Fibrinolytic activities of euglobulins precipitated at pH 6.4, 6.0, and 5.3

1969 ◽  
Vol 47 (11) ◽  
pp. 935-940 ◽  
Author(s):  
A. M. Hedlin ◽  
F. C. Monkhouse
Keyword(s):  
Specific Activity ◽  
Deae Cellulose ◽  
Total Activity ◽  
Fibrinolytic System ◽  
Plasma Fibrinogen ◽  
Rabbit Plasma ◽  
Two Stage ◽  
The Difference ◽  
Ph Level ◽  
Cellulose Column

As a result of problems encountered with euglobulin preparation at pH 5.3, a study was undertaken to determine the pH level that provided the best recovery of fibrinolytic material. The euglobulin lysis test was used to measure the difference in activity at each pH level. Euglobulins were prepared from rabbit plasma at pH 6.4, 6.0, and 5.3, subjected to a variety of treatments, and separated on a DEAE cellulose column. Samples were assayed for plasminogen, plasmin, fibrinogen, prothrombin, and antithrombin levels. Results indicated the presence of plasminogen and plasmin at all three pH levels, with the greatest specific activity in the pH 6.0 and 5.3 euglobulins precipitated from defibrinated, urokinase-activated plasma. Maximum total activity was found in the pH 6.0 euglobulins. Approximately 25% of the plasma fibrinogen was precipitated at the pH 5.3 level. Trace amounts of prothrombin were found at all three pH levels. No measurable antithrombin was found in any of the samples. It was concluded that, of the three pH levels studied and with a two-stage euglobulin lysis test, the pH 6.0 precipitation provided the best measure of change in the fibrinolytic system.

Influence of dietary proteins on rennin and pepsin content of preruminant calf vell

1974 ◽  
Vol 41 (1) ◽  
pp. 19-23 ◽  
Author(s):  
Pascaline Garnot ◽  
E. Valles ◽  
J.-L. Thapon ◽  
R. Toullec ◽  
R. Tomassone ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Whey Proteins ◽  
Deae Cellulose ◽  
Milk Proteins ◽  
Total Activity ◽  
Separate Experiment ◽  
Dietary Proteins ◽  
Constant Weight ◽  
Cellulose Column ◽  
Qualitative Analyses

SummaryStudies were undertaken to determine the influence of dietary proteins on the rennin and pepsin contents of preruminant calf vell. Three groups of 12 Friesian calves were each fed either milk proteins, whey proteins or a 50:50 mixture of these 2 diets. They were slaughtered at a constant weight of 150kg and their vells collected and dried. Another group of vells was obtained from 8 animals that had been fed milk proteins in a separate experiment. The extraction of the abomasal enzymes was carried out at acid pH, and the extracts were quantitatively analysed for rennin and pepsin by DEAE-cellulose column chromatography. Qualitative analyses were also performed by agarose-acrylamide gel electrophoresis. The only enzymes observed using this last method were rennin and bovine pepsin II. Statistical analysis of the quantitative enzyme determinations indicated a trend for the vells from calves fed diets containing casein to be richer in total activity and in rennin, while the level of pepsin remained approximately constant. It seems that casein may induce the secretion of rennin. However, further experiments will be necessary to confirm this. Important differences were observed between the 2 groups of veils from calves given the same diet, but grown in slightly different conditions.

scholarly journals Isolation and characterization of a γ-type phosphoinositide-specific phospholipase C (PLC-γ2)

1990 ◽  
Vol 269 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Y Homma ◽  
Y Emori ◽  
F Shibasaki ◽  
K Suzuki ◽  
T Takenawa
Keyword(s):  
Phospholipase C ◽  
Column Chromatography ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Isolation And Characterization ◽  
Delta Type ◽  
Hydrolysis Of ◽  
Cellulose Column

