This project is aimed at introducing the heterologous biosynthetic pathway for psilocybin production in an existing GSM of A. niger, and thereafter use computational methods to identify which changes to the model could optimize production. Cofactor swap optimization revealed swapping cofactor specificity of two reactions in the model from NADPH to NADH could potentially improve theoretical yield of psilocybin. Investigation of gene regulation targets revealed several reactions that could be either upregulated or downregulated to improve psilocybin production flux. Two of the enzymes targeted for upregulation are in the shikimate pathway, which leads to the psilocybin precursor tryptophan, and were also shown to improve psilocybin production in previous studies. Several new gene prediction targets for upregulation, including methylation reactions, were found that could lead to promising results in follow up studies on the effect of their upregulation. Media optimization showed that addition of YEPD to the media led to a slight reduction in yield, but an increase in the production flux of the last step in psilocybin biosynthesis by a factor of 17.
A summary of what we achieved in this project:
- Computed a production strategy of heterologous psilocybin production in A. niger using a GSM model, and employed various computational methods to improve theoretical psilocybin yield.
- Computed gene regulation targets
- Investigated co-factor swapping
- Media optimization
- Our project is organized with two folders in the main folder. All of our work and analysis of the GSM, ATCC1015 iJB1325, including the model, is found in the Analysis folder.
- The report lives in the Report.ipynb notebook, and contains links to the notebooks in the Analysis folder.
- Pictures used in the report are stored in the Pics folder.
- Appendix is found in the Analysis folder.