This project investigates the biosynthesis of isoamyl acetate using the yeast Saccharomyces cerevisiae, to optimize its enhanced production. Isoamyl Acetate, known as a banana flavor ester, is used in the food, beverage, pharmaceutical, and cosmetic industries. Our report begins with a literature review, outlining the metabolic pathway of isoamyl acetate synthesis and its status on the market. Product-related issues include low production rate, which we aim to solve through metabolic engineering strategies. Therefore, we applied MEMOTE comparison for 7 potential S. cerevisiae models to identify the most suitable model for the project. We conducted yields and media optimization analysis, phenotypic phase plane analysis, FSEOF, and co-factor-swapping optimization analysis to highlight the main bottlenecks of the process and find feasible solutions for the product. The DFBA showed how cell respond to applied environmental conditions in batch fermentation simulation.

This projects is organized in a following way. Report cointains core text description of the project covering introduction, problem definition, selection and assesment of excisting GSM, computer-aided cell factory engineering, discussion and conclusion parts with short statements, tables and figures. Main code with results is splited between 6 files related to different analyses.

1. 02 Model and pathway - Loading the model and analyzing the pathway and the target compound.
2. 03 Theoretical yields and media optimization - Yields and media optimization analysis.
3. 04 Phenotype phase plane - Phenotypic phase plane analysis on the baseline growth and progesterone production for identification cell response to changes in conditions.
4. 05 Gene target analysis - FSEOF analysis of the model with identification of targets for downregulation and upregulation.
5. 06 Co-factor swap - Co-factor-swapping targets analysis.
6. 07 Dynamic FBA - Dynamic flux analysis to identify cell response to set environmental conditions during fermentation.

In the folder Files reactions with manually changed co-factors can be found. Folder Models contains yeast models we used for MEMOTE analysis. We put figures that are used in the Report to the folder Pictures.