Srivastava Prabhakar L, Johns Sam T, Walters Rebecca, Miller David J, Van der Kamp Marc W, Allemann Rudolf K
School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff, CF10 3AT, United Kingdom.
School of Biochemistry, University of Bristol, University Walk, Bristol, BS8 1TD, United Kingdom.
ACS Catal. 2023 Oct 20;13(21):14199-14204. doi: 10.1021/acscatal.3c03920. eCollection 2023 Nov 3.
Terpene synthases (TS) catalyze complex reactions to produce a diverse array of terpene skeletons from linear isoprenyl diphosphates. Patchoulol synthase (PTS) from converts farnesyl diphosphate into patchoulol. Using simulation-guided engineering, we obtained PTS variants that eliminate water capture. Further, we demonstrate that modifying the structurally conserved Hα-1 loop also reduces hydroxylation in PTS, as well as in germacradiene-11-ol synthase (Gd11olS), leading to cyclic neutral intermediates as products, including α-bulnesene (PTS) and isolepidozene (Gd11olS). Hα-1 loop modification could be a general strategy for engineering sesquiterpene synthases to produce complex cyclic hydrocarbons without the need for structure determination or modeling.
萜烯合酶（TS）催化复杂反应，从线性异戊烯基二磷酸生成各种各样的萜烯骨架。来自[具体来源未提及]的广藿香醇合酶（PTS）将法尼基二磷酸转化为广藿香醇。通过模拟引导工程，我们获得了消除水捕获的PTS变体。此外，我们证明修饰结构保守的Hα-1环也能减少PTS以及牻牛儿二烯-11-醇合酶（Gd11olS）中的羟基化，从而产生环状中性中间体作为产物，包括α-布藜烯（PTS）和异lepidozene（Gd11olS）。Hα-1环修饰可能是一种通用策略，用于工程化倍半萜烯合酶以生产复杂的环状烃，而无需进行结构测定或建模。
