholarly journals Partial purification of thrombopoietin from the plasma of thrombocytopenic rabbits

Blood ◽  
1979 ◽  
Vol 54 (2) ◽  
pp. 377-388 ◽  
Author(s):  
BL Evatt ◽  
J Levin ◽  
KM Algazy
Keyword(s):  
Ammonium Sulfate ◽  
Phosphate Buffer ◽  
Carboxymethyl Cellulose ◽  
Room Temperature ◽  
Deae Cellulose ◽  
Partial Purification ◽  
Elution Volume ◽  
Ammonium Sulfate Saturation ◽  
Citrate Phosphate ◽  
Citrate Phosphate Buffer

Abstract Partially purified thrombopoiesis-stimulating activity was prepared from the plasma of thrombocytopenic rabbits using ammonium sulfate precipitation and DEAE cellulose, Sephadex, and carboxymethyl cellulose chromatography. The protein fraction precipitated by an ammonium sulfate saturation of 60%-80%, previously shown to contain thrombopoiesis-stimulating activity, was used as starting material. Column chromatography was carried out at room temperature at pH 5.6. Under these conditions, thrombopoiesis-stimulating activity (thrombopoietin) was retained by DEAE cellulose (0/03 M citrate- phosphate buffer) and carboxymethyl cellulose (0/003 M citrate- phosphate buffer), and eluted with 0.4 M NaCl. Thrombopoietin was retarded by Sephadex G-100; the ratio of the elution volume to the void volume was 1.32:1. Immunoelectrophoretic analysis of partially purified thrombopoietin indicated that following removal of most of the albumin by DEAE chromatography, only proteins with the mobilities of beta- globulins and albumin and traces of other anodally migrating proteins were detectable in the fractions that contained thrombopoiesis- stimulating activity. Thrombopoietin was not dialyzable and was stable from at least pH 5.6 to 7.5. It was approximately 1000-fold purified following sequential chromatography with DEAE and carboxymethyl cellulose. Although the three fractions described reproducibly stimulated thrombopoiesis, as measured by increased levels of selenomethionine-75Se (75SeM) in the circulating platelets, platelet counts did not increase.

Partial Purification and Characterization of Polyphenol Oxidase from the Wild Edible Mushroom Lepiota Procera Using Three-Phase Partitioning

2018 ◽  
Vol 14 (9-10) ◽  
Author(s):  
Neslihan Saki ◽  
Mustafa Akin ◽  
Esma H. Alici ◽  
Gulnur Arabaci
Keyword(s):  
Ammonium Sulfate ◽  
Polyphenol Oxidase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Edible Mushroom ◽  
Bottom Phase ◽  
Phase Partitioning ◽  
Three Phase ◽  
Ammonium Sulfate Saturation ◽  
Wild Edible Mushroom

AbstractPolyphenol oxidase (PPO) from the wild edible mushroom Lepiota procera is partially purified and biochemically characterized using three-phase partitioning (TPP), which is an easily applied and effective method. This method includes ammonium sulfate saturation at different concentrations, t-butanol addition (1:1; 1:5), and adjustment of pH. Optimum purification parameters with 20% ammonium sulfate saturation and 1:1 t-butanol conditions led to the highest activity at bottom phase with 8.4 fold purification. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed the molecular weight of the purified enzyme to be 35 kDa. The partially purified PPO presented a high level of activity with L-DOPA (Michaelis-Menten constant, Km – 0.12 mM), followed by caffeic acid (0.27 mM) and 4-methylcatechol (0.46 mM) and it is classified as a catecholase type of PPO. The enzyme had peak activity at a temperature of 40 °C and a pH value of 7.0. These results demonstrate a new enzyme source and easy purification method, useful for various industrial applications.

scholarly journals Identification and Purification of Cholera Like Toxin from Environmental Isolate of Vibrio cholerae

2010 ◽  
Vol 7 (1) ◽  
pp. 415-423
Author(s):  
Baghdad Science Journal
Keyword(s):  
Vibrio Cholerae ◽  
Ammonium Sulfate ◽  
Water Samples ◽  
Specific Activity ◽  
Gel Filtration ◽  
Environmental Isolate ◽  
Back Extraction ◽  
Isolation Techniques ◽  
Purification Scheme ◽  
Ammonium Sulfate Saturation