A novel bovine spleen phosphoinositide-specific phospholipase C (PLC) has been identified with respect to immunoreactivity with four independent antibodies against each of the PLC isoenzymes, and purified to near homogeneity by sequential column chromatography. Spleen contains three of the isoenzymes: two different gamma-types [gamma 1 and gamma 2, originally named as PLC-gamma [Rhee, Suh, Ryu & Lee (1989) Science 244, 546-550] and PLC-IV [Emori, Homma, Sorimachi, Kawasaki, Nakanishi, Suzuki & Takenawa (1989) J. Biol. Chem. 264, 21885-21890] respectively] and delta-type of the enzyme, but PLC-gamma 1 is separated from the PLC-gamma 2 pool by the first DEAE-cellulose column chromatography. Subsequently, PLC-delta is dissociated on the third heparin-Sepharose column chromatography. The purified enzyme has a molecular mass of 145 kDa on SDS/polyacrylamide-gel electrophoresis and a specific activity of 12.8 mumol/min per mg with phosphatidylinositol 4,5-bisphosphate as substrate. This enzyme activity is dependent on Ca2+ for hydrolysis of all these phosphoinositides. None of the other phospholipids examined could be its substrate at any concentration of Ca2+. The optimal pH of the enzyme is slightly acidic (pH 5.0-6.5).

scholarly journals Purification and Characterization of Melanogenic Enzyme Tyrosinase from Button Mushroom

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Kamal Uddin Zaidi ◽  
Ayesha S. Ali ◽  
Sharique A. Ali
Keyword(s):  
Agaricus Bisporus ◽  
Specific Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Edible Mushroom ◽  
Total Activity ◽  
Protein Band ◽  
Exchange Chromatography ◽  
Ammonium Sulphate Precipitation ◽  
Key Enzyme

Melanogenesis is a biosynthetic pathway for the formation of the pigment melanin in human skin. A key enzyme, tyrosinase, catalyzes the first and only rate-limiting steps in melanogenesis. Since the discovery of its melanogenic properties, tyrosinase has been in prime focus and microbial sources of the enzyme are sought. Agaricus bisporus widely known as the common edible mushroom, it’s taking place in high amounts of proteins, enzyme, carbohydrates, fibers, and low fat contents are frequently cited in the literature in relation to their nutritional value. In the present study tyrosinase from Agaricus bisporus was purified by ammonium sulphate precipitation, dialysis followed by gel filtration chromatography on Sephadex G-100, and ion exchange chromatography on DEAE-Cellulose; the enzyme was purified, 16.36-fold to give 26.6% yield on total activity in the crude extract and final specific activity of 52.19 U/mg. The SDS-PAGE electrophoresis showed a migrating protein band molecular weight of 95 kDa. The purified tyrosinase was optimized and the results revealed that the optimum values are pH 7.0 and temperature 35°C. The highest activity was reported towards its natural substrate, L-DOPA, with an apparent Km value of 0.933 mM. This indicated that tyrosinase purified from Agaricus bisporus is a potential source for medical applications.

Studies on the relation between plasma antithrombin and heparin-cofactor

1968 ◽  
Vol 46 (3) ◽  
pp. 347-350 ◽  
Author(s):  
F. C. Monkhouse ◽  
Susan Milojevic
Keyword(s):  
Gel Electrophoresis ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Fold Increase ◽  
Significant Loss ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Patterns ◽  
Starch Gel ◽  
Cofactor Activity ◽  
Cellulose Column

A method for the preparation of purified plasma antithrombin and heparin-cofactor is described. The method involves adsorption by aluminium hydroxide, separation on a DEAE-cellulose column by means of a graded salt concentration, and vertical curtain electrophoresis. A 100-fold increase in the specific activity of antithrombin and a 30-fold increase in the specific activity of heparin-cofactor have been achieved. In spite of the increased purification, no separation of the two activities was achieved. When a highly purified fraction was subjected to starch-gel electrophoresis for 16–18 h and then eluted from the gel, there was significant loss of heparin-cofactor activity but not of antithrombin activity. The electrophoretic patterns of the recovered proteins were not altered.

scholarly journals Study on Isolation and Partial Purification of Lactase (β-galactosidase) Enzyme from Lactobacillus Bacteria Isolated from Yogurt

