scholarly journals The purification and properties of butyryl-coenzyme A dehydrogenase from Peptostreptococcus elsdenii

1971 ◽  
Vol 125 (3) ◽  
pp. 879-887 ◽  
Author(s):  
Paul C. Engel ◽  
V. Massey
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Simple Procedure ◽  
Potassium Bromide ◽  
Prosthetic Group ◽  
Pig Liver ◽  
Peak Absorption ◽  
Purification And Properties ◽  
Green Form ◽  
Spectrophotometric Titrations

Butyryl-CoA dehydrogenase prepared by a simple procedure from Peptostreptococcus elsdenii has a molecular weight of approx. 150000. The enzyme has FAD as its prosthetic group. The amino acid analysis is reported. This enzyme, like most of the corresponding mammalian ones, is green. The absorption band at 710nm can be abolished irreversibly by dithionite reduction and air reoxidation; it can be abolished reversibly by phenylmercuric acetate or potassium bromide. The enzyme as isolated appears to be a mixture of a green and a yellow form, both of which are active. This view is supported by the variable ‘greenness’ of different preparations and the biphasic curve obtained in anaerobic spectrophotometric titrations with dithionite. It can be calculated from the titration results that fully green enzyme would have a peak-to-peak absorption ratio (E710/E430) as great as 0.54. The green form is much less rapidly reduced by dithionite than the yellow form, but is nevertheless much more readily reduced by dithionite than the enzyme from pig liver. It is also more readily reoxidized by air and shows less tendency to form a semiquinone. Treatment with sodium borohydride produces an unusual reduced species that is probably the 3,4-dihydroflavin.

scholarly journals Purification and properties of the nicotinamide–adenine dinucleotide phosphate-dependent isocitrate dehydrogenase from pig liver cytoplasm

1970 ◽  
Vol 118 (2) ◽  
pp. 253-258 ◽  
Author(s):  
J. A. Illingworth ◽  
K. F. Tipton
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Isocitrate Dehydrogenase ◽  
Nicotinamide Adenine Dinucleotide ◽  
Soluble Fraction ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Turnover Number ◽  
Pig Liver ◽  
Extinction Coefficients ◽  
Purification And Properties

The NADP-dependent isocitrate dehydrogenase from pig liver soluble fraction was purified over 500-fold with an overall yield of 25%. The purified enzyme, which is homogeneous by all the usual criteria, has a molecular weight of about 75000 and is composed of two identical subunits. This has been demonstrated by ultracentrifugation, fluorescence titration and peptide `fingerprinting'. The maximal turnover number, extinction coefficients at 280nm and 260nm and amino acid analysis are described.

scholarly journals Purification and Properties of Staphylocoagulase

1975 ◽  
Author(s):  
A.D. Muller ◽  
B. M. Bas ◽  
H. C. Hemker
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Aspartic Acid ◽  
Acid Composition ◽  
Isoelectric Point ◽  
Terminal Amino Acid ◽  
Purification And Properties ◽  
Terminal Amino

Staphylocoagulase, an exoprotein of coagulase positive staphylocoagulase, has been purified to a state in which only trace amounts of contaminating proteins are detectable.Purification was more than 35,000 fold, which is 7 times more than the highest value reported in the literature. The yield was about 15%.Aspartic acid was found as a single N-terminal amino acid in this preparation. The molecular weight is 61,000 and the isoelectric point lies at pH 4.53.The amino acid composition was determined.

Purification and properties of crustacyanin

1966 ◽  
Vol 164 (994) ◽  
pp. 130-151 ◽  
Keyword(s):  
Molecular Weight ◽  
Quaternary Structure ◽  
Gel Filtration ◽  
Potassium Thiocyanate ◽  
Starch Gel Electrophoresis ◽  
Prosthetic Group ◽  
Reversible Dissociation ◽  
Purification And Properties ◽  
Starch Gel ◽  
Quaternary Structures

Crustacyanin, the blue carapace pigment of the common lobster Homarus gammarus (L.), has been purified and crystallized. This chromoprotein has a minimum molecular weight of 36 000 based on the content of the carotenoid prosthetic group astaxanthin. The molecular weight in gel filtration measurements is about 650 000, corresponding to some 18 molecules of astaxanthin per molecule of protein. Crustacyanin, on dialysis against water, dissociates into particles of about 35 000 molecular weight, each apparently bearing one molecule of carotenoid. The dissociation is accompanied by a shift in the principal maximum of the absorption spectrum from 633 to 595 nm and is reversed upon addition of salt. Reversible dissociation also occurs in the presence of 3 M urea, 1 M potassium thiocyanate, 10% (v/v) dioxan or 10% (v/v) acetone. When the carotenoid is removed from crustacyanin with acetone, the resul­tant apoprotein has a mean molecular weight of about 20 000. It may be resolved by starch gel electrophoresis into several components of which two predominate. Crustacyanin, indistinguishable from the native material, can be reconstituted from apoprotein and carotenoid. Evidence from the behaviour of crustacyanin and its apoprotein at surfaces indicates that the tertiary and quaternary structures of the native protein are stabilized by the carotenoid. It is suggested that the quaternary structure of crustacyanin is induced by an interaction of the carotenoid molecules of the subunits, which in turn causes a change in configuration of the protein favourable to aggregation. The result is a micelle-like structure with a hydrophobic carotenoid core.