The presence and prevalence of V. cholerae were investigated in forty five water samples collected from different locations of Tiger River/ Baghdad city. Twenty one isolates were isolated by adopting a simple isolation techniques. The final identification revealed that only three isolates were confirmed as V. cholerae. They were named 1J, 1R and Dial 131 which are all serogrouped as non-O1. Toxin Coregulated Pili (TCP) and heat labile enterotoxin (LT) were determined in only the environmental isolate 1J while non of the isolates produced heat stabile toxin (ST). The purification scheme was improved, few steps were adopted to include back extraction of ammonium sulfate, saturation between 80-20%, desalting through Sephadex G25, and gel filtration using Sephadex G100 which highly increase the specific activity.

scholarly journals Partial purification of thrombopoietin from the plasma of thrombocytopenic rabbits

Blood ◽  
1979 ◽  
Vol 54 (2) ◽  
pp. 377-388
Author(s):  
BL Evatt ◽  
J Levin ◽  
KM Algazy
Keyword(s):  
Ammonium Sulfate ◽  
Phosphate Buffer ◽  
Carboxymethyl Cellulose ◽  
Room Temperature ◽  
Deae Cellulose ◽  
Partial Purification ◽  
Elution Volume ◽  
Ammonium Sulfate Saturation ◽  
Citrate Phosphate ◽  
Citrate Phosphate Buffer

Partially purified thrombopoiesis-stimulating activity was prepared from the plasma of thrombocytopenic rabbits using ammonium sulfate precipitation and DEAE cellulose, Sephadex, and carboxymethyl cellulose chromatography. The protein fraction precipitated by an ammonium sulfate saturation of 60%-80%, previously shown to contain thrombopoiesis-stimulating activity, was used as starting material. Column chromatography was carried out at room temperature at pH 5.6. Under these conditions, thrombopoiesis-stimulating activity (thrombopoietin) was retained by DEAE cellulose (0/03 M citrate- phosphate buffer) and carboxymethyl cellulose (0/003 M citrate- phosphate buffer), and eluted with 0.4 M NaCl. Thrombopoietin was retarded by Sephadex G-100; the ratio of the elution volume to the void volume was 1.32:1. Immunoelectrophoretic analysis of partially purified thrombopoietin indicated that following removal of most of the albumin by DEAE chromatography, only proteins with the mobilities of beta- globulins and albumin and traces of other anodally migrating proteins were detectable in the fractions that contained thrombopoiesis- stimulating activity. Thrombopoietin was not dialyzable and was stable from at least pH 5.6 to 7.5. It was approximately 1000-fold purified following sequential chromatography with DEAE and carboxymethyl cellulose. Although the three fractions described reproducibly stimulated thrombopoiesis, as measured by increased levels of selenomethionine-75Se (75SeM) in the circulating platelets, platelet counts did not increase.

scholarly journals Ca2+-SPECIFIC REMOVAL OF Z LINES FROM RABBIT SKELETAL MUSCLE

1972 ◽  
Vol 52 (2) ◽  
pp. 367-381 ◽  
Author(s):  
Wayne A. Busch ◽  
M. H. Stromer ◽  
Darrel E. Goll ◽  
A. Suzuki
Keyword(s):  
Skeletal Muscle ◽  
Ammonium Sulfate ◽  
Protein Fraction ◽  
Saline Solution ◽  
Muscle Cells ◽  
The Other ◽  
Detectable Effect ◽  
Isoelectric Precipitation ◽  
Ammonium Sulfate Saturation