2011 ◽  
Vol 4 (1) ◽  
pp. 239 ◽  
Author(s):  
N. H. M. R. Mozumder ◽  
M. Akhtaruzzaman ◽  
M. A. Bakr ◽  
F. Tuj Zohra
Keyword(s):  
Specific Activity ◽  
Culture Media ◽  
Deae Cellulose ◽  
Chemical Properties ◽  
Total Activity ◽  
Partial Purification ◽  
Anion Exchange Column ◽  
Ammonium Sulphate Precipitation ◽  
Agar Media ◽  
Modified Media

Lactase has many applications in dairy industry including for the treatment of lactose intolerance. The present study was conducted to identify the activity of lactase enzyme produced by Lactobacillus bacteria isolated from yogurts available in Dhaka city. The strains were identified to be gram positive, catalase negative, fermentative and lactase producer when cultured on selective MRS agar media by using standard bacteriological procedures and techniques. The study revealed that enzymes produced by lactobacilli were capable to produce glucose from substrate lactose in lactose modified media using lactase assay Kit Glu IB and their highest protein concentration (17.25 mg/ml) was observed in the supernatant of culture media isolated from L. lactis. Highest total activity (850.69 U/l) and specific activity (50.04 U/mg) of lactase enzyme was observed in the strain of L. bulgaricus. The crude extract which showed highest activity was further purified by ammonium sulphate precipitation followed by anion exchange column chromatography (DEAE cellulose). Final specific activity and fold purification of lactase enzyme reached to 62.80 U/mg and 1.47 respectively. The highest physic-chemical properties (Effect of pH and temperature) of lactase enzyme were observed at PH 6.0 which was 43.98 U/mg of protein and at 70°c temperature which was 111.11 U/mg of protein.Keywords: β-Galactosidase; Specific activity; DEAE cellulose; Fold purification; Yogurt.© 2012 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved.doi: http://dx.doi.org/10.3329/jsr.v4i1.8478J. Sci. Res. 4 (1), 239-249 (2012)

scholarly journals The metabolism of cyclopentanol by Pseudomonas N.C.I.B. 9872

1972 ◽  
Vol 129 (3) ◽  
pp. 595-603 ◽  
Author(s):  
Martin Griffin ◽  
Peter W. Trudgill
Keyword(s):  
Specific Activity ◽  
Isocitrate Lyase ◽  
Deae Cellulose ◽  
Sole Source ◽  
Similar Rate ◽  
High Yield ◽  
Acetyl Coa ◽  
Ph Optimum ◽  
Nadph Oxidation ◽  
Cellulose Column

1. Pseudomonas N.C.I.B. 9872 grown on cyclopentanol as carbon source oxidized it at a rate of 228μl of O2/h per mg dry wt. and the overall consumption of 5.9μmol of O2/μmol of substrate. Cyclopentanone was oxidized at a similar rate with the overall consumption of 5.2μmol of O2μmol of substrate. Cells grown with sodium acetate as sole source of carbon were incapable of significant immediate oxidation of these two substrates. 2. Disrupted cells catalysed the oxidation of cyclopentanol to cyclopentanone by the action of an NAD+-linked dehydrogenase with an alkaline pH optimum. 3. A cyclopentanolinduced cyclopentanone oxygenase (specific activity 0.11μmol of NADPH oxidized/min per mg of protein) catalysed the consumption of 1μmol of NADPH and 0.9μmol of O2 in the presence of 1μmol of cyclopentanone. NADPH oxidation did not occur under anaerobic conditions. The only detectable reaction product with 100000g supernatant was 5-hydroxyvalerate. 4. Extracts of cyclopentanol-grown cells contained a lactone hydrolase (specific activity 7.0μmol hydrolysed/min per mg of protein) that converted 5-valerolactone into 5-hydroxyvalerate. 5. Cyclopentanone oxygenase fractions obtained from a DEAE-cellulose column were almost devoid of 5-valerolactone hydrolase and catalysed the formation of 5-valerolactone in high yield from cyclopentanone in the presence of NADPH. 6. Incubation of 5-hydroxyvalerate with the 100000g supernatant, NAD+ and NADP+ under aerobic conditions resulted in the consumption of O2 and the conversion of 5-hydroxyvalerate into glutarate. 7. The high activity of isocitrate lyase in cyclopentanol-grown cells suggests that the further oxidation of glutarate proceeds through as yet uncharacterized reactions to acetyl-CoA. 8. The reaction sequence for the oxidation of cyclopentanol by Pseudomonas N.C.I.B. 9872 is: cyclopentanol → cyclopentanone → 5-valerolactone → 5-hydroxyvalerate → glutarate → → acetyl-CoA.

