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.

CHANGES IN MILK, WHEY, AND BLUE CHEESE AS INDUCED BY BENZOYL PEROXIDE1,2

1974 ◽  
Vol 37 (5) ◽  
pp. 244-249 ◽  
Author(s):  
C. J. Washam ◽  
G. W. Reinbold ◽  
E. R. Vedamuthu ◽  
R. Jorgensen
Keyword(s):  
Heat Treatment ◽  
Benzoyl Peroxide ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Whey Proteins ◽  
Milk Proteins ◽  
Bleaching Process ◽  
Blue Cheese ◽  
Milk Whey ◽  
Heat Treated

Milk proteins were subjected to treatment with various levels of benzoyl peroxide, with and without heating at 60 C for 2 h. Heating had a pronounced effect on whey proteins, but polyacrylamide gel electrophoresis revealed changes in proteins not attributable to heat alone. The effect on proteins was reflected in an increased tendency for the benzoyl peroxide-heat treated cheeses to expel moisture during leakage tests. Use of 17.8 ppm benzoyl peroxide resulted in a markedly whiter cheese than that made using 5.9 ppm and reflectance studies indicated this to be true even when no heat treatment accompanied the benzoyl peroxide. Use of benzoyl peroxide in the bleaching process did not decrease mold development in ripening loaves nor was acid production by lactic cultures diminished. In addition, proteolysis of milk proteins by rennet was not reduced by the presence of benzoyl peroxide.

scholarly journals Chemical and immunochemical characterization of caseins and the major whey proteins of rabbit milk

1982 ◽  
Vol 201 (1) ◽  
pp. 71-79 ◽  
Author(s):  
R Dayal ◽  
J Hurlimann ◽  
Y M L Suard ◽  
J P Kraehenbuhl
Keyword(s):  
Amino Acid ◽  
Gel Electrophoresis ◽  
Phosphorus Content ◽  
Polyacrylamide Gel Electrophoresis ◽  
Whey Proteins ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Precipitation Line ◽  
Specific Antibodies ◽  
Rabbit Milk

Caseins were separated from whey proteins by acid precipitation of skimmed rabbit milk. Whole casein was resolved by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis into three major bands with apparent relative molecular masses (Mr of 31 000, 29 000 and 25 000. On agarose/urea-gel electrophoresis whole casein gave three bands with electrophoretic mobilities alpha, beta and gamma. The three components were purified by DEAE-cellulose chromatography under denaturing and reducing conditions. Each was shown to have a different amino acid, hexose and phosphorus content, as well as non-identical peptide fragments after proteinase digestion. The 31 000 Da (dalton) protein, of alpha-electrophoretic mobility, had a high phosphorus content (4.38%, w/w); the 29 000 Da peptide, of gamma-mobility, had the highest hexose content (2.2%, w/w), contained 0.8 cysteine residue per 100 amino acid residues and was susceptible to chymosin digestion corresponding thus to kappa-casein; the 25 000 Da protein migrated to the beta-position. The rabbit casein complex is composed of at least three caseins, two of which (alpha- and kappa-caseins) are analogous to the caseins from ruminants. Although caseins are poor immunogens, specific antibodies were raised against total and purified polypeptides. The antiserum directed against whole casein recognized each polypeptide, each casein corresponding to a distinct precipitation line. The antisera directed against each casein polypeptide reacted exclusively with the corresponding casein and no antiserum cross-reaction occurred between the three polypeptides. From whey, several proteins were isolated, characterized and used as antigens to raise specific antibodies. An iron-binding protein with an apparent Mr of 80 000 was shown to be immunologically and structurally identical with serum transferrin.

HUMAN SERUM PRECIPITINS TO GOAT'S MILK PROTEINS AND TO COW'S MILK PROTEINS

PEDIATRICS ◽  
1965 ◽  
Vol 35 (2) ◽  
pp. 247-253
Author(s):  
Melvin Lee
Keyword(s):  
Serum Proteins ◽  
Whey Proteins ◽  
Deae Cellulose ◽  
Milk Proteins ◽  
Double Diffusion ◽  
Cow’S Milk ◽  
Cow's Milk ◽  
Goat’S Milk ◽  
Human Sera ◽  
Goat's Milk

