Structure-based engineering of strictosidine synthase:: Auxiliary for alkaloid libraries
Structure-based engineering of strictosidine synthase:: Auxiliary for alkaloid libraries
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DOI：
10.1016/j.chembiol.2007.08.009
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发表时间：
2007-09-01
影响因子：
--
通讯作者：
Stoeckigt, Joachim
中科院分区：
文献类型：
--
作者：
Loris, Elke A.;Panjikar, Santosh;Stoeckigt, Joachim
高度底物特异性的异胡萝卜苷合成酶（EC 4.3.3.2）催化色胺和裂叶皂苷之间的生物Pictet-Spengler缩合，导致高等植物中约2000种单萜吲哚生物碱的合成。本文阐明了萝芙木strictosidine合酶（STR 1）与strictosidine复合物的晶体结构，从而使STR 1的活性中心发生合理的定点突变，并导致其底物接受度的调节。在这里，我们报告的STR 1的合理重新设计的代Val 208 Ala突变体，进一步描述底物接受和10-甲基和10-甲氧基strictosidines的酶催化合成的影响。基于从长春花细胞中提取的粗品异茉莉苷糖苷酶中加入异茉莉苷，提出了一种联合化学酶法产生大生物碱库的方法，以用于未来的药理学筛选。
The highly substrate-specific strictosidine synthase (EC 4.3.3.2) catalyzes the biological Pictet-Spengler condensation between tryptamine and secologanin, leading to the synthesis of about 2000 monoterpenoid indole alkaloids in higher plants. The crystal structure of Rauvolfia serpentina strictosidine synthase (STR1) in complex with strictosidine has been elucidated here, allowing the rational site-directed mutation of the active center of STR1 and resulting in modulation of its substrate acceptance. Here, we report on the rational redesign of STR1 by generation of a Val208Ala mutant, further describing the influence on substrate acceptance and the enzyme-catalyzed synthesis of 10-methyl- and 10-methoxystrictosidines. Based on the addition of strictosidine to a crude strictosidine glucosidase preparation from Catharanthus cells, a combined chemoenzy-matic approach to generating large alkaloid libraries for future pharmacological screenings is presented.