Purification and properties of a β-hexosidase from Sporobolomyces singularis

1969 ◽  
Vol 47 (11) ◽  
pp. 1021-1025 ◽  
Author(s):  
J. A. Blakely ◽  
S. L. MacKenzie
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Sedimentation Coefficient ◽  
Polyacrylamide Gel ◽  
Ammonium Sulfate Precipitation ◽  
Purification And Properties

A β-hexosidase has been isolated from Sporobolomyces singularis by conventional techniques involving ammonium sulfate precipitation and chromatography on columns of Sephadex G-200 and DEAE-Sephadex A-50. Electrophoresis on polyacrylamide gel was used as the final preparative step. The sedimentation coefficient (s°20,w) of the enzyme was 7.6 and its molecular weight was in the range 140 000–145 000. Although the β-hexosidase performed the functions of a β-D-galactoside galactohydrolase (β-galactosidase), it also catalyzed the hydrolytic function normally performed by a β-D-glucoside glucohydrolase; both these functions appear to reside in the same molecule.

Extraction and Partial Purification of Protease from Bilimbi (Averrhoa bilimbi L.)

2013 ◽  
Vol 10 (2) ◽  
pp. 29
Author(s):  
Normah Ismail ◽  
Nur' Ain Mohamad Kharoe
Keyword(s):  
Molecular Weight ◽  
Protein Content ◽  
Ammonium Sulfate ◽  
Molecular Weight Distribution ◽  
Weight Distribution ◽  
Protease Activity ◽  
Protein Concentration ◽  
Temperature Stability ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation

Unripe and ripe bilimbi (Averrhoa bilimbi L.) were ground and the extracted juices were partially purified by ammonium sulfate precipitation at the concentrations of 40 and 60% (w/v). The collected proteases were analysed for pH, temperature stability, storage stability, molecular weight distribution, protein concentration and protein content. Protein content of bilimbi fruit was 0.89 g. Protease activity of both the unripe and ripe fruit were optimum at pH 4 and 40°C when the juice were purified at 40 and 60% ammonium sulfate precipitation. A decreased in protease activity was observed during the seven days of storage at 4°C. Molecular weight distribution indicated that the proteases protein bands fall between IO to 220 kDa. Protein bands were observed at 25, 50 and 160 kDa in both the unripe and ripe bilimbi proteases purified with 40% ammonium sulfate, however, the bands were more intense in those from unripe bilimbi. No protein bands were seen in proteases purified with 60% ammonium sulfate. Protein concentration was higher for proteases extracted with 40% ammonium sulfate at both ripening stages. Thus, purification using 40% ammonium sulfate precipitation could be a successful method to partially purify proteases from bilimbi especially from the unripe stage. 

scholarly journals PURIFICATION AND PHYSICAL PROPERTIES OF GROUP C STREPTOCOCCAL PHAGE-ASSOCIATED LYSIN

1971 ◽  
Vol 133 (5) ◽  
pp. 1105-1117 ◽  
Author(s):  
Vincent A. Fischetti ◽  
Emil C. Gotschlich ◽  
Alan W. Bernheimer
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Physical Properties ◽  
Total Protein ◽  
Sedimentation Coefficient ◽  
Polyacrylamide Gel ◽  
Single Band ◽  
Purification Procedure ◽  
Sh Group ◽  
Stokes Radius

A purification procedure for the Group C phage-associated lysin is described utilizing tetrathionate to protect the enzyme's -SH group(s) from thiol-inactivating agents. A 652-fold purification has been accomplished yielding a solution in which the enzyme activity corresponds to essentially a single band on polyacrylamide gel which accounts for 70% of the total protein in the preparation. A molecular weight of 101,000 and frictional ratio of 1.526 was determined for the lysin utilizing experimentally determined values for its Stokes radius and sedimentation coefficient.

Acid Phosphatases Associated With Phosphorus Deficiency in Wheat: Partial Purification and Properties

1991 ◽  
Vol 18 (6) ◽  
pp. 615 ◽  
Author(s):  
RE Guthrie ◽  
KD Mclachlan ◽  
DGD Marco
Keyword(s):  
Molecular Weight ◽  
Phosphorus Deficiency ◽  
Size Exclusion ◽  
Partial Purification ◽  
Acid Phosphatases ◽  
Ammonium Sulfate Precipitation ◽  
Con A ◽  
Substrate Utilisation ◽  
Purification And Properties ◽  
Exclusion Chromatography

Procedures for the partial purification of two phosphatase isozymes found in phosphorus deficient wheat plants are given. The method employs ammonium sulfate precipitation and hydroxylapatite, Con A-Sepharose, anion exchange and size exclusion chromatography. Measurements of their Km, Vmax, pI and molecular weight are reported. Evidence is provided that there are empirical differences in substrate utilisation between these phosphatase isozymes associated with phosphorus deficient plants.