Removal of rabbit psoas strips immediately after death and incubation in a saline solution containing 1 mM Ca2+ and 5 nM Mg2+ for 9 hr at 37°C and pH 7.1 causes complete Z-line removal but has no ultrastructurally detectable effect on other parts of the myofibril. Z lines remain ultrastructurally intact if 1 mM 1,2-bis-(2-dicarboxymethylaminoethoxy)-ethane (EGTA) is substituted for 1 mM Ca2+ and the other conditions remain unchanged. Z lines are broadened and amorphous but are still present after incubation for 9 hr at 37°C if 1 mM ethylenediaminetetraacetate (EDTA) is substituted for 1 mM Ca2+ and 5 mM Mg2+ in the saline solution. A protein fraction that causes Z-line removal from myofibrils in the presence of Ca2+ at pH 7.0 can be isolated by extraction of ground muscle with 4 mM EDTA at pH 7.0–7.6 followed by isoelectric precipitation and fractionation between 0 and 40% ammonium sulfate saturation. Z-line removal by this protein fraction requires Ca2+ levels higher than 0.1 mM, but Z lines are removed without causing any other ultrastructurally detectable degradation of the myofibril. This is the first report of a protein endogenous to muscle that is able to catalyze degradation of the myofibril. The very low level of unbound Ca2+ in muscle cells in vivo may regulate activity of this protein fraction, or alternatively, this protein fraction may be localized in lysosomes.

Variability of the anticoagulant fraction of incubated fibrinogen

1965 ◽  
Vol 208 (3) ◽  
pp. 521-527 ◽  
Author(s):  
E. Triantaphyllopoulos ◽  
D. C. Triantaphyllopoulos
Keyword(s):  
Ammonium Sulfate ◽  
Continuous Flow ◽  
Early Stage ◽  
Paper Electrophoresis ◽  
Human Fibrinogen ◽  
Spontaneous Breakdown ◽  
Electrophoretic Patterns ◽  
Ammonium Sulfate Saturation ◽  
Advanced Stages

The spontaneous breakdown of human fibrinogen or the lysis of Laki's fibrinogen by purified human fibrinolysin and streptokinase involves at least two different reactions. Clottability by thrombin and precipitability at 25% ammonium sulfate saturation are gradually lost but although clottability disappears completely at a fairly early stage, precipitability is still retained by 4–12% of the original protein even during advanced stages. Both the amount and the antithrombic activity of the anticoagulant fraction of incubated fibrinogen (AFIF) produced vary throughout fibrinogenolysis. The changing quality of AFIF is further evidenced by changes in the electrophoretic patterns which indicate variations in concentration, mobility, and number of components. Strip electrophoresis of fractions isolated by continuous-flow paper electrophoresis showed the presence of two components in the largest fraction but only of a single one of different mobility in a second, smaller fraction. The last finding makes it unlikely that the antithrombic effect exerted by the latter fraction could be due to contamination by components of the former.

Physical, chemical, and enzymatic studies on the major sucrase of honey bees (Apis mellifera)

1976 ◽  
Vol 54 (2) ◽  
pp. 153-164 ◽  
Author(s):  
R. E. Huber ◽  
R. D. Mathison
Keyword(s):  
Apis Mellifera ◽  
Ammonium Sulfate ◽  
Honey Bees ◽  
Amino Sugar ◽  
Iodoacetic Acid ◽  
Subunit Structure ◽  
Ammonium Sulfate Saturation ◽  
Activity Loss ◽  
Substrate Concentrations ◽  
High Sucrose

A sucrase from honey bees (Apis mellifera) which precipitates between ammonium sulfate saturations of 50 and 70% (5 mg protein per millilitre) and which makes up the major portion of the sucrases of honey bees was purified to homogeneity as shown by several criteria. A large part of the sucrase was found in the head while most of the rest was in the abdomen (a small amount was in the thorax). The enzyme precipitated between the same values of ammonium sulfate saturation as did the sucrase in honey and honey sucrase exhibited kinetics very similar to those of this enzyme. The enzyme was found to be a relatively nonspecific α-glucosidase and was shown to have transglucosidase activity. The production of glucose from sucrose was rectilinear when plotted by the Hofstee method at low substrate concentrations but decreased at high sucrose concentrations. The production of fructose was rectilinear throughout the concentration range used. The production of both glucose and p-nitrophenol when p-nitrophenyl α-D-glucoside was the substrate was linear by the Hofstee plot. These effects were found to be due to transglucolysis and a mechanism of action is proposed. Amino acid and amino sugar analyses indicated that the sucrase was a glycoprotein. The molecular weight was found to be between 51 000 and 82 000 by three different methods and an s020,W value of 4.0 S was obtained. There was no evidence for subunit structure. Tests of the enzyme under various denaturation conditions did not reveal any unusual stabilities. The sucrase bound very tightly to a hydrophobic column. Iodoacetic acid decreased the activity of the sucrase but a large concentration was needed to bring about a 50% activity loss. Reducing agents caused some activity declines. Diethyl pyrocarbonate activated the enzyme.

