Cytochrome P450-modified bacterial terpenoids remain in a vast chemical space to be explored. In the present study, we conducted global genome mining of 223,829 bacterial genomes and identified 2892 bacterial terpenoid biosynthetic gene clusters (BGCs) with cytochrome P450 genes. Among these, we selected 562 with multiple P450 enzymes, which were further clustered as 355 gene cluster families by sequence similarity analysis. We then chose lev, a BGC from Streptomyces levis MCCC1A01616, for heterologous expression and discovered four new α-amorphene-type sesquiterpenoids, levinoids A–D (1–4). The structures and absolute configurations of these four new compounds were determined by employing extensive NMR analysis, NMR chemical shift calculations with DP4+, and ECD calculations. Furthermore, levinoid C (3) exhibited a moderate level of neuroprotective activity (EC₅₀ = 21 μM) in the glutamate-induced excitotoxicity cell model. Our findings highlight the untapped chemical diversity of P450-modified bacterial terpenoids, opening new avenues for further exploration and discovery.

中文翻译：

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通过全球基因组挖掘发现 P450 修饰的倍半萜类化合物 Levinoids A–D

细胞色素 P450 修饰的细菌萜类化合物仍有广阔的化学空间有待探索。在本研究中，我们对223,829个细菌基因组进行了全球基因组挖掘，并鉴定了2892个具有细胞色素P450基因的细菌萜类生物合成基因簇（BGC）。其中，我们筛选出562个具有多种P450酶的基因，通过序列相似性分析将其进一步聚类为355个基因簇家族。然后，我们选择lev （来自Streptomyces levis MCCC1A01616 的BGC）进行异源表达，并发现了四种新的 α-amorphene 型倍半萜类化合物 levinoids A–D ( 1 – 4 )。这四种新化合物的结构和绝对构型通过广泛的 NMR 分析、DP4+ 的 NMR 化学位移计算和 ECD 计算来确定。此外，Levinoid C ( 3 )在谷氨酸诱导的兴奋性毒性细胞模型中表现出中等水平的神经保护活性（EC ₅₀ = 21 μM）。我们的研究结果强调了 P450 修饰的细菌萜类化合物尚未开发的化学多样性，为进一步探索和发现开辟了新途径。