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McKinney_Thesis_2023.pdf (3.55 MB)

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Screening of Norbaeocystin Methyltransferase Variants Enables Enhanced Psilocybin and Baeocystin Production in E. coli

McKinney, Madeline Gray
http://orcid.org/0009-0004-9109-2154
http://rave.ohiolink.edu/etdc/view?acc_num=miami1682002450590186

Abstract Details

2023, Master of Science, Miami University, Chemical, Paper and Biomedical Engineering.
Clinical evaluation of psilocybin, a hallucinogen found in “magic mushrooms,” as treatment for severe depression and addiction is ongoing. Our enzymatic pathway to production of psilocybin involves norbaeocystin methyltransferase (PsiM), responsible for the iterative methylation of norbaeocystin, resulting in the production of psilocybin via monomethylated intermediate, baeocystin. Genetic variation between magic mushroom species have motivated this study to evaluate multiple PsiM variants for their ability to produce target compounds. Here, we examine the effect of genetic variability of the PsiM gene and attempt to create a biosynthetic production platform for producing higher titers of psilocybin than previously reported and strains capable of baeocystin-specific production. Screening of genes from Psilocybe cubensis, Psilocybe cyanescens, Panaeolus cyanescens, Gymnopilus dilepis and Gymnopilus junonius yielded production strains capable of exceeding 490 mg/L of psilocybin before optimization. Following fed-batch bioreactor scale-up, production of over 1.46 ± 0.13 g/L psilocybin was observed, surpassing the current benchmark titer of 1.16 g/L. This construct employs a constitutive promoter, enhancing the economic viability by removing the need for inducing chemicals. This work further enhances the viability of industrial batch production of psilocybin using fermentation and provides a basis for development of selective precursor production strains for combined medicinal use.
J. Andrew Jones (Advisor)
Jason Berberich (Committee Member)
Kevin Yehl (Committee Member)
Justin Saul (Committee Member)
40 p.
Biomedical Engineering; Chemical Engineering
Norbaeocystin Methyltransferase; Psilocybin; Baeocystin; PsiM; Metabolic Engineering; Synthetic Biology;

Recommended Citations

Citations

  • McKinney, M. G. (2023). Screening of Norbaeocystin Methyltransferase Variants Enables Enhanced Psilocybin and Baeocystin Production in E. coli [Master's thesis, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1682002450590186

    APA Style (7th edition)

  • McKinney, Madeline. Screening of Norbaeocystin Methyltransferase Variants Enables Enhanced Psilocybin and Baeocystin Production in E. coli. 2023. Miami University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1682002450590186.

    MLA Style (8th edition)

  • McKinney, Madeline. "Screening of Norbaeocystin Methyltransferase Variants Enables Enhanced Psilocybin and Baeocystin Production in E. coli." Master's thesis, Miami University, 2023. http://rave.ohiolink.edu/etdc/view?acc_num=miami1682002450590186

    Chicago Manual of Style (17th edition)

miami1682002450590186
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© 2023, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.
Release 3.2.12