The effect of germination in combination with Alcalase hydrolysis of Brazilian soybean cultivar BRS 133 on the production of soybean flours with bioactive peptides as modulators of oxidative stress and markers of inflammation was monitored. The electrophoretic profile showed a weak protein breakdown during germination. However, a strong breakdown of the proteins can be observed after the first hour of hydrolysis with Alcalase. MALDI-TOF-MS analysis of the protein extracts showed differences in the intensity and profile of peptide mass fingerprint due to germination and hydrolysis. Germinated flour showed higher soluble protein concentration and antioxidant capacity. All soybean protein extracts and protein hydrolysates produced (G0, G18 and G72) showed a significant (p<0.05) inhibition on inflammatory markers such as nitric oxide (20.5-69.3%), iNOS (22.8-93.6%), PGE(2) (64.0-88.3%), COX-2 (36.2-76.7%), and TNF-α (93.9-99.5%) in LPS-induced RAW 264.7 macrophages. However, protein extracts of flours with 18h of germination were more potent in inhibiting pro-inflammatory responses when compared to 72h. It can be concluded that a combination of 72h of soybean BRS 133 germination and 1h Alcalase hydrolysis resulted in the formation of bioactive compounds with more potent antioxidant activity, and improvement in the reduction of some of the markers of inflammation.