Investigation of the globulins of cotton seeds VIII. Chymotryptic hydrolysis of the 7S-globulin. Isolation and purification of a chymotryptic peptide

1977 ◽  
Vol 13 (4) ◽  
pp. 467-471
Author(s):  
N. L. Ovchinnikova ◽  
M. A. Kuchenkova ◽  
P. Kh. Yuldashev
Keyword(s):  
Isolation And Purification ◽  
Chymotryptic Peptide ◽  
7S Globulin ◽  
Cotton Seeds ◽  
Hydrolysis Of

scholarly journals UNUSUAL WAY OF REACTION OF 3-AMINO-4-(5-CHLOROMETHYL-1,2,4-OXADIAZOLE-3-YL)-FURAZAN WITH HYDRAZINE

2017 ◽  
Vol 60 (4) ◽  
pp. 26
Author(s):  
Elena V. Stepanova ◽  
Andrei I. Stepanov
Keyword(s):  
Carboxylic Acid ◽  
Catalytic Amount ◽  
Acid Synthesis ◽  
Chloromethyl Group ◽  
One Pot ◽  
Isolation And Purification ◽  
Long Duration ◽  
Reaction Proceeds ◽  
Selective Reactivity ◽  
Hydrolysis Of

The results of our study of the pathways of selective reactivity of 3-amino-4-(5-chloromethyl-1,2,4-oxadiazole-3-yl)furazan versus 5-unsubstituted or 5-methyl and 5-trifluoromethyl substituted 4-(5R-1,2,4-oxadiazole-3-yl)furazans (R = H, Me, CF3) towards the action of hydrazine are discussed. If the reductive opening of 1,2,4-oxadiazole ring in unsubstituted at the С-5 atom (1,2,4-oxadiazol-3-yl)furazan derivatives under the treatment with hydrazine can be used as a method for the preparation of a range of amidrazones of 4-R-furazan-3-carboxylic acid. 3-amino-4-(5-trifluoromethyl-1,2,4-oxadiazol-3-yl)furazan with hydrazine gives amidoxime of 4-aminofurazan-3-carboxylic acid. 3-amino-4-(5-methyl-1,2,4-oxadiazol-3-yl) furazan is inert to the action of hydrazine, on the contrary the reaction of 3-amino-4-(5-chloromethyl-1,2,4-oxadiazole-3-yl)furazan with hydrazine leads to oxidation of chloromethyl group of titled compound to the carbonyl one. In this case the product of reaction of 3-amino-4-(5-chloromethyl-1,2,4-oxadiazole-3-yl)furazan with hydrazine was isolated in a form of corresponding hydrazonomethyl derivative notably as 3-amino-4-(5-hydrazonomethyl-1,2,4-oxadiazole-3-yl)furazan. A possible reaction mechanism for the formation of hydrazonomethyl group by oxidation reaction of chloromethyl group by hydrazine is proposed. 3-Amino-4-(5-hydrazonomethyl-1,2,4-oxadiazol-3-yl)furazan undergoes a transhydrazination reaction with semicarbazide and thiosemicarbazide. But our attempts to its hydrolysis for the purpose to obtain free aldehyde were unsuccessful. Thus, hydrolysis of hydrazonomethyl derivative in acetic acid in the presence of catalytic amount of sulfuric acid results in azine – N,N'-bis(3-(4-aminofurazan-3-yl)-1,2,4-oxadiazol-5-ylmethylyden)hydrazine – precipitation, long-duration boiling in hydrochloric acid leads to Kishner-Wolff reduction of the carbonyl group to 3-amino-4-(5-methyl-1,2,4-oxadiazol-3-yl)furazan, and hydrolysis in alkaline medium leads to 1,2,4-oxadiazole ring opening to amidoxime of 4-aminofurazan-3-carboxylic acid. Synthesis of 3-amino-4-(5-chloromethyl-1,2,4-oxadiazole-3-yl)furazan (R = CH2Cl) was carried out by condensation of amidoxime of 4-aminofurazan-3-carboxylic acid with an excess of chloroacetyl chloride in toluene at elevated temperature. The reaction proceeds through formation of intermediate product – 3-chloromethylamino-4-(5-chloromethyl-1,2,4-oxadiazol-3-yl)furazan. Removing of N-chloroacetyl group in such obtained intermediate was performed by hydrolysis in acidic media. One-pot synthesis without the need for isolation and purification of intermediate is allowed. The structures of obtained compounds were proved by modern methods of physical-chemical analysis (1H, 13C NMR, IR and MS spectroscopy).Forcitation:Stepanova E.V., Stepanov A.I. Unusual way of reaction of 3-amino-4-(5-chloromethyl-1,2,4-oxadiazole-3-yl)furazan with hydrazine. Izv. Vyssh. Uchebn. Zaved. Khim. Khim. Tekhnol. 2017. V. 60. N 4. P. 26-32.      

