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.

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. 

Studies on the к-casein complex: II. The isolation of a sialic acid-containing fraction by disrupting the complex at low pH in the presence of sodium chloride

1965 ◽  
Vol 32 (1) ◽  
pp. 57-63 ◽  
Author(s):  
R. Beeby
Keyword(s):  
Sialic Acid ◽  
Sodium Chloride ◽  
Deae Cellulose ◽  
Low Ph ◽  
The Other ◽  
Trichloracetic Acid ◽  
Complex Ii ◽  
Total Sialic Acid

SummaryWhen crude к-casein was precipitated at pH 3 in the presence of 0·4m-NaCl the supernatant contained up to 80% of the total sialic acid but no detectable cystine or cysteine. Two fractions were obtained from this supernatant by chromatography on DEAE cellulose; one containing 4–6% sialic acid and the other only onetenth of this amount.Most of the sialic acid of the sialic acid-rich fraction was soluble in 12% trichloracetic acid following treatment with rennin. It is suggested that the glycopeptide released by the action of the enzyme on casein originates from this fraction.

scholarly journals DEVELOPMENT OF RECOMBINANT PEPTIDE TECHNOLOGY WITH ANTIMICROBIAL PROPERTIES OF A BROAD ACTION SPECTRUM

2017 ◽  
pp. 3-14
Author(s):  
Alexander Prosekov ◽  
Alexander Prosekov ◽  
Olga Babich ◽  
Olga Babich ◽  
Irina Milenteva ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Action Spectrum ◽  
Antimicrobial Properties ◽  
Hplc Method ◽  
Salting Out ◽  
Antimicrobial Spectrum ◽  
Recombinant Peptide ◽  
Isolation And Purification ◽  
Protein Nature

Bacteriocins are antibacterial, mainly complex, substances of protein nature. The promising strains producing bacteriocins used in the food industry are lactic acid microorganisms. This study examines the development of a technology for the production of a recombinant peptide with broad-spectrum antimicrobial properties. An important step is the isolation and purification of the recombinant peptide. It has been proved that the highest antimicrobial activity is manifested by a recombinant peptide isolated by a method based on salting out with ammonium sulfate. During the purification of the recombinant bacteriocin preparation, three kinds of columns were used. In the purification process, the volume of bacteriocin produced decreases 3-fold, while the RU/mL increases 3-fold, and RU/mg increases 6-fold. Purification allows the use of a smaller amount of recombinant bacteriocin in technologies with greater efficacy. Based on the results of determining the molecular weight and purity of the recombinant bacteriocin, it was found that the molecular weight of the recombinant bacteriocin having the amino acid sequence: KYYGNGVTCCKHSCSVDXGKASSCIINNGAMAXATGGH GGNHCCGMSRYIQGIPDFLRGYLHGISSANKHKKGRL, is 13 kDa. A technology for the preparation of a broad-action antimicrobial spectrum peptide has been developed. The process of production of antimicrobial peptide includes such stages as: cultivation of the recombinant strain of Escherichia coli BL21DE3; separation of biomass from the nutrient medium; precipitation of bacteriocins by ammonium sulfate; centrifugation; washing the precipitate; centrifugation at 4200 rpm and separation of the preparation; purification of bacteriocins by HPLC method; packing in bags of polymeric and combined materials; storage at a temperature of 18±2°C for 12 months.

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.

Biosynthesis of chloramphenicol in Streptomyces species 3022a: the nature of the arylamine synthetase system

1979 ◽  
Vol 25 (12) ◽  
pp. 1408-1415 ◽  
Author(s):  
M. M. Francis ◽  
D. W. S. Westlake
Keyword(s):  
Molecular Weight ◽  
Infrared Spectrum ◽  
Pyridoxal Phosphate ◽  
Core Protein ◽  
Deae Cellulose ◽  
The Other ◽  
Small Molecular Weight ◽  
Streptomyces Species ◽  
Effector Molecules ◽  
Chorismic Acid

The arylamine synthetase which catalyses the conversion of chorismic acid to p-aminophenylalanine in Streptomyces species 3022a was separated from aminotransferase by DEAE-cellulose chromatography. Recovered activity was stimulated by the addition of the aminotransferase and pyridoxal phosphate. Activity was further fragmented into three separate components by passage through a Sephadex G-100 column. Only one component produced p-aminophenylalanine but in combination the three stimulated each other's activity. Although the products of the other two components were unstable, an infrared spectrum of one of them was obtained and confirmed the presence of an aromatic amine, but other functional groups could not be ascertained. This product was not recognized as a substrate by the arylamine synthetase complex and it was suggested that it may be a degradation product of an intermediate of p-aminophenylalanine biosynthesis or an unknown intermediate of later biosynthetic steps of the chloramphenicol pathway. It is further suggested that arylamine compounds produced by this organism are the result of interaction of a core protein with other macromolecules and small molecular weight effector molecules.

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.

