Studies enabled by metabolic models of different species of microalgae have become significant since they allow us to understand changes in their metabolism and physiological stages. The most used method to study cell metabolism is FBA, which commonly focuses on optimizing a single objective function. However, recent studies have brought attention to the exploration of simultaneous optimization of multiple objectives. Such strategies have found application in optimizing biomass and several other bioproducts of interest; they usually use approaches such as multi-level models or enumerations schemes. This work proposes an alternative in silico multiobjective model based on an evolutionary algorithm that offers a broader approximation of the Pareto frontier, allowing a better angle for decision making in metabolic engineering. The proposed strategy is validated on a reduced metabolic network of the microalgae Chlamydomonas reinhardtii while optimizing for the production of protein, carbohydrates, and CO${}_2$ uptake. The results from the conducted experimental design show a favorable difference in the number of solutions achieved compared to a classic tool solving FBA.

Flux Balance Analysis (FBA) is a constraint-based method that is commonly used to guide metabolites through restricting pathways that often involve conditions such as anaplerotic cycles like Calvin, reversible or irreversible reactions, and nodes where metabolic pathways branch. The method can identify the best conditions for one course but fails when dealing with the pathways of multiple metabolites of interest. Recent studies on metabolism consider it more natural to optimize several metabolites simultaneously rather than just one; moreover, they point out the use of metaheuristics as an attractive alternative that extends FBA to tackle multiple objectives. However, the literature also warns that the use of such techniques must not be wild. Instead, it must be subject to careful fine-tuning and selection processes to achieve the desired results. This work analyses the impact on the quality of the pathways built using the NSGAII and MOEA/D algorithms and several novel optimization models; it conducts a study on two case studies, the pigment biosynthesis and the node in glutamate metabolism of the microalgae Chlorella vulgaris, under three culture conditions (autotrophic, heterotrophic, and mixotrophic) while optimizing for three metabolic intermediaries as independent objective functions simultaneously. The results show varying performances between NSGAII and MOEA/D, demonstrating that the selection of an optimization model can greatly affect predicted phenotypes.