The globulins of cotton seeds X. Structures of tryptic peptides of the 7S globulin

1977 ◽  
Vol 13 (5) ◽  
pp. 563-566 ◽  
Author(s):  
É. F. Redina ◽  
M. A. Kuchenkova ◽  
L. G. Petrosyan ◽  
P. Kh. Yuldashev
Keyword(s):  
Tryptic Peptides ◽  
7S Globulin ◽  
Cotton Seeds

THE EXTRACELLULAR POLYSACCHARIDE OF GELATINOUS STRAINS OF CHROMOBACTERIUM VIOLACEUM

1960 ◽  
Vol 6 (2) ◽  
pp. 153-163 ◽  
Author(s):  
William A. Corpe
Keyword(s):  
Uronic Acid ◽  
Extracellular Polysaccharide ◽  
Amino Sugar ◽  
Water Soluble ◽  
Chromobacterium Violaceum ◽  
Isolation And Identification ◽  
Isolation And Purification ◽  
Viscous Solution ◽  
Hydrolysis Of

A method for the isolation and purification of the voluminous extracellular polysaccharide of Chromobacterium violaceum is presented. The purified product was fibrous when wet but dried into a tough, pliable film which was completely water soluble, forming a highly viscous solution. Hydrolysis of the polysaccharide, isolation and identification of the components established the presence of glucose as the principal sugar. A uronic acid and an amino sugar not conclusively identified were also present. Glucose, uronic acid, and the amino sugar were found in an approximate 5:1:1 ratio.

A study of the globulins of cotton seeds VI. Peptides of a trytpic hydrolyzate of subunit I of the 7S globulin

1977 ◽  
Vol 13 (4) ◽  
pp. 493-494
Author(s):  
E. F. Redina ◽  
M. A. Kuchenkova ◽  
P. Kh. Yuldashev
Keyword(s):  
7S Globulin ◽  
Cotton Seeds

Investigation of the globulins of cotton seeds VII. Peptides of a triptic hydrolyzate of the 7S-globulin

1977 ◽  
Vol 13 (4) ◽  
pp. 464-467
Author(s):  
É. F. Redina ◽  
M. A. Kuchenkova ◽  
P. Kh. Yuldashev
Keyword(s):  
7S Globulin ◽  
Cotton Seeds

The globulins of cotton seeds XI. The structure of the chymotryptic peptides of the 7S globulin

1977 ◽  
Vol 13 (5) ◽  
pp. 566-569 ◽  
Author(s):  
N. L. Ovchinnikova ◽  
M. A. Kuchenkova ◽  
T. D. Kasymova ◽  
P. Kh. Yuldashev
Keyword(s):  
7S Globulin ◽  
Cotton Seeds

Comparative study of methods for the isolation and purification of bovine κ-casein and its hydrolysis by chymosin

1996 ◽  
Vol 63 (1) ◽  
pp. 61-71 ◽  
Author(s):  
Kate P. Coolbear ◽  
David F. Elgar ◽  
Tim Coolbear ◽  
John S. Ayers
Keyword(s):  
Chemical Composition ◽  
Gel Electrophoresis ◽  
Reversed Phase ◽  
Reversed Phase Chromatography ◽  
Isolation And Purification ◽  
N Terminus ◽  
Hydrolysis Of ◽  
Single Batch ◽  
Made In ◽  
Final Purification

Summaryκ-Casein was purified from a single batch of whole acid casein (κ-A variant) using different methods in order to compare their merits in producing a purified material with a carbohydrate and phosphate heterogeneity representative of the whole κ-casein complement in milk. Ion-exchange methods of purification gave products of higher purity than precipitation techniques involving final purification by ethanol fractionation, but all methods resulted in κ-caseins of apparently similar heterogeneity and chemical composition. The purified κ-caseins were hydrolysed with chymosin and the derived macropeptides isolated. These were all virtually identical as determined by reversed-phase chromatography and gel electrophoresis. Some observations on chymosin hydrolysis of κ-casein were made. In addition to formation of the major para-κ-casein (Glu1–Phe105) and macropeptide (Met106–Val169), chymosin hydrolysis at pH 6·6 also resulted in two minor para-κ-caseins with N-termini corresponding to Phe18 and Ser33 of κ-casein. At pH 5·5 and 4·5 para-κ-casein was rapidly hydrolysed into at least six fragments, one of which had an N-terminus corresponding to Trp76 of κ-casein. At pH 6·6, 5·5 and 4·5 the κ-casein macropeptide was stable to chymosin, but at pH 2·3 it was hydrolysed by chymosin into fragments with N-termini corresponding to Met106, He125, Ala138, Val139, Thr145 and Glu147 of κ-casein.

