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Intein-mediated temperature control for complete biosynthesis of sanguinarine and its halogenated derivatives in yeast.

Yuanwei GouDongfang LiMinghui ZhaoMengxin LiJiaojiao ZhangYilian ZhouFeng XiaoGaofei LiuHaote DingChenfan SunCuifang YeChang DongJucan GaoDi GaoZehua BaoLei HuangZhinan XuJiazhang Lian
Published in: Nature communications (2024)
While sanguinarine has gained recognition for antimicrobial and antineoplastic activities, its complex conjugated structure and low abundance in plants impede broad applications. Here, we demonstrate the complete biosynthesis of sanguinarine and halogenated derivatives using highly engineered yeast strains. To overcome sanguinarine cytotoxicity, we establish a splicing intein-mediated temperature-responsive gene expression system (SIMTeGES), a simple strategy that decouples cell growth from product synthesis without sacrificing protein activity. To debottleneck sanguinarine biosynthesis, we identify two reticuline oxidases and facilitated functional expression of flavoproteins and cytochrome P450 enzymes via protein molecular engineering. After comprehensive metabolic engineering, we report the production of sanguinarine at a titer of 448.64 mg L -1 . Additionally, our engineered strain enables the biosynthesis of fluorinated sanguinarine, showcasing the biotransformation of halogenated derivatives through more than 15 biocatalytic steps. This work serves as a blueprint for utilizing yeast as a scalable platform for biomanufacturing diverse benzylisoquinoline alkaloids and derivatives.
Keyphrases
  • cell wall
  • gene expression
  • structure activity relationship
  • saccharomyces cerevisiae
  • escherichia coli
  • staphylococcus aureus
  • poor prognosis
  • amino acid
  • wastewater treatment