Science paper Genome-based optimization of psilocybin and N,N-dimethyltryptamine biosynthetic pathways in E. coli using CRISPR-associated transposases

[d*l*b]

[Elsevier] Genome-based optimization of psilocybin and N,N-dimethyltryptamine biosynthetic pathways in E. coli using CRISPR-associated transposases

Abstract

Stable, high-level biosynthesis of complex natural products requires precise control of heterologous pathway expression, yet transcriptional architectures optimized on plasmids often fail when transferred to the chromosome. Here, we present ePathIntegrate, a genome-centric pathway engineering strategy that leverages CRISPR-associated transposases (CASTs) to integrate and rebalance multigene metabolic pathways in Escherichia coli. Direct genomic transfer of plasmid-optimized psilocybin and N,N-dimethyltryptamine (DMT) pathways resulted in a loss of productivity, driven by context-dependent promoter behavior. To address this, we developed and characterized a library of mutant T7 promoters that restore mid-range transcriptional control on the genome. Applying ePathIntegrate enabled re-optimization of both pathways, yielding genome-encoded strains that achieve 1.88 g/L psilocybin and 1.62 g/L DMT in fed-batch bioreactors. Whole-genome sequencing of CAST-mediated strains further revealed (i) precise on-target integration, (ii) some off-target pathway integrations, and (iii) small mutations in a subset of strains, highlighting both the power and limitations of CAST-mediated strain engineering.

https://www.sciencedirect.com/science/article/pii/S1096717626000856

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[Nydex]

That's really cool actually, holy crap! 1.88 g/L psilocybin and 1.62 g/L DMT are insane yields, considering how little a liter is. I can clearly see a future where getting a hold of these medicines is easier than ever, and honestly this makes me feel a bit more optimistic about humanity's fate. Sure, these medicines don't work equally well on everyone, but it's sure as hell better than drinking alcohol and sofa rotting.

I'm not smart enough to understand exactly how they achieved what they did, but man this kind of tech is like magic to me, so exciting! Thanks for sharing!