This study sought to assess the ideal conditions under which hydrolysate can be produced from the split gill mushroom proteins through the microbial protease, Alcalase. The research employed a central composite design and response surface methodology. Three specific parameters were varied for the purposes of the experimental process, while a fixed pH value of 8 was used in all cases. The variables were hydrolysis temperature (set as 45 °C, 50 °C, or 55 °C), hydrolysis time (set as 60 min, 120 min, or 180 min), and the ratio of enzyme to substrate (set as 2%, 4%, or 6% w/v). The variables under investigation exert a significant influence upon degree of hydrolysis (DH) in addition to 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging activity (p < 0.05). Fractionation of the hydrolysate was accomplished using molecular weight (MW) cut-off membranes, while the greatest radical-scavenging capability was observed in the < 0.65 kDa fraction. The MW < 0.65 kDa fraction underwent separation through RP-HPLC in order to create five sub-fractions. Among these, the greatest ABTS radical-scavenging capability was observed in the F5 sub-fraction, which was therefore chosen to undergo additional examination using quadrupole-time-of-flight-electron spin induction-mass spectrometry-based de novo peptide sequencing. Via this process it was possible to determine five antioxidant peptides. Furthermore, the MW < 0.65 kDa fraction was able to demonstrating cellular antioxidant activity in the context of a human intestinal cancer cell line (HT-29). The extent of this activity was shown to depend upon the concentration levels of the peptide.
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The authors would like to thank the Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, for their support and providing access to their facilities. We acknowledge the financial support from the Grant for Research: The Research Assistantship Fund, Faculty of Science, Chulalongkorn University (RAF_2561_010), The Center of Excellence on Medical Biotechnology (CEMB), S&T Postgraduate Education and Research Development Office (PERDO), Office of Higher Education Commission (OHEC), Thailand (SN-60-003-909), and the Ratchadapisek Sompoch Endowment Fund, Chulalongkorn University (R_016_2556), and the Ratchadapisek Sompoch Endowment Fund (2019), Chulalongkorn University (762008) for providing the financial support for this research. The authors were grateful to Dr. Robert Douglas John Butcher for reviewing this manuscript.
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Wongaem, A., Reamtong, O., Srimongkol, P. et al. Antioxidant properties of peptides obtained from the split gill mushroom (Schizophyllum commune). J Food Sci Technol 58, 680–691 (2021). https://doi.org/10.1007/s13197-020-04582-4
- Free radical scavenging activity
- Protein hydrolysate
- Response surface methodology
- Schizophyllum commune
- Split gill mushroom