scholarly journals The cellulase of Fusarium solani. Purification and specificity of the β-(1→4)-glucanase and the β-d-glucosidase components

1971 ◽  
Vol 121 (3) ◽  
pp. 353-362 ◽  
Author(s):  
T. M. Wood
Keyword(s):  
Heat Treatment ◽  
Molecular Weight ◽  
Phosphoric Acid ◽  
Isoelectric Focusing ◽  
Fusarium Solani ◽  
Reducing Sugars ◽  
Activity Ratio ◽  
Gel Filtration ◽  
Cellulase Activity ◽  
Isoelectric Points

1. Cell-free culture filtrates of the fungus Fusarium solani were examined for homogeneity with respect to β-d-glucosidase and Cx activities. 2. o-Nitrophenyl β-d-glucoside and cellobiose were both used as substrates for β-d-glucosidase activity. 3. No evidence for the non-identity of nitrophenyl β-d-glucosidase and cellobiase activities could be found, either by heat treatment, gel filtration on Sephadex G-100 or by isoelectric focusing. 4. The β-d-glucosidase component was also a feeble exo-β-glucanase: it had a molecular weight of approx. 400000. 5. The fall in viscosity of a solution of CM-cellulose, the formation of reducing sugars in a solution of CM-cellulose and the solubilization of phosphoric acid-swollen cellulose (Walseth cellulose), were all used for the measurement of Cx activity. 6. The ratio of the two types of CM-cellulase activity was not changed after gel filtration on Sephadex G-100 or after chromatography on DEAE-Sephadex. 7. Three peaks of Cx activity were obtained after electrofocusing, but all three possessed the same ratio of the two types of CM-cellulase activity as well as the same CM-cellulase/Walseth activity ratio, as the unfractionated enzyme; all three isoenzymes (isoelectric points, 4.75, 4.80–4.85 and 5.15) acted in synergism with a mixture of the C1 and the β-d-glucosidase components to the same extent in the solubilization of cotton fibre. 8. The molecular weight of the Cx component was approx. 37000.

A cellulase complex in culture filtrates of Penicillium citrinum

1976 ◽  
Vol 22 (8) ◽  
pp. 1153-1159 ◽  
Author(s):  
P. O. Olutiola
Keyword(s):  
Molecular Weight ◽  
Reducing Sugars ◽  
Gel Filtration ◽  
Cellulase Activity ◽  
Cellulase Production ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Penicillium Citrinum ◽  
Low Molecular Weight ◽  
Weight One

During growth in a liquid medium that contained a single soluble or an insoluble cellulosic carbon source Penicillium citrinum released a complex of cellulase enzymes into the medium. A temperature of 30 °C was best for cellulase production. Presence of carbon-containing compounds, particularly glucose, inhibited cellulase activity. The enzyme complex was separated by gel filtration followed by ion-exchange chromatography into 11 components, 4 of high molecular weight and 7 of low molecular weight. One of the components (Bb) had the character of C1 cellulase enzyme. When the components were combined they released more reducing sugars from cullulosic substrates than when they were used singly.

scholarly journals Rat small-intestinal β-galactosidases. Studies on the fractionation of ‘acid’ β-galactosidase with isoelectric focusing, gel filtration and ion-exchange chromatography

1970 ◽  
Vol 117 (2) ◽  
pp. 369-375 ◽  
Author(s):  
Nils-Georg Asp
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
High Molecular Weight ◽  
Isoelectric Focusing ◽  
Gel Filtration ◽  
Ion Exchange Resin ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Isoelectric Points ◽  
Small Intestinal

1. Different forms of the rat small-intestinal ‘acid’ β-galactosidase were separated by using the isoelectric-focusing technique. The isoelectric points of the different forms were at pH4.2, 4.6, 5.4, 6.1 and approx. 8. 2. The two forms of ‘acid’ β-galactosidase isoelectric at pH4.2 and 4.6 were completely excluded from the Sephadex G-200 gel, whereas the form isoelectric at pH8 had Kav. 0.4. The concentration and pH of the elution buffer influenced the distribution of enzyme activity between different forms. Thus, under certain conditions of ionic strength and pH, the enzyme seems to form high-molecular-weight aggregates with low isoelectric points. These may be homopolymeric aggregates or the result of binding of enzyme to, for example, membrane fragments. The forms isoelectric at pH5.4 and 6.1 are probably aggregates of intermediate size. 3. During ion-exchange chromatography at pH6.0 one fraction of ‘acid’ β-galactosidase was not retained on the column and was isoelectric at pH8 and another fraction was eluted when the buffer concentration in the eluate had increased to about 50mm. The main part of enzyme eluted in this second fraction was also isoelectric at pH8, indicating that the elution of this fraction is not a simple ion-exchange procedure but probably also involves a splitting of high-molecular-weight aggregates, originally retained because of their low isoelectric points. The enzyme subunits have a higher isoelectric point, and are therefore no longer bound to the ion-exchange resin.

