贝壳状革耳菌漆酶酶活测定方法分析

Methods of Assaying Laccase Activity by Panus Conchatus

酶活测定方法中所用的反应底物、反应条件以及酶活单位定义对漆酶酶活的测定结果有很大的影响。在对贝壳状革耳菌 (Panusconchatus)漆酶酶活测定方法的研究中发现 ,该酶与 2 ,2’ -连氮 -双 ( 3-乙基并噻- 6 -磺酸 ) (ABTS)反应的最适pH值为 3.0 ,而与紫丁香醛连氮、2 ,6-二甲氧基苯酚和邻甲联苯胺反应的最适pH值均为 4.0。上述底物与Panusconchatus漆酶反应的Km (米氏常数 )分别为 0 .0 1 1 6(mmol·L) - 1 ,1 4.1 0 95 (mmol·L) - 1 ,0 .1 0 1 1 (mmol·L) - 1 ,0 .1 41 5 (mmol·L) - 1 。该酶与ABTS、2 ,6-二甲氧基苯酚和邻甲联苯胺反应均表现为零级反应 ,而与紫丁香醛连氮反应时只在一定范围的酶浓度以及反应时间内才表现为零级反应。

The data of laccase activity was greatly different according to the definition of laccase activity unit pH and substrates in reaction. The optimum pH for Panus conchatus laccase reacting with 2,6-dimethoxyphenol, o-tolidine, syringaldazine and ABTS were 4.0, 4.0, 4.0, 3.0 respectively, and the Km were 0.0116(mmol·L) -1 , 0.1011(mmol·L) -1 , 0.1415(mmol·L) -1 , 14.1095(mmol·L) -1 separately. It was also found that syringaldzine was unfit for Panus conchatus laccase activity assay.