三疣梭子蟹酚氧化酶的生化特性研究.pdf

172 2010, Vol. 31, No. 01 食品科学 ※生物工程 1 2 , 1 1 1 1 郑 丹 ，段青源 * ，朱励华 ，钟惠英 ，杨家锋 ，汪 杰 (1.宁波市渔业环境与产品质量检验监测中心，浙江宁波 315012 ；2.宁波市海洋与渔业研究院，浙江宁波 315012) ：对三疣梭子蟹酚氧化酶的生化特性进行研究。结果表明：以邻苯二酚为底物，酚氧化酶在pH7.8 ～8.4 时最稳定，最适pH 值为8.2 。温度55 ℃时，酚氧化酶活力最大，在25 ～40 ℃时，热稳定性最好。酶促褐变反 应动力学符合米氏方程，温度为30 ℃，pH7.2 时，米氏常数Km=0.531mol/L ，最大反应速率0.049 A 530nm/min 。选 用的12 种抑制剂对三疣梭子蟹酚氧化酶均有一定的抑制效果，以植酸抑制效果最强，焦亚硫酸钠、L - 半胱氨酸、 草酸次之，抗坏血酸效果较好。 ：三疣梭子蟹；酚氧化酶；褐变 Biochemical Characteristics of Phenoloxidase from Portunus trituberculatus 1 2, 1 1 1 1 ZHENG Dan ，DUAN Qing-yuan *，ZHU Li-hua ，ZHONG Hui-ying ，YANG Jia-feng ，WANG Jie (1. Ningbo Inspection and Monitor Centre for Product Quality and Fishery Environment, Ningbo 315012, China ； 2. Ningbo Academy of Ocean and Fishery, Ningbo 315012, China) Abstract ：Crude phenoloxidase (PO) was extracted from Portunus trituberculatus and characterized biochemically. This enzyme exhibited an optimal pH of 8.2 and temperature of 55 ℃ for catalyzing the oxidation of pyrocatechol as a substrate. Optimal enzyme stability was observed in the ranges of pH 7.8 －8.4 and 25－40 ℃. Browning reaction catalyzed by this enzyme obeyed the Michaelis-Menten equation. The values of Km and Vmax at 30 ℃ and pH 7.2 were calculated to be 0.531 mol/L and 0.049 A530nm/min, respectively. Twelve inhibitors tested all had a certain inhibitory effect against this enzyme, among which phytic acid was the strongest inhibitor, followed in turn by sodium metabisulfite, L-cysteine, oxalic acid and ascorbic acid. Key words ：Portunus trituberculatus ；phenoloxidase ；melanin 中图分类号：Q554 文献标识码：A 文章编号：1002-6630(201