Effect of pH and temperature on peroxidase and polyphenoloxidase activities of litchi pericarp

Abstract

After harvesting litchi, the red color of the fruit pericarp is rapidly lost resulting in discoloration and browning during storage and marketing. The loss of the red color is caused by the degradation or loss of stability of anthocyanins. The action of peroxidase and polyphenoloxidase is usually related to the browning and discoloration of fruits of various species. This study aimed to evaluate the influence of pH and temperature on peroxidase and polyphenoloxidase activities, in a partially purified preparation of pericarp of the litchi cultivar Bengal. Fruits were harvested at the ripe stage and polyphenoloxidase was partially purified by sequential saturation in 80% ammonium sulfate. At concentrations of 40-50% and 60-70% ammonium sulfate the activities of polyphenoloxidase and peroxidase were, respectively, 124 times and 158 times higher than in the crude extract. The activity of peroxidase and polyphenoloxidase was maximum at pH 6.5 and 7.0, respectively, and no activity was detected at pH 2.5 and 9.5. Pre-incubation of the enzyme extract for 45 min at pH 2.5 or 9.5 completely inactivated the enzymes, with the highest degree of efficiency at pH 2.5. Peroxidase activity was highest at 70°C and remained active for a period of 120 min at 70 and 80 o C. Peroxidase became completely inactive when maintained at 90 o C for 10 min or 1 min at 100 o C. Polyphenoloxidase activity was highest at 20°C and remained active for a period of 120 min at 40 and 50 o C and was inactivated after 10 min at 60ºC. Due to the high temperature of inactivation of the peroxidase and polyphenoloxidase activities, the enzymes can be inactivated more easily in fruits using acid or alkaline solutions. Key words: Litchi Chinensis Sonn., enzyme purification, skin browning Efeito do pH e da temperatura nas atividades da peroxidase e polifenoloxidase do pericarpo de lichia RESUMO: Após a colheita do fruto, a cor vermelha do pericarpo da lichia é rapidamente perdida, o que resulta em descoloração e escurecimento durante o armazenamento e comercialização. A perda da cor vermelha é devido à degradação de antocianinas ou à perda de sua estabilidade. Usualmente, a ação da peroxidase e polifenoloxidase está relacionada ao escurecimento e à descoloração de várias frutas. Avaliou-se a influência do pH e da temperatura na atividade da peroxidase e polifenoloxidase em uma preparação purificada parcial de pericarpo de cultivar Bengal. Os frutos foram colhidos no estádio vermelho maduro. A polifenoloxidase foi parcialmente purificada por saturação seqüencial até 80% de sulfato de amônio. Na concentração de 40-50% e de 60-70% de sulfato de amônio, a atividade da polifenoloxidase e peroxidase foi 124 e 158 vezes maior vezes maior do que a encontrada no extrato cru. A peroxidase e polifenoloxidase apresentaram ótima atividade em pH 6,5 e 7,0 e nenhuma atividade foi detectada a pH 2,5 e 9,5. A pré-incubação do extrato das enzimas até 45 min a pH 2,5 ou 9,5 inativou completamente as enzimas, sendo que o maior grau de eficiência ocorreu em pH 2,5. A...

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