Variation in Quantity of Methanol Recovered from Raisins

1963 ◽  
Vol 46 (2) ◽  
pp. 341-343
Author(s):  
M Alice Brown ◽  
James R Woodward ◽  
Floyd DeEds

Abstract The amount of naturally occurring methanol in fruit must be known so that the quantity left as fumigation residue can be determined. In a study of methanol content of raisins, which had given inconsistent results, the raisins were subjected to different conditions of treatment immediately prior to methanol determination. Conditions that favored pectin esterase activity gave higher values for methanol content than conditions known to inactivate enzymes. Evidence was also obtained that both chemical and enzymic hydrolysis of methyl ester groups of pectic materials occur during analysis.

1968 ◽  
Vol 19 (03/04) ◽  
pp. 334-345
Author(s):  
E. R Cole

SummaryThe ratio of TAME esterase activity (or PME esterase activity for α-chymotrypsin) to HME esterase activity has been determined for a number of proteolytic enzymes under identical conditions. Papain showed no attack on HME, but ficin, bromelin, human plasmin, trypsin, thrombin, autoprothrombin C, urokinase and α-chymotrypsin hydrolyze this substrate to varying degrees. Ficin showed a TAME/HME activity ratio of 304 while the lowest ratio (approximately 1) was found for urokinase, an activator of plasminogen. This high relative affinity of urokinase for HME prompted an investigation to determine if this property was also a function of streptokinase-activator complex. Glycerol-activated plasmin had a low relative affinity for HME as a substrate (TAME/HME = 157), but addition of high concentrations of streptokinase increased the relative affinity of the preparation for HME catalysis by 500 to 600% (TAME/HME = 26.4). Human plasminogen activated by high concentrations of streptokinase also demonstrate high relative affinity for HME (TAME/ HME = 20-25). HME esterase activity is largely lost when streptokinase is destroyed by pH 2.0 treatment, but can be regained by readdition of streptokinase to the plasmin preparation. Hydrolysis of HME by plasmin preparations in the presence of streptokinase is believed to be primarily a property of streptokinase-plasmin complex.


1959 ◽  
Vol 197 (6) ◽  
pp. 1178-1180 ◽  
Author(s):  
Ricardo H. Landaburu ◽  
Walter H. Seegers

In the activation of purified prothrombin with thrombin, with platelet factor 3, or in 25% sodium citrate solution the free thrombin activity which develops is greater at all times when measured by hydrolysis of p-toluenesulfonyl-arginine-methyl ester than when measured by the clotting of fibrinogen. Since the esterase activity appears before clotting power and remains after clotting power is lost, the clotting property must arise from a dissociable complex arising from the unit that has the esterase function. It is postulated that "clotting thrombin" may be a dimer of the lowest subunit of prothrombin required to have the proteolytic enzyme. The latter could have an important function in our physiology quite apart from the clotting of blood.


1965 ◽  
Vol 43 (12) ◽  
pp. 1961-1970 ◽  
Author(s):  
D. R. Whitaker ◽  
C. Roy ◽  
C. S. Tsai ◽  
L. Jurášek

The proteolytic properties of the α- and β-lytic proteases of a species of Sorangium were compared. Neither enzyme showed evidence of aminopeptidase, carboxypeptidase, or amidase activity in tests with a series of peptides and substituted amino acids at pH 5.2, 7.2, and 9.0. Neither enzyme showed evidence of esterase activity towards N-benzoyl-L-arginine methyl ester at pH 6.8. Hydrolysis of the A chain of oxidized insulin at pH 9 slows down markedly when the α-enzyme has cleaved the chain once; the initial fast cleavage can take place at linkages between residues 9 and 10, 10 and 11, and 12 and 13; more slowly cleaved linkages are between residues 3 and 4, and 8 and 9. Hydrolysis of the B chain by the α-enzyme at pH 9 is still faster and slows down when the chain has been cleaved twice. One fast cleavage is at the linkage between residues 18 and 19; the other can take place at the linkages between residues 12 and 13, and 14 and 15; more slowly cleaved linkages are between residues 8 and 9, 9 and 10, and 15 and 16. Under the conditions tested, the β-enzyme does not hydrolyze the A chain appreciably at pH 9. It cleaves the B chain rapidly at the linkage between residues 23 and 24 and more slowly at linkages between residues 18 and 19. The linkages split by both enzymes are those which involve the carboxyl group of a neutral amino acid.


1968 ◽  
Vol 19 (03/04) ◽  
pp. 321-333
Author(s):  
E. R Cole

SummaryThe hydrolysis of L-histidine methyl ester (HME) by bovine thrombin preparations has been investigated. Activation of purified bovine prothrombin in 25% sodium citrate solution resulted in the simultaneous development of fibrinogen clotting activity and of TAME and HME esterase activities. Most of the HME esterase activity was identified with fibrinogen clotting activity on sequential chromatography of activated prothrombin on DEAE-cellulose and Amberlite CG-50 resin columns, although some HME esterase activity could be demonstrated in concentrates of the autoprothrombin C fraction. The optimum pH for HME hydrolysis by thrombin was found at 7.6 in phosphate and Tris buffered reactions. Tris buffer and other amines depress HME esterase activity of thrombin, while sodium cholate accelerates the reaction. The Michaelis constant, Km, was estimated to be 0.134 M at pH 7.6 in phosphate buffer and at 37° C.


