# Rice Bran-Derived Peptides with Antioxidant Activity: Effects of Enzymatic Hydrolysis Using Bacillus licheniformis and α-Chymotrypsin

## TL;DR

This study explores how enzymes can break down rice bran proteins into small peptides with strong antioxidant properties.

## Contribution

The novel contribution is identifying the <1 kDa peptide fraction from α-chymotrypsin hydrolysis as having the strongest antioxidant activity.

## Key findings

- The <1 kDa peptide fraction from α-chymotrypsin hydrolysis showed the lowest EC50 values for ABTS and DPPH.
- This fraction also had the highest inhibition of metal chelating activity and linoleic peroxidation.
- Enzymatic hydrolysis increased the total phenolic content compared to native rice bran protein.

## Abstract

Rice bran, a nutrient-rich by-product of rice milling, is an underutilized resource in sustainable crop utilization. This study aimed to investigate the characteristics, total phenolic content, and antioxidant activities of rice bran protein hydrolysates (RBPHs) produced using proteases from Bacillus licheniformis (RBPH-B) and α-chymotrypsin (RBPH-C), along with their protein fractions (F1; >100 kDa, F2; 10–100 kDa, F3; 1–10 kDa, F4; <1 kDa). Molecular weight, color, surface hydrophobicity, secondary structure, total phenolic content, and antioxidant activities of the hydrolysates were assessed. Both enzymatic hydrolysis and ultrafiltration reduced molecular weight and surface hydrophobicity, enhanced lightness, and increased α-helix content. Among all samples, the <1 kDa peptide fraction derived from α-chymotrypsin hydrolysis (RBPH-C-F4) exhibited the strongest antioxidant activity, with the lowest EC50 values for ABTS (0.94 mg/mL) and DPPH (210 µg/mL), as well as the highest inhibition of metal chelating activity (1.35 mmol EDTA/g sample) and linoleic peroxidation (90.62%). Enzymatic hydrolysis enhanced total phenolic content compared with native rice bran protein. These findings highlight the potential of rice bran-derived peptides as antioxidant candidates and indicate that further validation in food systems is required.