Summary
Phenolase was found in all subcellular fractions of the potato tuber roughly in proportion to the protein content of each fraction. Approximately 75% of the total phenolase activity was found in the supernatant after high speed (90,000 x g) centrifugation. Less than 5% of the total activity was associated with the particulate fraction sedimented at either 12,000 x g or 90,000 x g. The phenolase enzyme associated with the particulates sedimented at either 12,000 x g or 90,000 x g was progressively removed by successive washings indicating that the enzyme was only loosely associated with and not a fixed part of the particulates. The phenolase associated with the cell-wall residue and cell debris (sedimented at 600 x g) showed only a small fraction of the total activity. This residue-bound enzyme was very resistant to successive washings. Little or no phenolase was located on the surface of potato cells. The catecholase/cresolase ratios, substrate specificities, and Michaelis constants of the phenolase enzyme associated with each of the different fractions were of the same order of magnitude. The differences found were insufficient to conclude that the phenolase enzymes associated with the different fractions were dissimilar.
Resumen
La fenolasa fue encontrada en todas las fracciones subcelulares del tubérculo de la papa, a groso modo en proporción al contenido protéico de cada fracción.. Aproximadamente en 75% del total de actividad de la fenolasa, fue encontrado en la suspención, después del centrifugado a alta velocidad (90,000 x g). Menos del 5% de la actividad total fue asociada con la fracción peculiar sedimentada a 12,000 x g ó 90,000 x g. La enzima fenolasa, asociada con el sedimento peculiar, a 12,000 x g ó 90,000 x g, fue removida regresivanente, por lavados sucesivos, indicando que la enzima solamente fue ligeramente asociada y no una parte fija del peculiar. La fenolasa asociada con los residuos de las células de las paredes y deshechos de células (sedimenta a 600 x g), mostraron sólo una pequeña fracción de la actividad total. Este eslabón de residuo enzimático, fue muy resistente a los lavados sucesivos. Poca o nada de fenolasa fue localizada en la supreficie de las células de la papa. Los radios de catecolasa/cresolasa, especificidades del substrato y constantes de Michaelis de la enzima fenolasa, asociada con cada una de las fracciones diferentes, fueron del mismo orden de magnitud. Las diferencias encontradas fueron insuficientes para concluir que la enzima fenolasa, asociada con las fracciones diferentes, fueron disimilares.
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Craft, C.C. Localization and activity of phenolase in the potato tuber. American Potato Journal 43, 112–121 (1966). https://doi.org/10.1007/BF02862620
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DOI: https://doi.org/10.1007/BF02862620