The lid is a structural and functional determinant of lipase activity and selectivity
The lid is a structural and functional determinant of lipase activity and selectivity
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DOI：
10.1016/j.molcatb.2006.01.018
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发表时间：
2006-05-02
影响因子：
--
通讯作者：
Eggert, T
中科院分区：
文献类型：
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作者：
Secundo, F;Carrea, G;Eggert, T
在几种脂肪酶中，酶活性部位的进入受称为盖的移动的结构的位置调节。本文综述了这一区域在调节脂肪酶功能中的作用，分析了3种不同的重组脂肪酶：Candida rugosa lipase isoform 1（CRL 1），Pseudomonasfragi lipase（PFL）和Bacillussubtilis lipase A（BSLA）的结果。通过用另一种C.结果表明，在有机溶剂中，Rugosa脂肪酶异构体（CRL 1LID 3）的活性和对映体选择性均受到影响。PFL蛋白的变体，其中三个极性盖残基被替换为在同源脂肪酶中严格保守的氨基酸显示改变的链长偏好曲线和增加的热稳定性。另一方面，插入盖结构从结构同源酶BSLA，脂肪酶，天然不具有这样的盖结构，引起酶活性的降低和底物特异性的改变。这些结果有力地支持了这样的概念，即盖子不仅在调节脂肪酶的活性，而且在调节脂肪酶的特异性、对映选择性和稳定性方面起着重要作用。(c)2006 Elsevier B.V.保留所有权利。
In several lipases access to the enzyme active site is regulated by the position of a mobile structure named the lid. The role of this region in modulating lipase function is reviewed in this paper analysing the results obtained with three different recombinant lipases modified in the lid sequence: Candida rugosa lipase isoform 1 (CRL1), Pseudomonas fragi lipase (PFL) and Bacillus subtilis lipase A (BSLA). A CRL chimera enzyme obtained by replacing its lid with that of another C. rugosa lipase isoform (CRL1LID3) was found to be affected in both activity and enantioselectivity in organic solvent. Variants of the PFL protein in which three polar lid residues were replaced with amino acids strictly conserved in homologous lipases displayed altered chain length preference profile and increased thermostability. On the other hand, insertion of lid structures from structurally homologous enzymes into BSLA, a lipase that naturally does not possess such a lid structure, caused a reduction in the enzyme activity and an altered substrate specificity. These results strongly support the concept that the lid plays an important role in modulating not only activity but also specifity, enantioselectivity and stability of lipase enzymes. (c) 2006 Elsevier B.V. All rights reserved.