Combinatorial optimization of pathway, process and media for the production of p-coumaric acid by Saccharomyces cerevisiae
Moreno Paz, S.; van der Hoek, R.; Eliana, E.; Martins dos Santos, V. A. P.; Schmitz, J.; Suarez-Diez, M.
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Microbial cell factories are instrumental in transitioning towards a sustainable bio-based economy, offering alternatives to conventional chemical processes. However, fulfilling their potential requires simultaneous screening for optimal media composition, process and genetic factors, acknowledging the complex interplay between the organisms genotype and its environment. This study employs statistical Design of Experiments (DoE) to systematically explore these relationships and optimize the production of p-coumaric acid (pCA) in Saccharomyces cerevisiae. Two rounds of fractional factorial designs were used to identify factors with a significant effect on pCA production, which resulted on a 168-fold improvement on pCA titer. Moreover, a significant interaction between the culture temperature and expression of ARO4 highlighted the importance of simultaneous process and strain optimization. The presented approach leverages the strengths of experimental design and statistical analysis and could be systematically applied during strain and bio-process design efforts to unlock the full potential of microbial cell factories.
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