In vivo production of psilocybin in E. coli

Alexandra M Adams; Nicholas A Kaplan; Zhangyue Wei; John D Brinton; Chantal S Monnier; Alexis L Enacopol; Theresa A Ramelot; J Andrew Jones

doi:10.1016/j.ymben.2019.09.009

In vivo production of psilocybin in E. coli

Metab Eng. 2019 Dec:56:111-119. doi: 10.1016/j.ymben.2019.09.009. Epub 2019 Sep 21.

Authors

Alexandra M Adams¹, Nicholas A Kaplan², Zhangyue Wei³, John D Brinton⁴, Chantal S Monnier⁵, Alexis L Enacopol⁶, Theresa A Ramelot⁷, J Andrew Jones⁸

Affiliations

¹ Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: adamsam8@miamioh.edu.
² Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: kaplanna@miamioh.edu.
³ Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: weiz2@miamioh.edu.
⁴ Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: brintojd@miamioh.edu.
⁵ Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: monniecs@miamioh.edu.
⁶ Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: enacopal@miamioh.edu.
⁷ Miami University, Department of Chemistry and Biochemistry, Oxford, OH, 45056, USA. Electronic address: t.ramelot@miamioh.edu.
⁸ Miami University, Department of Chemical, Paper, And Biomedical Engineering, Oxford, OH, 45056, USA. Electronic address: jonesj28@miamioh.edu.

PMID: 31550507
DOI: 10.1016/j.ymben.2019.09.009

Abstract

Psilocybin, the prodrug of the psychoactive molecule psilocin, has demonstrated promising results in clinical trials for the treatment of addiction, depression, and post-traumatic stress disorder. The development of a psilocybin production platform in a highly engineerable microbe could lead to rapid advances towards the bioproduction of psilocybin for use in ongoing clinical trials. Here, we present the development of a modular biosynthetic production platform in the model microbe, Escherichia coli. Efforts to optimize and improve pathway performance using multiple genetic optimization techniques were evaluated, resulting in a 32-fold improvement in psilocybin titer. Further enhancements to this genetically superior strain were achieved through fermentation optimization, ultimately resulting in a fed-batch fermentation study, with a production titer of 1.16 g/L of psilocybin. This is the highest psilocybin titer achieved to date from a recombinant organism and a significant step towards demonstrating the feasibility of industrial production of biologically-derived psilocybin.

Keywords: Baeocystin; Escherichia coli; Hallucinogenic; Metabolic engineering; Norbaeocystin; Pathway balancing; Promoter library; Psilocybin; Psychedelic; Recombinant production.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Batch Cell Culture Techniques*
Escherichia coli* / genetics
Escherichia coli* / growth & development
Metabolic Engineering*
Psilocybin* / biosynthesis
Psilocybin* / genetics

Substances

Psilocybin