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<p>Rotenoids are structurally complex isoflavones primarily produced by leguminous plants. These natural products are widely used as insecticides and piscicides in agriculture and are also of interest for their antiviral and anticancer properties. Despite their extensive use, the biosynthetic pathway of rotenoids has remained elusive. Here we report the complete biosynthetic pathway of rotenoids in the Fabaceae plants <i>Amorpha fruticosa</i> and <i>Tephrosia vogelii</i>, revealing that the characteristic <i>cis</i>-fused B/C ring core structure is formed through a unique redox-neutral cyclization catalysed by an Fe(II)/α-ketoglutarate-dependent dioxygenase. The diverse E-ring structures are constructed by two distinct families of oxidases, cytochrome P450 enzymes in <i>A. fruticosa</i> and flavin adenine dinucleotide-dependent oxidase in <i>T. vogelii</i>. Furthermore, we leveraged the newly identified genes to engineer de novo rotenoid biosynthesis in the heterologous host <i>Nicotiana benthamiana</i>. This study provides fundamental insights into the biosynthesis of rotenoids and paves the way for the development of commercially valuable rotenoid-based insecticides through synthetic biology.</p><p></p>

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Discovery and engineering of the biosynthesis of rotenoids

  • 期刊论文

  • 发表日期：

    2025年10月21日

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  • Wenying Cao,
  • Jie Yang,
  • Benke Hong

摘要

Rotenoids are structurally complex isoflavones primarily produced by leguminous plants. These natural products are widely used as insecticides and piscicides in agriculture and are also of interest for their antiviral and anticancer properties. Despite their extensive use, the biosynthetic pathway of rotenoids has remained elusive. Here we report the complete biosynthetic pathway of rotenoids in the Fabaceae plants Amorpha fruticosa and Tephrosia vogelii, revealing that the characteristic cis-fused B/C ring core structure is formed through a unique redox-neutral cyclization catalysed by an Fe(II)/α-ketoglutarate-dependent dioxygenase. The diverse E-ring structures are constructed by two distinct families of oxidases, cytochrome P450 enzymes in A. fruticosa and flavin adenine dinucleotide-dependent oxidase in T. vogelii. Furthermore, we leveraged the newly identified genes to engineer de novo rotenoid biosynthesis in the heterologous host Nicotiana benthamiana. This study provides fundamental insights into the biosynthesis of rotenoids and paves the way for the development of commercially valuable rotenoid-based insecticides through synthetic biology.