Extraction and Partial Purification of Protease from Bilimbi (Averrhoa bilimbi L.)

2013 ◽  
Vol 10 (2) ◽  
pp. 30
Author(s):  
Normah Ismail ◽  
Nur' Ain Mohamad Kharoe
Keyword(s):  
Molecular Weight ◽  
Protein Content ◽  
Ammonium Sulfate ◽  
Molecular Weight Distribution ◽  
Weight Distribution ◽  
Protease Activity ◽  
Protein Concentration ◽  
Temperature Stability ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation

Unripe and ripe bilimbi (Averrhoa bilimbi L.) were ground and the extracted juices were partially purified by ammonium sulfate precipitation at the concentrations of 40 and 60% (w/v). The collected proteases were analysed for pH, temperature stability, storage stability, molecular weight distribution, protein concentration and protein content. Protein content of bilimbi fruit was 0.89 g. Protease activity ofboth the unripe and ripe fruit were optimum at pH 4 and 40°C when the juice were purified at 40 and 60% ammonium sulfate precipitation. A decreased in protease activity was observed during the seven days of storage at 4°C. Molecular weight distribution indicated that the proteases protein bands fall between IO to 220 kDa. Protein bands were observed at 25, 50 and 160 kDa in both the unripe and ripe bilimbi proteases purified with 40% ammonium sulfate, however, the bands were more intense in those from unripe bilimbi. No protein bands were seen in proteases purified with 60% ammonium sulfate. Protein concentration was higher for proteases extracted with 40% ammonium sulfate at both ripening stages. Thus, purification using 40% ammonium sulfate precipitation could be a successful method to partially purify proteases from bilimbi especially from the unripe stage.

A GENERAL METHOD FOR THE ISOLATION AND PURIFICATION OF PHOSVITIN FROM VERTEBRATE EGGS

1966 ◽  
Vol 44 (12) ◽  
pp. 1647-1655 ◽  
Author(s):  
R. A. Wallace ◽  
D. W. Jared ◽  
A. Z. Eisen
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Deae Cellulose ◽  
The Other ◽  
Chemical Analyses ◽  
Ammonium Sulfate Precipitation ◽  
Isolation And Purification ◽  
Complex Ii ◽  
Vertebrate Eggs ◽  
General Method

A general method has been developed for the isolation and purification of phosvitin from vertebrate eggs. The method is detailed in three steps consisting of (i) isolation of a phosvitin–lipovitellin complex; (ii) ammonium sulfate precipitation of the lipovitellin; and (iii) DEAE-cellulose chromatography of the remaining phosvitin. Phosvitin was isolated from the eggs of five representative vertebrates (lamprey, trout, frog, turtle, and chicken), and chemical analyses together with sedimentation studies were performed on the samples. Preparations were obtained with the lowest N/P ratios reported to date. The analytical results also suggested that trout phosvitin has approximately half the molecular weight (24,000) of the other proteins and that "purified" phosvitin may still be heterogeneous.

scholarly journals The purification and properties of isocitrate lyase from Chlorella

1967 ◽  
Vol 105 (1) ◽  
pp. 409-416 ◽  
Author(s):  
P. C. L. John ◽  
P. J. Syrett
Keyword(s):  
Molecular Weight ◽  
Diffusion Coefficient ◽  
Gel Electrophoresis ◽  
Isocitrate Lyase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sedimentation Coefficient ◽  
Chlorella Pyrenoidosa ◽  
Turnover Number ◽  
Purification And Properties ◽  
Stokes Radius

1. Isocitrate lyase (threo-ds-isocitrate glyoxylate-lyase, EC 4.1.3.1) has been purified from acetate-adapted cells of Chlorella pyrenoidosa. 2. The final preparation was homogeneous by the criteria of sedimentation, diffusion and polyacrylamide-gel electrophoresis. 3. The sedimentation coefficient (S20,w) was 9·04×10−13sec. and the diffusion coefficient (D20,w) 4·62×10−7cm.2/sec.; from these values the molecular weight of the enzyme was calculated to be 170000 and its Stokes radius to be 4·63×10−7cm. 4. The elution of the enzyme from Sephadex G-100 was studied and estimates of molecular weight and Stokes radius were obtained from the elution data. 5. The turnover number of the enzyme was 5950moles of glyoxylate formed/min./mole of enzyme at 30°. 6. With threo-ds(+)-isocitrate as substrate, the Km of the enzyme was 0·023mm.

