Biosynthesis of sodorifen with a unique C₁₆-bicyclo[3.2.1]octene framework requires an S-adenosyl methionine-dependent methyltransferase SodC and terpene cyclase SodD. While bioinformatic analyses reveal a wide distribution of the sodCD genes organization in bacteria, their functional diversity remains largely unknown. Herein, two sodorifen-type gene clusters, pcch and pcau, from Pseudomonas sp. are heterologously expressed in Escherichia coli, leading to the discovery of two C₁₆ terpenoids. Enzymatic synthesis of these compounds is achieved using the two (SodCD-like) pathway-specific enzymes. Enzyme assays using different combinations of methyltransferases and terpene synthases across the pcch, pcau, and sod pathways reveal a unifying biosynthetic mechanism: all three SodC-like enzymes methylate farnesyl pyrophosphate (FPP) with subsequent cyclization to a common intermediate, pre-sodorifen pyrophosphate. Structural diversification of this joint precursor solely occurs by the subsequently acting individual terpene synthases. Our findings expand basic biosynthetic understanding and structural diversity of unusual C₁₆-terpenoids.

中文翻译：

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来自假单胞菌的非经典 C16-萜类化合物的发现和生物合成


具有独特的C ₁₆ -双环[3.2.1]辛烯框架的sodorifen的生物合成需要S-腺苷甲硫氨酸依赖性甲基转移酶SodC和萜烯环化酶SodD。虽然生物信息学分析揭示了sodCD基因组织在细菌中的广泛分布，但它们的功能多样性仍然很大程度上未知。在此，来自假单胞菌属的两个 sodorifen 型基因簇pcch和pcau 。在大肠杆菌中异源表达，从而发现了两种 C ₁₆萜类化合物。这些化合物的酶促合成是使用两种（SodCD 样）途径特异性酶来实现的。使用跨pcch 、 pcau和sod途径的甲基转移酶和萜烯合酶的不同组合进行的酶测定揭示了统一的生物合成机制：所有三种 SodC 样酶均甲基化法尼基焦磷酸 (FPP)，随后环化为常见的中间体，前 sodorifen 焦磷酸。该联合前体的结构多样化仅由随后起作用的各个萜烯合酶发生。我们的研究结果扩展了对不寻常的 C ₁₆萜类化合物的基本生物合成理解和结构多样性。