Lipase from Thermomyces lanuginosus (TLL) has been covalently immobilized on heterofunctional octyl-vinyl agarose. That way, the covalently immobilized enzymes will have identical orientation. Then, it has blocked using hexyl amine (HEX), ethylenediamine (EDA), Gly and Asp. The initial activity/stability of the different biocatalysts was very different, being the most stable the biocatalyst blocked with Gly. These biocatalysts had been utilized to analyze if the enzyme activity could decrease differently along thermal inactivation courses depending on the utilized substrate (that is, if the enzyme specificity was altered during its inactivation using 4 different substrates to determine the activity), and if this can be altered by the nature of the blocking agent and the inactivation conditions (we use pH 5, 7 and 9). Results show great changes in the enzyme specificity during inactivation (e.g., activity versus triacetin was much more quickly lost than versus the other substrates), and how this was modulated by the immobilization protocol and inactivation conditions. The difference in the changes induced by immobilization and inactivation were confirmed by fluorescence studies. That is, the functional and structural analysis of partially inactivated immobilized enzyme showed that their inactivation pathway is strongly depended on the support features and inactivation conditions.

来自绵毛嗜热丝孢菌的脂肪酶(TLL) 已共价固定在异功能的辛基-乙烯基琼脂糖上。这样，共价固定的酶将具有相同的方向。然后，它使用己胺 (HEX)、乙二胺 (EDA)、Gly 和 Asp 进行封闭。不同生物催化剂的初始活性/稳定性非常不同，用 Gly 封闭的生物催化剂最稳定。这些生物催化剂已被用于分析酶活性是否会随着热灭活过程的不同而降低，这取决于所用的底物（即，在使用 4 种不同的底物确定活性的灭活过程中，酶的特异性是否发生变化），以及这是否可以会因封闭剂的性质和灭活条件而改变（我们使用 pH 值 5、7 和 9）。结果显示灭活过程中酶的特异性发生了巨大变化（例如，与三醋精相比，活性比与其他底物相比丧失得更快），以及这是如何通过固定化方案和灭活条件进行调节的。荧光研究证实了固定化和失活引起的变化的差异。也就是说，部分灭活的固定化酶的功能和结构分析表明，它们的灭活途径在很大程度上取决于支持特征和灭活条件。荧光研究证实了固定化和失活引起的变化的差异。也就是说，部分灭活的固定化酶的功能和结构分析表明，它们的灭活途径在很大程度上取决于支持特征和灭活条件。荧光研究证实了固定化和失活引起的变化的差异。也就是说，部分灭活的固定化酶的功能和结构分析表明，它们的灭活途径在很大程度上取决于支持特征和灭活条件。