{Reference Type}: Journal Article {Title}: Factors impacting the microbial production of eicosapentaenoic acid. {Author}: Sousa S;Carvalho AP;Gomes AM; {Journal}: Appl Microbiol Biotechnol {Volume}: 108 {Issue}: 1 {Year}: 2024 Jun 11 {Factor}: 5.56 {DOI}: 10.1007/s00253-024-13209-z {Abstract}: The increasing applications for eicosapentaenoic acid (EPA) and the potential shortfall in supply due to sustainability and contamination issues related with its conventional sources (i.e., fish oils; seafood) led to an extensive search for alternative and sustainable sources, as well as production processes. The present mini-review covers all the steps involved in the production of EPA from microorganisms, with a deeper focus on microalgae. From production systems to downstream processing, the most important achievements within each area are briefly highlighted. Comparative tables of methodologies are also provided, as well as additional references of recent reviews, so that readers may deepen their knowledge in the different issues addressed. KEY POINTS: • Microorganisms are more sustainable alternative sources of EPA than fish. • Due to the costly separation from DHA, species that produce only EPA are preferable. • EPA production can be optimised using non-genetic and genetic tailoring engineering.