Abstract：
Genome mining of the Actinomycete Crossiella cryophila facilitated the discovery of a minimal terpenoid biosynthetic gene cluster of cry consisting of a class I terpene cyclase CryA and a CYP450 monooxygenase CryB. Heterologous expression of cry allowed the isolation and characterization of two new sesquiterpenoids, ent-viridiflorol (1) and cryophilain (2). Notably, cryophilain (2) possesses a 5/7/3-fused tricyclic skeleton bearing a distinctive bridgehead hydroxy group. The combined in vivo and in vitro experiments revealed that CryA, the first ent-viridiflorol terpene cyclase, catalyzes farnesyl diphosphate to form the 5/7/3 sesquiterpene core scaffold and P450 CryB serves as a tailoring enzyme responsible for installing a hydroxy group at the bridgehead carbon.
摘要：
放线菌Crossiellacryila的基因组挖掘促进了cry的最小萜类生物合成基因簇的发现，该簇由I类萜烯环化酶CryA和CYP450单加氧酶CryB组成。cry的异源表达允许分离和表征两种新的倍半萜类化合物，ent-viridiflorol（1）和冷冻（2）。值得注意的是,冷冻法（2）具有5/7/3稠合的三环骨架，带有独特的桥头羟基。体内和体外实验相结合，表明CryA，第一个ENT-viridflorol萜烯环化酶，催化法尼基二磷酸以形成5/7/3倍半萜烯核心支架，P450CryB充当剪裁酶，负责在桥头碳上安装羟基。