Two types of xylanases of alkalophilic Bacillus sp. No. C-125

1985 ◽  
Vol 31 (6) ◽  
pp. 538-542 ◽  
Author(s):  
H. Honda ◽  
T. Kudo ◽  
Y. Ikura ◽  
K. Horikoshi
Keyword(s):  
Ammonium Sulfate ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Bacillus Sp ◽  
Ammonium Sulfate Precipitation ◽  
Molecular Weights ◽  
Protein Molecules ◽  
Activity Curve ◽  
Alkalophilic Bacillus ◽  
Hydrolysis Of

One alkalophilic Bacillus sp. strain C-125 (FERM No. 7344) was isolated from soil. From this organism, two types of xylanases, designated xylanase A and xylanase N, were purified by an ammonium sulfate precipitation followed by Biogel P-30 gel filtration, DEAE-cellulose chromatography, and Sephadex G-75 gel filtration. The molecular weights of xylanase A and N were estimated as 43 000 and 16 000, respectively. Immunological experiments indicated that xylanase A and xylanase N were entirely different protein molecules. Xylanase N was most active at pH 6.0–7.0, but xylanase A had a very broad pH activity curve (pH 6–10) and was still active even at pH 12.0. The maximum hydrolysis of xylan by the enzymes was about 25%. Both enzymes split xylan and yielded xylobiose and higher oligosaccharides but could hydrolyze neither xylobiose nor xylotriose. Trans xylosidation activities were detected in both enzymes.

scholarly journals The xylanase system of Coniophora cerebella

1968 ◽  
Vol 108 (4) ◽  
pp. 571-576 ◽  
Author(s):  
N. J. King ◽  
D. B. Fuller
Keyword(s):  
Molecular Weight ◽  
Culture Filtrate ◽  
Uronic Acid ◽  
Enzyme System ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
The Other ◽  
Xylose Residue ◽  
Random Manner ◽  
Acidic Sugars

1. The culture filtrate of the fungus Coniophora cerebella grown on poplar 4-O-methylglucuronoxylan as carbon source and enzyme inducer contained an enzyme system that degraded the polysaccharide to xylose, acidic and neutral oligosaccharides and an enzyme-resistant polymer. Free uronic acid was not produced. 2. Cold ethanol fractionation of the culture filtrate yielded two active fractions, one of which had only xylanase (EC 3.2.1.8) and the other both xylanase and xylosidase (EC 3.2.1.37) activities. Further fractionation on DEAE-cellulose resolved the xylanase and xylosidase activities. 3. The xylanase degraded poplar 4-O-methylglucuronoxylan in an essentially random manner, producing oligosaccharides, but some xylose residues in the vicinity of uronic acid side groups were protected from hydrolysis, preventing a truly random attack. The xylosidase attacked the polysaccharide very slowly, releasing xylose, but the oligosaccharides produced by the action of the xylanase were much more susceptible to hydrolysis by the xylosidase. 4. The products of xylanase action were separated into neutral and acidic fractions. The neutral oligosaccharides were separated by chromatography on charcoal–Celite, and the major products were characterized as xylobiose, xylotriose, xylotetraose and xylopentaose. Some of the acidic sugars were branched, having the uronic acid residue attached to a xylose residue other than the terminal non-reducing one. 5. Gel filtration of various xylanase fractions gave values for the molecular weight of the enzyme from 34000 to 38000.

ADRENAL PHOSPHORYLASE AND THE ACTION OF CORTICOTROPHIN

1960 ◽  
Vol XXXIV (I) ◽  
pp. 69-76 ◽  
Author(s):  
Rolf Emberland
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulfate ◽  
Adenosine Monophosphate ◽  
The Other ◽  
Ammonium Sulfate Precipitation ◽  
Phosphorylase Activity ◽  
Ammonium Sulfate Fractionation ◽  
Sulfate Fractionation

ABSTRACT Corticotrophin preparations are capable of stimulating the phosphorylase activity of fresh beef adrenal extract. The effect disappears when the extract is heated or purified and decreases with increasing age of the extract. 5-adenosine monophosphate has no influence on the phosphorylase activity of fresh beef adrenal extract but has a stimulating effect when the preparation is dialyzed. By ammonium sulfate precipitation, calcium phosphate gel adsorption and ammonium sulfate fractionation, two phosphorylase fractions were obtained, one being influenced by 5-adenosine monophosphate to a much higher degree than the other one. None were influenced by corticotrophin.

scholarly journals The multiple forms of α-D-mannosidase in human plasma

1979 ◽  
Vol 179 (3) ◽  
pp. 583-592 ◽  
Author(s):  
S Hirani ◽  
B Winchester
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Concanavalin A ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
The Other ◽  
Multiple Forms ◽  
Simple Correlation ◽  
Multiple Components

The acidic alpha-D-mannosidase in human plasma closely resembles liver acidic alpha-D-mannosidase in its affinity for concanavalin A-Sepharose, molecular weight and resolution into multiple components on DEAE-cellulose. A combination of chromatography on concanavalin A-Sepharose and gel filtration on Sephadex G-200 and Sepharose 6B suggests that four forms of intermediate alpha-D-mannosidase, which differ either in their molecular weight of affinity for concanavalin A, exist in human plasma. A practical classification and nomenclature for the multiple forms of intermediate alpha-D-mannosidase in plasma based on molecular weight and affinity for concanavalin A is proposed. Multiple forms of intermediate alpha-D-mannosidase were also observed by ion-exchange chromatography on DEAE-cellulose, but there was not a simple correlation between these forms and those obtained with the other separation procedures. The form of intermediate alpha-D-mannosidase least abundant in plasma, approx. 7% of the activity, has very similar properties to the neutral alpha-D-mannosidase in human liver. In contrast, the other three forms of intermediate alpha-D-mannosidase, which account for over 90% of the activity, do not appear to be present in liver, except perhaps in trace amounts.

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