scholarly journals Purification and properties of oestrogen-induced uterine peroxidase

1975 ◽  
Vol 151 (2) ◽  
pp. 275-279 ◽  
Author(s):  
T McNabb ◽  
P H Jellinck
Keyword(s):  
Molecular Weight ◽  
Peroxidase Activity ◽  
Half Life ◽  
Polyacrylamide Gel ◽  
Gel Chromatography ◽  
Eosinophil Peroxidase ◽  
Purification And Properties

1. An enzyme that can be induced in rat uteri by oestrogens and that catalyses the oxidation of guaiacol and the metabolism and binding of [4-14C]oestradiol to protein in the presence of H2O2 was partially purified by (NH4)2SO4 fractionation and polyacrylamide-gel chromatography. 2. The molecular weight of this uterine peroxidase was estimated to be about 40 000 and thus shown to differ from that of eosinophil peroxidase. 3. Cycloheximide, which blocks the increase in peroxidase activity brought about by oestrogen, was used to determine the half-life (about 4h) of the induced uterine enzyme.

scholarly journals Purification and properties of pantohenase from Pseudomonas fluorescens

1976 ◽  
Vol 157 (2) ◽  
pp. 409-413 ◽  
Author(s):  
R K Airas ◽  
E A Hietanen ◽  
V T Nurmikko
Keyword(s):  
Molecular Weight ◽  
Pseudomonas Fluorescens ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Purification Procedure ◽  
Subunit Molecular Weight ◽  
Purification And Properties

Pantothenase (EC 3.5.1.22) from Pseudomonas fluorescens UK-1 was purified to homogeneity as judged by disc-gel electrophoresis and isoelectric focusing. The purification procedure consisted of four steps: DEAE-Sephadex chromatography, (NH4)2SO4 precipitation, hydroxyapatite chromatography and preparative polyacrylamide-gel electrophoresis. Gel filtration on Ultrogel AcA 34 was used to determine the molecular weight, and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis to study the subunit molecular weight. The enzyme appeared to be composed of two subunits with mol.wts. of approx. 50000 each. The total mol.wt. of the enzyme was thus about 100000. The isoelectric point was 4.7 at 10 degrees C.

A Bacteriocin Produced by Lactobacillus brevis KLDS1.0373 Isolated from “Jiaoke”, a Traditional Fermented Cream from China

2011 ◽  
Vol 183-185 ◽  
pp. 1132-1136
Author(s):  
Hui Wang ◽  
Han Sheng Gong ◽  
Xiang Chen Meng ◽  
Li Li Man
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Inner Mongolia ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Lactobacillus Brevis ◽  
Ammonium Sulfate Precipitation ◽  
Gel Filtration Chromatography

A bacteriocin produced by Lactobacillus brevis KLDS1.0373 which was isolated from “Jiaoke”, a traditional, naturally fermented cream from Inner Mongolia in China was reported in this article. The bacteriocin was partially purified by ammonium sulfate precipitation followed by sequential gel filtration chromatography, and the apparent molecular weight of the partially purified bacteriocin was estimated at approximately 3.8 kDa.

scholarly journals Proteinases from buckwheat (Fagopyrum esculentum moench) seeds: Purification and properties of the 47 kDa enzyme

2006 ◽  
Vol 58 (3) ◽  
pp. 171-177 ◽  
Author(s):  
Gordana Timotijevic ◽  
Svetlana Radovic ◽  
Vesna Maksimovic
Keyword(s):  
Affinity Chromatography ◽  
Ammonium Sulfate ◽  
Proteolytic Activity ◽  
Membrane Fraction ◽  
Aspartic Proteinase ◽  
Ammonium Sulfate Precipitation ◽  
Milk Clotting ◽  
Purification And Properties ◽  
Fagopyrum Esculentum Moench ◽  
Pepstatin A

Aspartic proteinases from buckwheat seeds are analyzed. Three forms of 47 kDa, 40 kDa and 28 kDa, were purified from mature buckwheat seeds, while two forms of 47 kDa and 28 kDa were detected in developing buckwheat seeds using pepstatin A affinity chromatography. A form of 47 kDa was selectively precipitated from other forms by ammonium sulfate precipitation. This enzyme resembles the chymosin-like pattern of proteolytic activity, as it was shown using BSA and k-casein as substrates, clarifying its ability for milk-clotting. The 47 kDa aspartic proteinase form is localized in the membrane fraction. .

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