scholarly journals Use of Aqueous Two-Phase and Three-Phase Partitioning Systems for Purification of Lipase Obtained in Solid-State Fermentation by Rhizopus arrhizus

2019 ◽  
Vol 13 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Valentina Dobreva ◽  
Boriana Zhekova ◽  
Georgi Dobrev
Keyword(s):  
Solid State ◽  
Ammonium Sulfate ◽  
Rhizopus Arrhizus ◽  
Two Phase ◽  
Sodium Tartrate ◽  
Phase Partitioning ◽  
Potassium Sodium ◽  
Three Phase ◽  
Ammonium Sulfate Saturation ◽  
Potassium Sodium Tartrate

Background: Purification of enzymes by conventional methods such as precipitation and chromatographic techniques is a costly and time-consuming procedure and may lead to low yields of enzyme activity. Alternative liquid-liquid extraction methods such as Aqueous Two-Phase Systems (ATPS) and Three Phase Partitioning (TPP) are characterized by the high enzyme yields and purification degree. Objective: The objective of this study was the application of partitioning systems ATPS and TPP for purification of lipase produced in solid-state fermentation by Rhizopus arrhizus. Methods: ATPS and TPP were used for purification of lipase, obtained by solid state cultivation of Rhizopus arrhizus. Results: Lipase was isolated with PEG4000/potassium sodium tartrate ATPS and the effect of the system composition, including PEG 4000 and potassium sodium tartrate concentrations on lipase partitioning was studied. When using 30% PEG4000/21% potassium sodium tartrate, lipase was distributed in the top phase, and the highest recovery yield of 217% and purification fold of 6.1 were achieved. It was found that at PEG4000 concentration of or higher than 15%, the enzyme was present in the top polymer-rich phase with a partitioning yield of over 90%. Upon application of TPP for lipase isolation, the effect of t-butanol concentration, ammonium sulfate concentration and pH on enzyme partitioning was investigated. The highest lipase recovery yield of 71% and 19.1-fold purification were achieved in the interfacial phase in the presence of 30% ammonium sulfate saturation with 1.0:0.5 crude extract/t-butanol ratio at pH 7 in a single step. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymographic analysis showed significant purification of lipase by TPP and the presence of two multiple forms of the enzyme. Conclusion: ATPS (PEG4000/ Potassium sodium tartrate) and TPP (1.0:0.5 crude extract/t-butanol ratio, 30% ammonium sulfate saturation, pH 7) proved to be rapid methods for the isolation and purification of lipase and they can be used in downstream processing for industrial preparation of the enzyme.

The Coagulation Abnormalities in Systemic Lupus Erythematosus

1968 ◽  
Vol 20 (03/04) ◽  
pp. 457-464 ◽  
Author(s):  
L Gonyea ◽  
R Herdman ◽  
R. A Bridges
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Ammonium Sulfate ◽  
Lupus Erythematosus ◽  
Anticoagulant Activity ◽  
Species Specificity ◽  
Sensitive Assay ◽  
Systemic Lupus ◽  
Coagulation Abnormalities

SummaryAn anticoagulant occurring in 4 of 6 patients with SLE has been demonstrated by a sensitive assay utilizing an ammonium sulfate fraction of serum. The anticoagulant functions as an inhibitor of the activation of prothrombin. No species specificity was demonstrable. The inhibitor behaves clinically and chromatographically as an immunoglobulin, although an attempt to demonstrate directly the antibody nature of the inhibitor was not successful.A severe, apparently independent, decrease in the level of prothrombin was observed in the patient with hemorrhagic symptoms. In contrast to the anticoagulant activity, the low prothrombin has persisted during treatment.

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