Overripening and increased temperature: Alternative strategies to enhance peptide production and bioactivity in salt-reduced boneless cured Iberian hams

• Study shows how overripening and heat boost bioactivity in Iberian hams. • Overripening to 42% weight loss at 30 °C enhances ACE-I and antioxidant activities. • More bioactive peptides formed in hams overripened at higher temperatures. • Overripened hams may help regulate hypertension and oxidative stress. • Findings reveal new ways to develop healthier Iberian ham-based products. Iberian ham is renowned for its unique flavour and artisanal tradition, but its potential as a source of bioactive compounds remains underexplored. This study presents an innovative view of how overripening and controlled temperature increase can significantly improve the production and diversity of bioactive peptides in salt-reduced cured Iberian hams. Forty-eight hams were divided into three groups: traditionally cured, overripened with 42 % weight loss at 30 °C, and overripened with 42 % weight loss at 36°. Comparing them, our results reveal significant advances in proteolysis, generating peptides with potent antioxidant and angiotensin converting enzyme-I (ACE-I) inhibitory activities. In particular, hams cured at 30 °C showed superior bioactive profiles, reaching the lowest IC50 values for ACE-I activity, while those processed at 36 °C exhibited the highest diversity, identifying 1,313 unique peptides by advanced nano-liquid chromatography-mass spectrometry. In terms of antioxidant activity, overripeness significantly affected (p ≤ 0.05 ) antioxidant activity, with both overripened hams showing the highest antioxidant activity. These findings redefine the role of Iberian ham as a functional food with potential applications in cardiovascular and metabolic health management, while maintaining the authenticity and quality of traditional Iberian ham.