Characterization of monophenolase activity of polyphenol oxidase from iceberg lettuce

被引:14
作者
Chazarra, S [1 ]
García-Carmona, F [1 ]
Cabanes, J [1 ]
机构
[1] Univ Murcia, Dept Bioquim & Biol Mol A, Unidad Docente Biol, Edificio Vet, E-30071 Murcia, Spain
关键词
plant polyphenol oxidase; lettuce; Lactuca sativa; monophenolase activity;
D O I
10.1021/jf980859o
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
Polyphenoloxidase (EC 1.14.18.1), a thylakoid membrane-bound enzyme, was isolated by sonication of osmotically shocked chloroplasts from iceberg lettuce (Lactuca sativa). The enzyme showed monophenolase activity when assayed on (p-hydroxyphenyl)propionic acid with 3-methyl-2-benzothiazolinone hydrazone in a reliable continuous spectrophotometric method, with high sensitivity, accuracy, and precision. The monophenolase activity showed a lag period before the steady-state rate (V-ss) was reached. Both kinetic parameters, the lag period and the steady-state rate, depended on the pH, the enzyme and substrate concentrations, and the presence of catalytic amounts of o-diphenol. This activity shows inhibition by high substrate concentration. The experimental results correspond with the mechanism previously described for PPO from other sources. Kinetic constants K-m, V-max, and K-i were determined.
引用
收藏
页码:1422 / 1426
页数:5
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