Purification and properties of a phenol carboxylic acid acyl esterase from Aspergillus flavus

1971 ◽  
Vol 17 (11) ◽  
pp. 1455-1463 ◽  
Author(s):  
J. J. Child ◽  
T. Oka ◽  
F. J. Simpson ◽  
H. G. Krishnamurty
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Carboxylic Acid ◽  
Aspergillus Flavus ◽  
Protocatechuic Acid ◽  
Hydroxybenzoic Acid ◽  
Purification And Properties ◽  
Wide Range ◽  
Hydrolysis Of ◽  
Active Preparation

Aspergillus flavus produces an extracellular esterase that hydrolyses phenolic carboxylic acid acyl esters. An assay based upon the measurement of the rate of release of phloroglucinol on hydrolysis of the ester of phloroglucinol and protocatechuic acid is described. The most active preparation hydrolyzed 30.8 μmoles of substrate per minute per milligram of protein and was active against a wide range of esters of meta and para hydroxybenzoic acid derivatives.The enzyme was isolated as a homogeneous protein, as judged by ultracentrifugation and by electrophoresis and had an isoelectric point of pH 4.45. The molecular weight was determined as 166 000. The enzyme is a glycoprotein containing 42.8% carbohydrate and the amino acid composition is described.

scholarly journals Colouring Matters of the Aphidoidea XLII. Purification and Properties of the Cyclising Enzyme [Protoaphin Dehydratase (Cyclising)] Concerned with Pigment Transformations in the Woolly Aphid Eriosoma lanigerum Hausmann (Hemiptera: Insecta)

1977 ◽  
Vol 30 (3) ◽  
pp. 173 ◽  
Author(s):  
DW Cameron ◽  
WH Sawyer ◽  
VM Trikojus
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Analysis ◽  
Eriosoma Lanigerum ◽  
Purification And Properties ◽  
Woolly Aphid

Dried extracts of woolly aphid were treated with n-butanol, then by chromatography on DEAESephadex A50 and finally by filtration on Sephadex G150 to yield a substantially homogeneous protein catalysing the conversion of the aglucone of protoaphin into xanthoaphin. Traces of lowmolecular- weight contaminants were removed by chromatography on Sephadex G 100. The enzyme, which has a molecular weight of 120000�2000 and a high content of p-structure, was inhibited by naphthoresorcinol. Its glycoprotein nature was indicated by amino acid analysis.

scholarly journals The purification and properties of the l-serine O-sulphate-degrading system of pig liver

1971 ◽  
Vol 121 (5) ◽  
pp. 747-752 ◽  
Author(s):  
N. Tudball ◽  
P. Thomas ◽  
R. Bailey-Wood
Keyword(s):  
Amino Acid ◽  
Isoelectric Focusing ◽  
Enzyme System ◽  
Gel Filtration ◽  
Pig Liver ◽  
Molecular Weights ◽  
Purification And Properties ◽  
Isoelectric Points ◽  
Degrading System ◽  
Similar Amino Acid

1. The enzyme system from pig liver responsible for the αβ-elimination of l-serine O-sulphate was purified 1000-fold. 2. Isoelectric focusing produced two enzymically active fractions with isoelectric points at pH5.6 and 5.9 respectively. 3. Osmometry and gel filtration showed both enzymes to possess molecular weights of approx. 54000. 4. The separate activities exhibited similar amino acid compositions.