Antigenic relationships of cow's milk, cow's serum, goat's milk, and goat's serum were studied, using techniques of agar double-diffusion, DEAE-cellulose fractionation of milk proteins, and human sera containing precipitins to cow's milk. Immunologically similar proteins are present in goat's serum and cow's serum, and evidence is presented for the occurrence of crossreacting antigenic serum proteins in milk of both species. However, the samples of milk also contain other proteins, unreleated to serum proteins, which react with rabbit and human antisera. Several human sera which contained antibodies to cow's milk were studied. Antibodies to goat's milk could be demonstrated in most of these, and to cow's serum and goat's serum in some of them. No precipitin lines were formed when rabbit or human sera were tested against commercial evaporated goat's milk, although precipitin lines were formed with reconstituted commercial dried goat's milk. Milk samples were separated into 16 fractions by column chromatographic procedures and tested against human sera. One serum sample contained antibodies only to whey proteins while two other sera contained antibodies to all 16 fractions in both cow's milk and goat's milk.

scholarly journals Purification and characterization of biotin-binding protein II from chicken oocytes

1988 ◽  
Vol 256 (3) ◽  
pp. 797-805 ◽  
Author(s):  
L Bush ◽  
T J McGahan ◽  
H B White
Keyword(s):  
Amino Acid ◽  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Chicken Egg ◽  
Biotin Binding ◽  
Cellulose Column

BBP-II, the major biotin-binding protein from chicken oocytes, was purified 12,000-fold with a 22% yield. The purification procedure includes butan-1-ol extraction of yolk lipids, phosphocellulose chromatography of the water-soluble proteins, DEAE-cellulose chromatography at pH 7.4 and hydroxyapatite column chromatography. Final purification was obtained by using a second DEAE-cellulose column chromatography at pH 6.0. BBP-I activity separated from BBP-II activity during elution from the first DEAE-cellulose column. Purified BBP-II was homogeneous on both polyacrylamide-gel electrophoresis and SDS/polyacrylamide-gel electrophoresis under conditions that would detect a 1% impurity. The subunit Mr determined from SDS/polyacrylamide-gel electrophoresis was 18,200 (72,600 for tetramer), which compares favourably with an Mr value of 17,300 (69,100) calculated from the amino acid analysis. A single precipitin line formed when rabbit antiserum to the protein was directed against a crude chicken egg-yolk sample. BBP-II purified by this procedure lacked carbohydrate and phosphate, was stable indefinitely when frozen, and was quite stable at room temperature. The N-terminal amino acid sequence showed polymorphism at three positions in the first 23 residues and was about 45% identical with the N-terminal 22 residues of avidin. Antiserum to BBP-II cross-reacted with BBP-I and similar proteins in the yolk of eggs from various birds and alligator as judged by immunodiffusion and enzyme-linked immunosorbent assays. No cross-reaction was observed with chicken egg-white by either of these methods.

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.

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.

scholarly journals Purification and chemical characterization of human hexosaminidases A and B

1976 ◽  
Vol 159 (3) ◽  
pp. 535-539 ◽  
Author(s):  
J E S. Lee ◽  
A Yoshida
Keyword(s):  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Gel Chromatography ◽  
Deae Cellulose Column ◽  
Hexosaminidase A ◽  
Cellulose Column

N-Acetyl-β-hexosaminidases A and B were purified to homogeneity from human placenta. In the initial step of purification, the enzymes were adsorbed on concanavalin A-Sepharose 4B and eluted from the column with α-methyl D-mannosides. Subsequent purification steps included DEAE-cellulose column chromatography, QAE-Sephadex [diethyl-(2-hydroxypropyl)aminoethyl-Sephadex] column chromatography, Sephadex G-200 gel filtration and preparative disc polyacrylamide-gel electrophoresis, followed by another QAE-Sephadex chromatography for the hexosaminidase A preparation, and DEAE-cellulose column chromatography, calcium phosphate gel chromatography, Sephadex G-200 gel filtration, QAE-Sephadex chromatography and CM-cellulose chromatography for the hexosaminidase B preparation. The purified preparations, particularly hexosaminidase A, had significantly higher specific enzyme activities than previously reported. The preparations moved on polyacrylamide-gel electrophoresis as single protein bands, which also stained for enzyme activity. Sedimentation-equilibrium centrifugation indicated homogenous dispersion of the enzymes, and the molecular weight was estimated as about 110000 for both enzymes. Complete amino acid and carbohydrate compositions of the two isoenzymes were determined, and, in contrast with previous suggestions, no sialic acid was found in the enzymes.

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