Kinetic parameters for the thermal inactivation of several enzymes in carrot and potato homogenates have been determined. In carrots the most heat-resistant activity was polygalacturonase, followed by peroxidase and pectinmethylesterase. In potatoes peroxidase was the most resistant, followed by pectin methylesterase, polyphenol oxidase, and lipoxygenase. There were several notable similarities between the inactivation kinetics in the two vegetables. In both cases peroxidase activity gave simple first-order inactivation kinetics but yielded a curved Arrhenius plot for the temperature dependence. Pectin methylesterase in both commodities consisted of a labile and a resistant form. The relative amounts of the two forms and the temperature dependences for their inactivation were also similar.
Vine holding of ripe fruit adversely affects quality, especially pH and titratable acidity. Recent problems with high tomato juice pH encountered by tomato processors in California could be the result of increased average fruit maturity at harvest.
The underlying mechanisms governing nonenzymatic pectin and pectate degradation during thermal treatment have not yet been fully elucidated. This study determined the extent of nonenzymatic degradation due to beta-elimination, acid hydrolysis, and demethylation during prolonged heating of citrus pectins and its influence on physicochemical properties. Solutions of citrus pectins, buffered from pH 4.0 to 8.5, were heated at 75, 85, 95, and 110 degrees C for 0-300 min. Evolution of methanol and formation of reducing groups and unsaturated uronides were monitored during heating. Molecular weight and viscosity changes were determined through size exclusion chromatography and capillary viscometry, respectively. Results showed that at pH 4.5, the activation energies of acid hydrolysis, beta-elimination, and demethylation are 95, 136, and 98 kJ/mol, respectively. This means that at this pH, acid hydrolysis occurs more rapidly than beta-elimination. Furthermore, the rate of acid hydrolysis is diminished by higher levels of methyl esterification. Also, citrus pectin (93% esterified) degrades primarily via beta-elimination even under acidic conditions. Acid hydrolysis and beta-elimination caused significant reduction in relative viscosity and molecular weight.
Three colorimetric reagents for the determination of formaldehyde, the Nash reagent (ammonia plus acetylacetone), Purpald (4-amino-3-hydrazino-5-mercapto-1,2,4-triazole), and N-methylbenzothiazolinone-2-hydrazone (MBTH), were compared for the determination of methanol when used in conjunction with alcohol oxidase. The combination of alcohol oxidase plus the commonly used Nash reagent was specific for methanol versus ethanol, but had the lowest sensitivity of the three reagents tested. Substituting Purpald for the Nash reagent increased the sensitivity 3-fold while still maintaining a high (59-fold) selectivity for methanol versus ethanol. Using MBTH increased the sensitivity still further, but with a loss of the selectivity toward methanol. Since MBTH reacted with aldehydes under neutral conditions, it could be included along with the alcohol oxidase to act as an aldehyde trap. This prevented further oxidation reactions by alcohol oxidase and allowed for extended incubations. A procedure for assaying low levels of pectin methylesterase activity that relies on this trapping ability is described. In addition, alcohol oxidase plus Purpald is shown to be a simple and sensitive way to measure the methanol released from plant material following the thermal activation of endogenous pectin methylesterase.
Modifications to the commonly used Schwimmer and Weston procedure for determining pyruvic acid in onions are proposed. These modifications involve changes in the concentrations and volumes of the reagents and the use of 515 rather than 420 nm as the wavelength for measurements. These changes improve both the linearity and the sensitivity of the assay and eliminate interference from coloured material found in yellow onions. With these changes, more reliable determinations of pyruvic acid and thus of onion pungency should be possible.
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