Bovine Platelet Proteins III. Some Properties of Platelet Fibrinogen

1969 ◽  
Vol 21 (03) ◽  
pp. 428-440 ◽  
Author(s):  
N. O Solum ◽  
S Łopaciuk
Keyword(s):  
Intrinsic Viscosity ◽  
Rabbit Antiserum ◽  
Disc Electrophoresis ◽  
Carbohydrate Content ◽  
Plasma Fibrinogen ◽  
Total Amino Acid ◽  
Electrophoretic Mobilities ◽  
Bovine Plasma ◽  
Starch Gel ◽  
The Difference

Summary1. Some properties of purified bovine platelet fibrinogen have been described and the data compared to those obtained by parallel analysis of purified bovine plasma fibrinogen.2. A close similarity was found between platelet and plasma fibrinogen as to sedimentation coefficients, electrophoretic mobilities in starch gel and polyacrylamide disc electrophoresis, light absorption spectra in the range 240 mμ to 330 mμ, ability to form immunoprecipitate with a rabbit antiserum against bovine plasma fibrinogen, total amino acid composition and in N-terminal amino acids.Differences between the fibrinogens were found as to intrinsic viscosity, carbohydrate content and behaviour upon clotting by thrombin. Intrinsic viscosity in 0.3 M NaCl at 25° was 0.48 dl/g for platelet fibrinogen as compared to 0.26 dl/g for plasma fibrinogen. The carbohydrate content of platelet fibrinogen was 0.56 ± 0.10% 1.56±0.10% and 1.37±0.09% for sialic acid (calculated as N-glycolyl neuraminic acid), hexose (galactose/mannose 1:2) and hexosamine (glucosamine), respectively. These values were 6, 54 and 26% higher than those found for plasma fibrinogen. The difference in clotting behaviour indicated a slower polymerization rate of the fibrin monomers formed from platelet fibrinogen than of those formed from plasma fibrinogen.

Use of Factor VIII in the Treatment of Hemophilia

1962 ◽  
Vol 07 (02) ◽  
pp. 230-238 ◽  
Author(s):  
A Pavlovsky ◽  
H Peterson ◽  
G Casillas ◽  
C Simonetti ◽  
A Martinez Canaveri ◽  
...  
Keyword(s):  
Factor Viii ◽  
Tannic Acid ◽  
Deae Cellulose ◽  
Fibrinolytic System ◽  
Purification Method ◽  
Fraction I

SummaryThis publication describes the results obtained by treatment of haemophiliacs with factor VIII preparations isolated from Cohn fraction I by use of tannic acid, FI-O-Ta.The authors stress the rapidity of the disappearance of factor VIII after injection. Transfusions are generally well tolerated. One reaction of the pyrogen type has been observed and also a case of activation of the fibrinolytic system.A second purification method by means of chromatography on DEAE-cellulose is described.

Characterization of an isozyme of S-adenosylmethionine synthetase from rat liver

1984 ◽  
Vol 62 (5) ◽  
pp. 276-279 ◽  
Author(s):  
C. H. Lin ◽  
W. Chung ◽  
K. P. Strickland ◽  
A. J. Hudson
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Enzymatic Synthesis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Aromatic Amino Acids ◽  
Adenosylmethionine Synthetase ◽  
Cellulose Column

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.

Sign in / Sign up

Export Citation Format

Share Document