Abstract：
N,N-dimethyltryptamine (DMT), 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) and 5-hydroxy-N,N-dimethyltryptamine (bufotenine) are psychedelic tryptamines found naturally in both plants and animals and have shown clinical potential to help treat mental disorders, such as anxiety and depression. Advances in both metabolic and genetic engineering make it possible to engineer microbes as cell factories to produce DMT and its aforementioned derivatives to meet demand for ongoing clinical study. Here, we present the development of a biosynthetic production pathway for DMT, 5-MeO-DMT, and bufotenine in the model microbe Escherichia coli. Through the application of genetic optimization techniques and process optimization in benchtop fermenters, the in vivo production of DMT in E. coli was observed. DMT production with tryptophan supplementation reached maximum titers of 74.7 ± 10.5 mg/L under fed batch conditions in a 2-L bioreactor. Additionally, we show the first reported case of de novo production of DMT (from glucose) in E. coli at a maximum titer of 14.0 mg/L and report the first example of microbial 5-MeO-DMT and bufotenine production in vivo. This work provides a starting point for further genetic and fermentation optimization studies with the goal to increase methylated tryptamine production metrics to industrially competitive levels.
摘要：
N,N:二甲基色胺(DMT),5-甲氧基-N,N-二甲基色胺（5-MeO-DMT）和5-羟基-N，N-二甲基色胺（bufotenine）是在植物和动物中天然发现的迷幻色胺，并已显示出有助于治疗精神障碍的临床潜力，如焦虑和抑郁。代谢和基因工程的进步使得有可能将微生物改造为细胞工厂以生产DMT及其上述衍生物，以满足正在进行的临床研究的需求。这里,我们提出了DMT生物合成生产途径的发展，5-MeO-DMT,和模式微生物大肠杆菌中的bufotenine。经由过程遗传优化技巧和工艺优化在台式发酵罐中的应用，观察到在大肠杆菌中DMT的体内产生。在2-L生物反应器中的分批补料条件下，添加色氨酸的DMT生产达到74.7±10.5mg/L的最大滴度。此外,我们显示了在大肠杆菌中以14.0mg/L的最大滴度从头产生DMT（来自葡萄糖）的第一个报道案例，并报道了微生物5-MeO-DMT和bufotenine在体内产生的第一个例子。这项工作为进一步的遗传和发酵优化研究提供了起点，目的是将甲基化色胺的生产指标提高到工业竞争水平。
