Abstract

Myrosinase is essential for sulforaphane formation; however, its extraction from plant tissues yields low and highly variable recoveries, highlighting the need for recombinant production. Despite its relevance, knowledge on how to optimize recombinant myrosinase production in Saccharomyces cerevisiae remains limited. In this study, the cultivation kinetics of S. cerevisiae MGY70-myr were investigated in controlled bioreactor cultivations by combining experimental data with kinetic modeling and sensitivity analysis. Among the evaluated growth models, the Teissier model was identified as the most suitable for describing the biphasic behavior of the system. During the induction phase, high protein yields were achieved without evidence of physiological limitation. Sensitivity analysis revealed that biomass formation during the preculture phase was mainly governed by the maximum specific growth rate, whereas recombinant protein production during induction was strongly influenced by kinetic parameters associated with growth and non-growth-associated synthesis. Operational conditions exerted a decisive effect on process performance. Increased aeration and agitation enhanced biomass yield and productivity, whereas the accumulation of organic acids negatively affected metabolic efficiency and recombinant protein formation. Overall, this study identifies key kinetic parameters and operational conditions enabling process intensification of recombinant myrosinase production in yeast, providing a quantitative basis for understanding and optimizing bioreactor operation.