Yeast-engineered biocatalysis enables stepwise production of MDMA and analogs

By integrating engineered enzymes into a stepwise production platform, the researchers demonstrated a proof-of-concept route that replaces traditional chemical synthesis with biotechnology.

MDMA became widely known in the 1970s for its psychoactive and mood-enhancing properties. Initially popularized as a psychotherapy aid, it later spread as a recreational drug associated with nightlife and electronic music scenes. Regulatory crackdowns in the mid-1980s classified MDMA as a Schedule I substance, limiting its legal use and hindering research. In recent years, however, scientific and clinical interest in MDMA has resurged. Clinical trials have shown that MDMA-assisted therapy can be highly effective for patients with severe or treatment-resistant post-traumatic stress disorder (PTSD). A new drug application for MDMA-assisted therapy has already been filed with the U.S. Food and Drug Administration and granted priority review. Despite this momentum, MDMA production still relies on controlled chemical precursors such as safrole and piperonal, presenting regulatory and environmental challenges. Biotechnology offers a promising alternative, as advances in synthetic biology have enabled microbial production of compounds like psilocybin and mescaline. Yet, MDMA’s semi-synthetic nature has so far required innovative new strategies.

A study (DOI:10.1016/j.bidere.2025.100011) published in BioDesign Research on 21 March 2025 by Peter J. Facchini’s team, Enveric Biosciences Inc, could lay the foundation for safer, more sustainable production of psychoactive compounds being investigated for their therapeutic potential, particularly in treating PTSD.

Using a stepwise hybrid bioconversion–biocatalysis strategy in yeast, the researchers first screened multiple pyruvate decarboxylases (PDCs) and rationally engineered variants to carboligate pyruvate with aromatic aldehydes, then evaluated stereoselective ω-transaminases to aminate the resulting phenylacetylcarbinol (PAC) ketones, next applied an N-methyltransferase to generate N-methylated phenylpropylamines, and finally used an analytical confirmation of product formation after a chemical reduction step. Corresponding to this workflow, the PDC screen identified Candida tropicalis PDC as the most robust catalyst, with an I479A substitution further boosting acceptance of ring-substituted substrates and PAC formation; modest medium optimization increased PAC titers, and the platform converted piperonal and a broad panel of benzaldehyde analogs, with 15/23 substrates accepted and isolated PAC derivatives typically reaching ~20–70% conversion (piperonal ≈47.5%; chlorination at the 6-position reduced yield). In the amination phase, three engineered ω-transaminases were compared; ATA-117-Rd11 consistently outperformed VfTA-M7 and BPTA-M4 across PAC variants, enabling isolation of multiple aminated intermediates in vitro (though in-cell execution was limited by unfavorable thermodynamics and insufficient flux). For N-methylation, human phenylethanolamine N-methyltransferase accepted the aminated products—including the MDMA intermediate and a chlorinated analog—demonstrating step compatibility but revealing a second rate limitation due to low catalytic efficiency on these non-native substrates. To close the sequence, reduction of the N-methylated intermediates yielded MDMA and 6-chloro-MDMA detectable by targeted mass-spectrometric transitions, validating the end-to-end feasibility of the route. Overall, methodical enzyme selection/engineering, substrate-scope expansion, and orthogonal in vitro steps produced milligram-scale, structurally verified intermediates and final products, while pinpointing two principal bottlenecks for future optimization: (i) driving ω-transamination forward in vivo and (ii) enhancing N-methyltransferase activity and breadth for phenylpropylamine scaffolds.

The successful demonstration of MDMA bioproduction carries significant implications for both medicine and industry. If optimized, microbial fermentation platforms could provide a safer and more sustainable supply chain for clinical-grade MDMA, reducing reliance on environmentally damaging or tightly controlled chemical precursors. Beyond MDMA, the approach may be extended to generate novel derivatives with improved therapeutic profiles, such as analogs with reduced side effects or enhanced efficacy. Moreover, this proof-of-concept strengthens the broader role of synthetic biology in reimagining pharmaceutical production.

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References

DOI

10.1016/j.bidere.2025.100011

Original Source URL

https://doi.org/10.1016/j.bidere.2025.100011

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