Nannochloropsis oceanica is a microalga with relevant protein content, making it a potential source of bioactive peptides. Furthermore, it is also rich in fatty acids, with a special focus on eicosapentaenoic acid, an omega‐3 fatty acid mainly obtained from marine animal sources, with high importance for human health.N. oceanica has a rigid cell wall constraining protein extraction, thus hydrolyzing it may help increase its components' extractability. Therefore, a Box‐Behnken experimental design was carried out to optimize the hydrolysis.The hydrolysate A showed 67±0.7% of protein, antioxidant activity of 1166±63.7 μmol TE/g of protein and an ACE inhibition with an IC50 of 379 μg protein/mL. The hydrolysate B showed 60±1.8% of protein, antioxidant activity of 775±13.0 μmol TE/g of protein and an ACE inhibition with an IC50 of 239 μg protein/mL. The by‐product showed higher yields of total fatty acids when compared to “raw” microalgae, being 5.22 and 1%, respectively.The sustainable developed methodology led to the production of one fraction rich in bioactive peptides and another with interesting EPA content, both with value‐added properties with potential to be commercialized as ingredients for different industrial applications, such as functional food, supplements or cosmetic formulations.This article is protected by copyright. All rights reserved