Purification and Properties of γ-Glutamyl Cyclotransferase from Pig Liver

1971 ◽  
Vol 49 (2) ◽  
pp. 218-226 ◽  
Author(s):  
E. D. Adamson ◽  
A. Szewczuk ◽  
G. E. Connell
Keyword(s):  
Amino Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ammonium Sulfate Precipitation ◽  
Cyclic Derivative ◽  
Pig Liver ◽  
Molecular Weights ◽  
Purification And Properties ◽  
Isoelectric Points ◽  
Pyrrolidone Carboxylic Acid ◽  
Amino Acid Glycine

The widely occurring enzyme γ-glutamyl cyclotransferase acts on γ-glutamyl peptides to effect the release of the terminal glutamyl residue as the cyclic derivative pyrrolidone carboxylic acid. The enzyme has been purified from pig liver by (1) ammonium sulfate precipitation from the supernatants of homogenates, (2) CM-cellulose treatment, (3) DEAE-Sephadex chromatography, and (4) preparative polyacrylamide gel electrophoresis. The homogeneity of the highly purified enzyme was demonstrated by ultracentrifugation and electrophoretic analyses. By means of (5) isoelectric focusing, two forms of the enzyme with isoelectric points 4.87 and 4.95 were separated. These forms proved to have very similar sedimentation velocities, molecular weights, and amino acid compositions, and to have the same amino acid, glycine, as the N-terminal residue.

Acetylcholinesterase aus dem Gift von Bungarus multicinctus. Reinigung und Eigenschaften/The Acetylcholinesterase of Bungarus multicinctus Venom. Purification and Properties

1979 ◽  
Vol 34 (1-2) ◽  
pp. 27-32 ◽  
Author(s):  
H. Großmann ◽  
M. Weinert ◽  
M. Liefländer
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Amino Acid Analysis ◽  
Specific Activity ◽  
Terminal Amino Acid ◽  
Purification And Properties ◽  
Gradient Gel Electrophoresis ◽  
Terminal Amino

Abstract Acetylcholinesterase from Banded krait (Bungarus multicinctus) venom has been purified by CM-Sephadex chromatography and affinity chromatography to a specific activity of 4290 U/mg. The purified enzyme is a glycoprotein. It is free of electrophoretically detectable contaminating proteins. A molecular weight of 140 000 ± 5 000 has been determined by gradient gel electrophoresis for the native enzyme. It is split into two equal-sized subunits (Mr 70 000 ± 2 000) by SDS treatment. The N-terminal amino acid analysis gave glycine and serine. The purified acetyl­ cholinesterase can be resolved by disc gel electrophoresis into four and by isoelectric focusing into six isozymes. The pI value of the main isozyme has been found to be 5.98 ± 0.05.

scholarly journals Molecular properties of lupin and serradella leghaemoglobins

1972 ◽  
Vol 127 (1) ◽  
pp. 309-314 ◽  
Author(s):  
W. J. Broughton ◽  
M. J. Dilworth ◽  
C. A. Godfrey
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Cellulose Acetate ◽  
Root Nodules ◽  
Deae Cellulose ◽  
Lupinus Luteus ◽  
Terminal Amino Acid ◽  
Prosthetic Group ◽  
Terminal Amino ◽  
Ornithopus Sativus

1. Leghaemoglobins were extracted from the root nodules of lupin (Lupinus luteus L.) and serradella (Ornithopus sativus Brot.) plants and fractionated into different leghaemoglobin components on DEAE-cellulose–acetate columns. 2. The first two fractions eluted from columns loaded with either lupin or serradella leghaemoglobins were in the Fe3+ oxidation state. 3. These components have protohaem IX as the prosthetic group and glycine as the N-terminal amino acid. 4. Other properties are: lupin component I, pI5.08, molecular weight 19000; lupin component II, pI5.13, molecular weight 20600; serradella component I, pI5.00, molecular weight 17500; serradella component II, pI5.05, molecular weight 19100. 5. Leghaemoglobins are thus heterogeneous with respect to size and charge.

The function of fimbriae in Myxococcus xanthus. I. Purification and properties of M. xanthus fimbriae

1979 ◽  
Vol 25 (10) ◽  
pp. 1152-1160 ◽  
Author(s):  
W. J. Dobson ◽  
H. D. McCurdy
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Amino Acid ◽  
Myxococcus Xanthus ◽  
Hydrolytic Enzymes ◽  
Antigenic Determinants ◽  
Freeze Thaw ◽  
Purification And Properties ◽  
Precise Relationship ◽  
Type Studies

Myxococcus xanthus fimbriae have been purified and characterized as part of a study of the function of fimbriae in this prokaryote. Myxococcus xanthus produced two types of fimbriae, termed flaccid (F) and rigid (R) on the basis of electron microscopy. F and R fimbriae differed slightly in their response to pH and freeze–thaw regimes but were similar in their resistance to hydrolytic enzymes, amino acid composition, molecular weight, carbohydrate content, and antigenic determinants. Although the precise relationship between F and R fimbriae is unknown, the possibility is considered that F fimbriae might represent a "contracted" form of the R type. Studies designed to determine fimbriae function in M. xanthus are described in an accompanying report.

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