1973 ◽  
Vol 56 (3) ◽  
pp. 361-385 ◽  
Author(s):  
E. D. HELTON ◽  
W. N. HOLMES

SUMMARY After the injection of [1,2-3H]corticosterone into ducks, no unaltered free [1,2-3H]corticosterone was found in either bile, gut contents or cloacal fluid. All the [1,2-3H]corticosterone metabolites in cloacal fluid had similar chromatographic mobilities. A portion of these metabolites was steroid ester sulphate but enzymic hydrolysis suggested that only a small fraction of the sulphate was conjugated to [1,2-3H]corticosterone. The amount of ester sulphate in samples of cloacal fluid, however, was highly variable. After the injection of [1,2-3H]aldosterone one-fifth of the total radioactivity in urine was free [1,2-3H]aldosterone but none was found in the bile and gut contents. Chromatographic purification of the [1,2-3H]aldosterone metabolites in cloacal fluid showed two significant peaks and both peaks contained steroid ester sulphates. Enzymic hydrolysis of the tritiated material showed that each peak also contained a glucuronide of [1,2-3H]aldosterone. The effect of the entero-hepatic circulation on the excretion of tritiated metabolites by the kidney was also examined. Diversion of bile from the duodenum prevented the entry of radioactive material into the gut. The reduced reabsorption of tritiated corticosterone and tritiated aldosterone metabolites from the gut, however, did not affect the amount of tritium excreted by the kidneys. In the case of both hormones, some of the reabsorbed metabolite which had been extracted by the liver was released and re-entered the peripheral blood via the hepatic circulation. The stress of surgery in both the control birds and the birds with bile catheters caused the distribution of [1,2-3H]corticosterone and the tritiated corticosterone metabolites to change; a significantly smaller percentage of the injected radioactivity was recovered from the tissues and excretory fluids. This response to stress was not observed in the [1,2-3H]aldosterone injected birds. When partially purified metabolites of [1,2-2-3H]corticosterone and [1,2-3H]aldosterone were injected into the lumen of the gut, the rate of uptake of the tritiated corticosterone metabolites was higher than that of the tritiated aldosterone metabolites. But, when the metabolites of [1,2-3H]corticosterone and [1,2-3H]aldosterone were injected intravenously, and the rate of the intestinal reabsorption of the metabolites was not limiting, the rates of tritium excretion by the kidneys were the same. The volumes of distribution and metabolic clearance rates of the partially purified [1,2-3H]corticosterone and [1,2-3H]aldosterone metabolites were significantly different from one another and the metabolites could be readily distinguished physiologically as well as chromatographically. The pattern of distribution and the disappearance from plasma of i.v. administered synthetic [1,2-3H]corticosterone-21-SO4 was also distinguishable from the naturally occurring metabolites of [1,2-3H]corticosterone.


1966 ◽  
Vol 16 (01/02) ◽  
pp. 018-031 ◽  
Author(s):  
S Sherry ◽  
Norma Alkjaersig ◽  
A. P Fletcher

SummaryComparative studies have been made of the esterase activity of plasmin and the streptokinase-activator of plasminogen on a variety of substituted arginine and lysine esters. Human plasmin preparations derived by different methods of activation (spontaneous in glycerol, trypsin, streptokinase (SK) and urokinase) are similar in their esterase activity; this suggests that the molecular structure required for such esterase activity is similar for all of these human plasmins. Bovine plasmin, on the other hand, differs from human plasmin in its activity on several of the substrates studied (e.g., the methyl esters of benzoyl arginine and tosyl, acetyl and carbobenzoxy lysine), a finding which supports the view that molecular differences exist between the two animal plasmins. The streptokinase-activator hydrolyzes both arginine and lysine esters but the ratios of hydrolytic activity are distinct from those of plasmin and of other activators of plasminogen. The use of benzoyl arginine methyl ester as a substrate for the measurement of the esterase activity of the streptokinase-activator is described.


2013 ◽  
Vol 69 (11) ◽  
pp. 1225-1228 ◽  
Author(s):  
Sara Wyss ◽  
Irmgard A. Werner ◽  
W. Bernd Schweizer ◽  
Simon M. Ametamey ◽  
Selena Milicevic Sephton

Hydrolysis of the methyl ester (±)-threo-methyl phenidate afforded the free acid in 40% yield,viz.(±)-threo-ritalinic acid, C13H17NO2. Hydrolysis and subsequent crystallization were accomplished at pH values between 5 and 7 to yield colourless prisms which were analysed by X-ray crystallography. Crystals of (±)-threo-ritalinic acid belong to theP21/nspace group and form intermolecular hydrogen bonds. An antiperiplanar disposition of the H atoms of the (HOOC—)CH—CHpygroup (py is pyridine) was found in both the solid (diffraction analysis) and solution state (NMR analysis). It was also determined that (±)-threo-ritalinic acid conforms to the minimization of negativegauche+–gauche−interactions.


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