scholarly journals Improvement of the method of obtaining human IgA Fc-fragments

2015 ◽  
Vol 6 (1) ◽  
pp. 32-35
Author(s):  
O. Y. Galkin ◽  
Y. V. Gorshunov ◽  
V. F. Solovjova
Keyword(s):  
Enzymatic Hydrolysis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Inhibitory Effect ◽  
Nitrogen Atmosphere ◽  
Maximum Output ◽  
Serological Tests ◽  
Isolation And Purification ◽  
Antibody Conjugate ◽  
Hydrolysis Of

To address a number of fundamental and applied problems in immunology, molecular and cellular biology and biotechnology it is necessary to obtain Fc-fragments of immunoglobulins. Fc-fragments may be used for studying of the effector functions of antibodies which are mediated by these areas. They are often used as an immunogen to produce anti-specie (based on so-called secondary antibody) conjugate in the development of serological tests for diagnostics (predominantly such conjugate based on monoclonal antibodies). The work is aimed to develop improved methods of obtaining and allocation of Fc-fragments of human IgA. To achieve this objective, optimization of hydrolysis of IgA with subsequent purification of Fс-fragments have been carried out. Improved method of obtaining Fc-fragments of IgA provides: papain hydrolysis of immunoglobulin in the environment of nitrogen for 4 h, allowing to achieve maximum output of Fc-fragments without their further degradation: isolation and purification of Fc-fragments of human IgA by one-stage gel filtration on sephadex G-100; control of purity of the target product by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate and Ouchterlony immunodiffusion. Enzymatic hydrolysis was carried out at the optimal temperature of papain (37 °C). As the oxygen in the air may have inhibitory effect on enzymatic hydrolysis reaction, the reaction mixture was incubated in the nitrogen atmosphere to prevent inactivation of papain. To reduce the incident degradation of immunoglobulin molecules, papain hydrolysis was carried out without using an enzyme activator (cysteine). Usage of the proposed scheme allows obtaining Fc-fragments of human IgA of high purity. Outcome of Fc-fragments after all stages of purification was about 18% of the initial amount of IgA in the preparation. Molecular weight from Fc-fragments of human IgA was equal to approximately 70 kDa.

Signal resistance of a soluble protein to enzymic proteolysis. An unorthodox approach to the isolation and purification of germin, a rare growth-related protein

1984 ◽  
Vol 62 (12) ◽  
pp. 1351-1353 ◽  
Author(s):  
Z. F. Grzelczak ◽  
B. G. Lane
Keyword(s):  
Soluble Protein ◽  
Sodium Dodecyl ◽  
Related Protein ◽  
Nucleosome Assembly ◽  
Polymeric Form ◽  
Isolation And Purification ◽  
Assembly Factor ◽  
Gastric Pepsin ◽  
Wheat Embryos ◽  
Hydrolysis Of

Onset of growth in germinating wheat embryos is marked by the conspicuous synthesis of germin, a soluble homopentameric protein. Germin is unusually stable in reducing environments containing sodium dodecyl sulfate, but the polymeric form is converted to a protomer (ca. 26 kdaltons) by brief heat treatment. In respect to these physical properties, germin is similar to nucleoplasmin, the putative nucleosome-assembly factor in Xenopus oocytes. To expand the comparison, we treated germin with gastric pepsin in the expectation that pepsin-catalyzed hydrolysis of germin might generate a series of fragments of the kind derived by pepsin digestion of nucleoplasmin. To our surprise, germin was refractory under conditions used to degrade nucleoplasmin. Further study has shown that germin exhibits a measure of stability toward the action of broad-specificity proteases which is unprecedented for a soluble protein. In this report, we document the remarkable resistance of this growth-related protein to enzymic proteolysis and project how this property may make it possible to isolate and purify an otherwise intractably rare, but "interesting" protein.

scholarly journals STUDY OF SOME ENZYME SYSTEMS BY MEANS OF THE DIFFERENTIAL MICRORESPIROMETER

1951 ◽  
Vol 34 (4) ◽  
pp. 431-437
Author(s):  
Nathan Entner ◽  
Paul L. Kirk
Keyword(s):  
Yeast Cells ◽  
Acetyl Phosphate ◽  
Isolation And Purification ◽  
Enzyme Systems ◽  
Enzyme Isolation ◽  
Dehydrogenase System ◽  
Clostridium Kluyveri ◽  
Hydrolysis Of

Three enzymic phenomena, inactivation by oxygen, QOO2 measurements of a dehydrogenase system, and hydrolysis of acetyl phosphate by a phosphatase of Clostridium kluyveri, were studied by means of the differential microrespirometer. Respiration of yeast cells was also measured with the same instrument. All results obtained in the four types of study agreed closely with earlier results obtained with the Warburg apparatus. The amount of sample needed was of the order of 1/100 to 1/200 of that necessary for comparable study with the Warburg apparatus. The advantages of the instrument in enzyme isolation and purification studies are discussed.

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