Purification of a Glycoprotein from Bovine Leukemia Virus (BLV)

1977 ◽  
Vol 32 (3-4) ◽  
pp. 301-304 ◽  
Author(s):  
B. Frenzel ◽  
. R. Kaaden ◽  
M. Mussgay
Keyword(s):  
Molecular Weight ◽  
Isoelectric Focusing ◽  
Bovine Leukemia Virus ◽  
Gel Electrophoresis ◽  
Leukemia Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Relative Molecular Weight ◽  
Bovine Leukosis

Abstract A precipitating antigen of bovine leukemia virus was isolated by isoelectric focusing and Sephadex gel filtration. In SDS-polyacrylamide gel electrophoresis it was found to be a homogeneous protein with a relative molecular weight of 69 000 daltons. Because of its relative molecular weight and staining characteristics it was designated as BLV gp69. A protein with the same molecular weight could also be demonstrated in BLV particles. In 34 out of 35 sera from cattle affected by enzootic bovine leukosis antibodies against gp69 were detected, whereas the sera from 197 animals, free of bovine leukosis, did not react in immunodiffusion test.

Renin Precursor and Its Activation Mechanism in Hog Kidney

1980 ◽  
Vol 59 (s6) ◽  
pp. 21s-24s ◽  
Author(s):  
Kazuo Murakami ◽  
Saori Takahashi ◽  
Shigehisa Hirose ◽  
Yukio Takii ◽  
Tadashi Inagami
Keyword(s):  
Molecular Weight ◽  
Affinity Chromatography ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Activation Mechanism ◽  
Net Charge ◽  
Isoelectric Points ◽  
Binding Substance ◽  
Inactive Renin ◽  
Hog Kidney

1. A completely inactive renin was isolated from hog kidney extract by affinity chromatography on pepstatin-aminohexyl-Sepharose and on an Affi-Gel Blue column. 2. This inactive renin had a molecular weight of 43 000 ± 1500 as determined by gel filtration on Ultrogel AcA 44. Upon activation with trypsin, its molecular weight fell to 41 000 ± 1400. 3. The inactive renin lacked the ability to bind renin-binding substance whereas trypsin-activated renin was able to bind the renin-binding protein and to form high-molecular-weight renin. 4. Chymotrypsin as well as trypsin could activate the inactive renin although less effectively. 5. The active renins generated from the inactive renin by the action of different proteolytic enzymes differed in their net charge, reflecting the specificities of the proteases used; the isoelectric points of the native, the trypsin-activated and the chymotrypsin-activated forms of renin occurred at pH 5.3, 5.1 and 4.8 respectively.

A comparison of four sulfhydryl cathepsins (B, C, H, and L) from porcine spleen

1984 ◽  
Vol 62 (12) ◽  
pp. 1301-1308 ◽  
Author(s):  
K. R. Lynn ◽  
Rosalind S. Labow
Keyword(s):  
Molecular Weight ◽  
High Pressure ◽  
Cathepsin L ◽  
Gel Filtration ◽  
Cathepsin C ◽  
Molecular Weights ◽  
Isoelectric Points ◽  
Dipeptidyl Aminopeptidase ◽  
Precise Assessment ◽  
Similar Amino Acid

Four sulfhydryl cathepsins, B, C (dipeptidyl aminopeptidase I), H, and L were isolated from porcine spleen. They are all glycoproteins of similar amino acid compositions, which are comparable with those of cathepsins B and H from other sources and so with papain. All four cathepsins exist in multiple charged forms: B, C, H, and L have isoelectric points in the range 4.3–5.4, 5.3 and 5.9, 5.2–5.7, and 7–8.7, respectively. The molecular weights of cathepsins B and H were 24 000 and 26 000. Anomalous behaviour of cathepsin L on both conventional gel filtration and high pressure liquid chromatography precluded a precise assessment of its weight which is between 22 000 and 28 000. The isolated mercurial derivative of cathepsin C has a molecular weight of 56 000 (an active dimer formed on reduction). Cathepsins B and H also aggregate.

