Structural Basis of Catalysis in the Bacterial Monoterpene Synthases Linalool Synthase and 1,8-Cineole Synthase.
Structural Basis of Catalysis in the Bacterial Monoterpene Synthases Linalool Synthase and 1,8-Cineole Synthase.
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DOI：
10.1021/acscatal.7b01924
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发表时间：
2017-09-01
期刊：
影响因子：
12.9
通讯作者：
Scrutton NS
中科院分区：
文献类型：
--
作者：
Karuppiah V;Ranaghan KE;Leferink NGH;Johannissen LO;Shanmugam M;Ní Cheallaigh A;Bennett NJ;Kearsey LJ;Takano E;Gardiner JM;van der Kamp MW;Hay S;Mulholland AJ;Leys D;Scrutton NS
萜类化合物形成了最大的和立体化学上最多样化的天然产物类别，并且通过用全细胞或纯化的酶进行生物催化以及通过代谢工程来生产这些具有相当大的兴趣。单萜是一类重要的萜类化合物，作为调味剂和香料在工业上是重要的。我们在这里报告的结构，最近发现的棒状链霉菌单萜脱氢酶芳樟醇合酶（bLinS）和1，8-桉叶素合酶（bCinS），我们表明，这些是活性生物催化剂的单萜生产使用生物催化和代谢工程平台。在代谢工程的单萜类产生菌E.在大肠杆菌菌株中，与相应的植物单萜合酶相比，使用bLinS导致芳樟醇产量高300倍。使用bCinS，1，8-桉叶素的纯度为96%，而植物物种的纯度为67%。bLinS和bCinS的结构，以及它们与氟化底物类似物的复合物，表明这些细菌单萜脱氢酶类似于先前表征的倍半萜脱氢酶。分子动力学模拟表明，这些单萜脱氢酶在反应周期中不会发生大规模的构象变化，使它们成为基于结构的蛋白质工程的有吸引力的目标，以扩大这些酶对替代单萜支架的催化范围。bLinS和bCinS结构的比较表明它们的活性位点如何将反应性碳阳离子中间体转向所需的无环芳樟醇（bLinS）或双环1，8-桉叶素（bCinS）产物。这里报道的工作提供了这类重要的单萜合酶的结构分析。这将指导细菌酶作为生产其他单萜和单萜类化合物的门户生物催化剂的开发。
Terpenoids form the largest and stereochemically most diverse class of natural products, and there is considerable interest in producing these by biocatalysis with whole cells or purified enzymes, and by metabolic engineering. The monoterpenes are an important class of terpenes and are industrially important as flavors and fragrances. We report here structures for the recently discovered Streptomyces clavuligerus monoterpene synthases linalool synthase (bLinS) and 1,8-cineole synthase (bCinS), and we show that these are active biocatalysts for monoterpene production using biocatalysis and metabolic engineering platforms. In metabolically engineered monoterpene-producing E. coli strains, use of bLinS leads to 300-fold higher linalool production compared with the corresponding plant monoterpene synthase. With bCinS, 1,8-cineole is produced with 96% purity compared to 67% from plant species. Structures of bLinS and bCinS, and their complexes with fluorinated substrate analogues, show that these bacterial monoterpene synthases are similar to previously characterized sesquiterpene synthases. Molecular dynamics simulations suggest that these monoterpene synthases do not undergo large-scale conformational changes during the reaction cycle, making them attractive targets for structured-based protein engineering to expand the catalytic scope of these enzymes toward alternative monoterpene scaffolds. Comparison of the bLinS and bCinS structures indicates how their active sites steer reactive carbocation intermediates to the desired acyclic linalool (bLinS) or bicyclic 1,8-cineole (bCinS) products. The work reported here provides the analysis of structures for this important class of monoterpene synthase. This should now guide exploitation of the bacterial enzymes as gateway biocatalysts for the production of other monoterpenes and monoterpenoids.
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影响因子：
5.5
作者：
Goetz, Andreas W.;Williamson, Mark J.;Xu, Dong;Poole, Duncan;Le Grand, Scott;Walker, Ross C.
通讯作者：
Walker, Ross C.
DOI：
10.1351/pac-con-09-09-37
发表时间：
2010
期刊：
Pure and applied chemistry. Chimie pure et appliquee
影响因子：
--
作者：
Aaron JA;Christianson DW
通讯作者：
Christianson DW
影响因子：
2.7
作者：
FUJISAKI, S;NISHINO, T;KATSUKI, H
通讯作者：
KATSUKI, H
影响因子：
11.9
作者：
Gao Y;Honzatko RB;Peters RJ
通讯作者：
Peters RJ
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH