[Isolation and partial characterization of phenoloxidase from apples (Malus domestica, var. Anna)].
This study pursued the isolation and partial characterization of the enzyme polyphenoloxidase from apple (Malus domestica Anna variety), grown in the Hermosillo Coast (State of Sonora, Mexico). The effects of pH and temperature as well as its specificity towards substrates, and its behavior under conditions of hydrophobic chromatography, were studied. The enzyme was isolated from a residual powder obtained from ripe apples homogenized with cold acetone. The extract thus prepared was used to characterize the enzyme, and it showed an optimum pH of 5.36 and an optimum temperature of 35 degrees C. The substrate specificity proved to decrease from 4-methyl catechol, chlorogenic acid, catechol, and caffeic acid, to 3,4-dihydroxiphenyl alanine (DOPA). The enzyme resulted to be more thermostable (temperature range: 35 degrees C to 60 degrees C) than the rest of oxidases of plant origin. When the extract was eluted under conditions of hydrophobic chromatography separation, it appeared as a single peak resulting in a 300 fold purification. The phenolase activity characteristics found in the present study were similar to those observed in other apples from temperate climates; however, this particular polyphenoloxidase is more thermostable under natural conditions. This explains why apples of the Anna variety, at the high harvesting temperature, show a very fast formation of brown spots even when there is a minor damage. The content of compounds with phenolic group was high (1.16 g/100 g fresh weight). Further increase of the velocity of fruit enzymatic browning was due to this reason.
Soto-Valdéz
H
H
Centro de Investigación en Alimentación y Desarrollo, A. C. Hermosillo, Sonora, México.
Trejo-González
A
A
D002845:Chromatography; D005638:Fruit / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D014442:Monophenol Monooxygenase / Q000302:isolation & purification*; D013696:Temperature
D014442:Monophenol Monooxygenase
Aislamiento y caracterización parcial de la enzima fenoloxidasa de manzana (Malus domestica, var. Anna).