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.

scholarly journals Polypeptide Composition of Arachin and Non-arachin Proteins From Early Bunch Peanut (Arachis hypogaea L.) Seed1

1981 ◽  
Vol 8 (2) ◽  
pp. 82-88 ◽  
Author(s):  
Shaik-M. M. Basha ◽  
Sunil K. Pancholy
Keyword(s):  
Molecular Weight ◽  
Arachis Hypogaea ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Seed Proteins ◽  
Two Dimensional ◽  
Polypeptide Composition ◽  
Two Dimensional Gel Electrophoresis ◽  
Arachis Hypogaea L ◽  
Isoelectric Points

Abstract Peanut (Arachis hypogaea L.) seed proteins were resolved into arachin and non-arachin fractions, and composite two-dimensional polypeptide maps were prepared. Seed proteins were extracted with a buffer containing 2 M NaCl, 10 mM Tris-HCl (pH 8.2), 0.2 mM phenylmethyl sulfonyl fluoride and 0.002% NaN3 and resolved into ten peaks by gel filtration on a Sephacryl S-300 column. Gel filtration of total protein extract yielded three molecular weight variants (490,000., 400,000, and 365,000) of arachin. Gel electrophoresis showed quantitative and qualitative differences in the protein and polypeptide composition of the three arachin variants. Nonarachin proteins obtained by this method were heterogeneous and distinct from the arachin. Two-dimensional gel electrophoresis revealed several differences in the polypeptide composition between arachin fraction IV and fractions II and III. Composite two-dimensional polypeptide maps of arachin and non-arachin revealed the presence of several polypeptides with similar isoelectric points and molecular weights between them. Arachin contained six molecular weight (between 15,500 and 68,000) classes of polypeptides with isoelectric points between 4.7 and 8.4 while nonarachin contained nine molecular weight (between 16,000 and 170,000) classes of polypeptides having isoelectric points between 4.7 and 7.9.

Separation and Characterization of Two Fibrinolytic Inhibitors from Human Placenta

1971 ◽  
Vol 25 (03) ◽  
pp. 580-589 ◽  
Author(s):  
M Uszynski ◽  
U Abildgaard
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Isoelectric Focusing ◽  
Basic Protein ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Coagulation Inhibitor ◽  
Tissue Thromboplastin

SummaryProcedures for the separation of two inhibitors of the activation of plasminogen to plasmin by urokinase are described. Tissue thromboplastin was removed by adsorption to Al(0H)3 gel followed by ultracentrifugation. Plasminogen, plasminogen activator, a coagulation inhibitor and hemoglobin were removed by ion exchange chromatography (CM- or DEAE-Sephadex with NaCl gradients). The minor UK inhibitor is a relative basic protein with a pI of about 5.8. The major inhibitor was purified further by isoelectric focusing, preparative electrophoresis in polyacrylamide gel, and gel filtration. This inhibitor has α1-motility, the pI is about 5.2, and the molecular weight about 100,000. It inactivates urokinase progressively, but does not inhibit streptokinase, plasmin or thrombin.

scholarly journals Characterization of the estrogen-induced uterine peroxidase by microelectrophoresis.

1978 ◽  
Vol 26 (5) ◽  
pp. 382-390 ◽  
Author(s):  
C C Phillips Burnett ◽  
W A Anderson ◽  
R Rüchel
Keyword(s):  
Molecular Weight ◽  
Isoelectric Focusing ◽  
Large Subunit ◽  
The Other ◽  
Two Dimensional ◽  
Neuraminidase Treatment ◽  
Gradient Electrophoresis ◽  
Isoelectric Points ◽  
Uterine Fluid

Estrogen-dependent peroxidase from rat uterine fluid has been investigated by microelectrophoretic techniques. The molecular weight of the enzyme has been determined in the range of 100,000 by using polyacrylamide gradient gels in the absence and presence of nonionic and anionic detergent. The isoelectric points are located between pH 4.5 and 5.9. Employing the two-dimensional combination of isoelectric focusing and gel gradient electrophoresis, the enzyme was separated into two subunits, one having a molecular weight of 70,000, the other less than 20,000. The large subunit has slight enzymatic activiy, while the smaller subunit may be responsible for the charge difference in the holoenzyme pattern. The glycoprotein pattern of the uterine fluid peroxidase is further defined by its separation by affinity chromatography using a concanavalin A-Sepharose column and by its susceptibility to neuraminidase treatment.

Structural study on liver gelactin

1984 ◽  
Vol 62 (11) ◽  
pp. 1072-1075 ◽  
Author(s):  
Julian Gruda ◽  
Hélène-Marie Thérien
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Structural Study ◽  
Isoelectric Focusing ◽  
Isoelectric Point ◽  
Gel Filtration ◽  
Rabbit Liver

Electron microscopy, ultracentrifugation, gel filtration, and isoelectric focusing were carried out with gelactin, an actin-gelling protein from rabbit liver. Gelactin is a dimeric acidic protein (isoelectric point (pI) = 5.45), with a molecular weight of 190 000, a Svedberg constant of 6.25, and a Stoke's radius and length of 7.0 and 28 nm, respectively. While different from α-actinin by pI and amino acid composition, gelactin belongs by its dimensions to the class of α-actinins.

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