Keywords

Haddock Skin Collagen, Trypsin, Peptides, Molecular Docking, Light-Damaged Hacat Cells

Abstract

Objective: To study the antioxidant and photodamage-inhibiting effects of tryptic hydrolyzate peptides from haddock (Melanogrammus aeglefinus) skin collagen, and to improve the reuse value of fish skin. Methods: Single factor experiment and response surface experimental design were used to explore the optimal enzymatic hydrolysis process of haddock skin collagen. The peptide sequence of the prepared enzymatic peptide was determined by matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF MS/MS). The CDOCKER module in Discovery Studio was used to conduct molecular docking of the obtained advantageous peptides with the antioxidant-related Kelch-like epichlorohydrin-related protein-1 (Keap1). After pretreating human epidermal keratinocytes (HaCaT) with polypeptides of different mass concentrations, medium-wave ultraviolet rays were used to induce HaCaT cells to construct a photodamage model. (glutathione peroxidase, GSH-Px) activity, malondialdehyde (MDA) content, and cell apoptosis and cycle to study the ability of enzymatic peptides to inhibit photodamage. Results: The degree of hydrolysis of haddock skin collagen was 78.33% under the conditions of trypsin mass fraction 1.23%, enzymatic hydrolysis temperature 52.95?¡æ, and enzymatic hydrolysis time 1.75?h. MALDI-TOF MS/MS determined that the dominant peptide sequences in the enzymatic hydrolyzate were V-IFFVTMGTP-R, L-SIVPV-R and L-MHT-T. Through molecular docking, it was found that V-IFFVTMGTP-R binds to Keap1 most stably, thus predicting its antioxidant effect. When the light-damaged HaCaT cells were treated with 250?mg/mL enzymatic solution, their SOD and GSH-Px activities were significantly increased compared with the model group (P<0.01), and the MDA content was extremely significantly reduced (P<0.01). Conclusion: Haddock skin collagen trypsin-enzymatic peptide has antioxidant activity and has a certain inhibitory effect on photodamage.   For further details regarding this article and its research, please do not hesitate to contact our team for assistance. Original research done by Si Kaixue, Zhou Jun, Cui Chenqian, Zhang Hongyan, Li Yanyan, Wang Chaoyang, Lu Chenyang, Su Xiurong, Dong Lisa, Han Jiaojiao  

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