Aunchalee WongaemOnrapak ReamtongPiroonporn SrimongkolPapassara SangtanooTanatorn SaisavoeyAphichart KarnchanatatChulalongkorn UniversityMahidol University2020-08-252020-08-252020-01-01Journal of Food Science and Technology. (2020)09758402002211552-s2.0-85086838098https://repository.li.mahidol.ac.th/handle/20.500.14594/57623© 2020, Association of Food Scientists & Technologists (India). 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. Graphic abstract: [Figure not available: see fulltext.].Mahidol UniversityAgricultural and Biological SciencesArticleSCOPUS10.1007/s13197-020-04582-4