Enzymatic optimization for enhancing earthworm peptide yields and biological activities on antioxidant, cytotoxic against HT-29 and HepG2 cancer cells, and antimigration.
- 1Program in Biotechnology, Faculty of Science, Maejo University, Chiang Mai, Thailand.
Abstract
The purposes of this study were to investigate the antioxidant, cytotoxic against HT-29 and HepG2 cancer cells, and antimigratory activities of peptide fractions derived from Amynthas arenulus earthworm. Earthworm proteins were hydrolyzed with alcalase using response surface methodology. The optimal conditions resulted in a maximum degree of hydrolysis of 61.25% at an enzyme-to-substrate ratio of 1.5% (v/v) and a hydrolysis time of 2 h. Following simulated gastrointestinal digestion, gastric and intestinal recoveries of the peptides were 56.83% and 90.10%, respectively. The 10-30 kDa peptide fraction exhibited antioxidant activity, with estimated IC₅₀ values of 3.18 and 0.79 mg/mL for DPPH and ABTS assays, respectively. FRAP activity was also exhibited. The 10-30 kDa peptide fraction showed antiproliferative activity against HT-29 and HepG2 cells, with IC₅₀ values of 0.634 and 0.761 mg/mL, respectively, whereas it was not cytotoxic against non-cancerous cells of L929 fibroblast cells. This peptide fraction was trigger apoptosis-associated cellular changes observed using inverted light microscopy, AO/PI and DAPI staining, DNA fragmentation, and scanning electron microscopy. Relative cell migration values in HT-29 and HepG2 cells were 67.34% and 70.29%, respectively which compared to 100% of control. As resultants, this peptide fraction might be considered for applied in medical, nutraceutical food, and bioprocessing industries.
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BibTeX
@article{wasunan2026enzymatic,
title = {{Enzymatic optimization for enhancing earthworm peptide yields and biological activities on antioxidant, cytotoxic against HT-29 and HepG2 cancer cells, and antimigration.}},
author = {Pimphan Wasunan and Mongkol Thirabunyanon},
journal = {Frontiers in bioengineering and biotechnology},
year = {2026},
month = oct,
doi = {10.3389/fbioe.2026.1957959},
url = {https://europepmc.org/article/MED/42948006},
note = {CC BY},
}
RIS
TY - JOUR
TI - Enzymatic optimization for enhancing earthworm peptide yields and biological activities on antioxidant, cytotoxic against HT-29 and HepG2 cancer cells, and antimigration.
AU - Pimphan Wasunan
AU - Mongkol Thirabunyanon
PY - 2026
DA - 2026/10/13
JO - Frontiers in bioengineering and biotechnology
DO - 10.3389/fbioe.2026.1957959
UR - https://europepmc.org/article/MED/42948006
AB - The purposes of this study were to investigate the antioxidant, cytotoxic against HT-29 and HepG2 cancer cells, and antimigratory activities of peptide fractions derived from Amynthas arenulus earthworm. Earthworm proteins were hydrolyzed with alcalase using response surface methodology. The optimal conditions resulted in a maximum degree of hydrolysis of 61.25% at an enzyme-to-substrate ratio of 1.5% (v/v) and a hydrolysis time of 2 h. Following simulated gastrointestinal digestion, gastric and intestinal recoveries of the peptides were 56.83% and 90.10%, respectively. The 10-30 kDa peptide fraction exhibited antioxidant activity, with estimated IC₅₀ values of 3.18 and 0.79 mg/mL for DPPH and ABTS assays, respectively. FRAP activity was also exhibited. The 10-30 kDa peptide fraction showed antiproliferative activity against HT-29 and HepG2 cells, with IC₅₀ values of 0.634 and 0.761 mg/mL, respectively, whereas it was not cytotoxic against non-cancerous cells of L929 fibroblast cells. This peptide fraction was trigger apoptosis-associated cellular changes observed using inverted light microscopy, AO/PI and DAPI staining, DNA fragmentation, and scanning electron microscopy. Relative cell migration values in HT-29 and HepG2 cells were 67.34% and 70.29%, respectively which compared to 100% of control. As resultants, this peptide fraction might be considered for applied in medical, nutraceutical food, and bioprocessing industries.
SN - 2296-4185
ER -