Psilocybin is a tryptamine-derived psychoactive alkaloid found mainly in the fungal genus Psilocybe, among others, and is the active ingredient in so-called “magic mushrooms”. Although its notoriety originates from its psychotropic properties and popular use as a recreational drug, clinical trials have recently recognized psilocybin as a promising candidate for the treatment of various psychological and neurological afflictions. In this work, we demonstrate the de novo biosynthetic production of psilocybin and related tryptamine derivatives in Saccharomyces cerevisiae by expression of a heterologous biosynthesis pathway sourced from Psilocybe cubensis. Additionally, we achieve improved product titers by supplementing the pathway with a novel cytochrome P450 reductase from P. cubensis. Further rational engineering resulted in a final production strain producing 627 ± 140 mg/L of psilocybin and 580 ± 276 mg/L of the dephosphorylated degradation product psilocin in triplicate controlled fed-batch fermentations in minimal synthetic media. Pathway intermediates baeocystin, nor norbaeocystin as well the dephosphorylated baeocystin degradation product norpsilocin were also detected in strains engineered for psilocybin production. We also demonstrate the biosynthetic production of natural tryptamine derivative aeruginascin as well as the production of a new-to-nature tryptamine derivative N-acetyl-4-hydroxytryptamine. These results lay the foundation for the biotechnological production of psilocybin in a controlled environment for pharmaceutical applications, and provide a starting point for the biosynthetic production of other tryptamine derivatives of therapeutic relevance.

赛洛西宾是一种来源于色胺的精神活性生物碱，主要存在于真菌属赛洛西比中，并且是所谓“魔菇”中的活性成分。尽管它的臭名昭著源于它的精神特性和作为娱乐性药物的广泛应用，但临床试验最近已将psilocybin视为治疗各种心理和神经疾病的有前途的候选者。在这项工作中，我们通过表达源自立方假单胞菌的异源生物合成途径来证明啤酒酵母中从头生物合成psilocybin和相关色胺的衍生物。此外，我们还通过向该途径添加了来自立方体假单胞菌的新型细胞色素P450还原酶来提高了产品效价。进一步的合理工程设计导致最终生产菌株在最少的合成培养基中进行了三次重复的补料分批发酵，产生627±140 mg / L psilocybin和580±276 mg / L脱磷酸降解产物psilocin。在为产生psilocybin产生而设计的菌株中，也检测到了途径中间体baeocystin，norbaeocystin以及去磷酸化baeocystin降解产物norpsilocin。我们还演示了自然色胺素衍生物铜绿素的生物合成生产以及新的自然色胺素衍生物N-的生产。乙酰-4-羟基色胺。这些结果为在药物应用的受控环境中生物倍半霉素的生物技术生产奠定了基础，并为生物合成生产具有治疗意义的其他色胺衍生物提供了起点。