胱氨酸与烟草多酚氧化酶相互作用的研究

Study on the Interaction of Cystine and Polyphenol Oxidase from Nicotian Tobaccum

研究了胱氨酸与烟草多酚氧化酶的相互作用。结果表明 ,胱氨酸对烟草多酚氧化酶有抑制作用 ,其机制可能是胱氨酸中的硫醚与酶活性中心铜离子配位 ,但反应物或产物均抑制这种络合 ;胱氨酸还可以与产物结合生成无色复合物 ;当胱氨酸与多酚氧化酶的分子比达到 16 0 0 0∶1时 ,就完全抑制了酶活性 ;对于荧光光谱 ,当分子比为 75∶1时 ,多酚氧化酶的荧光就不再发生变化 ;烟草多酚氧化酶中的色氨酸残基位于比较疏水的环境。

The interaction of cystine and polyphenol oxidase(PPO) from nicotian tobaccum has been studied. The results show that cystine has an inhibitory effect on the enzymatic activity, the mechanism of which might be that the thiolether in cystine coordinates with Cu2+ in the active site of PPO, and the inactivation constant value of PPO by cystine is 0.633 min(-1), while the substrates or the products inhibit their combination. Cystine can also be combined with the products of enzymatic reaction to form a colourless compound. Cystine can inhibit the enzymatic activity completely when the molar ratio of cystine to PPO approaches 16 000:1. The effect of cystine on the fluorescence changes with the molar ratio of cystine to the PPO. However, when the molar ratio gets to 75: 1, cystine has no longer the effect on either the fluorescence spectra or the synchronous spectra. Microenvironment of trp residue in PPO is more hydrophobic than that of free trp in water.