Enzymatic hydrolysis preparation and activity analysis of antioxidant peptides derived from walnut dregs

Authors

NI Ce, School of Food Science and Bioengineering, Changsha University of Science ＆ Technology, Changsha, Hunan 410114, China
CAO Tianhong, School of Food Science and Bioengineering, Changsha University of Science ＆ Technology, Changsha, Hunan 410114, China
CHEN Min, School of Food Science and Bioengineering, Changsha University of Science ＆ Technology, Changsha, Hunan 410114, China;School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong 510006, China
YU Xun, School of Food Science and Bioengineering, Changsha University of Science ＆ Technology, Changsha, Hunan 410114, China
WU Hao, School of Food Science and Bioengineering, Changsha University of Science ＆ Technology, Changsha, Hunan 410114, China
WEN Li, School of Food Science and Bioengineering, Changsha University of Science ＆ Technology, Changsha, Hunan 410114, ChinaFollow

Corresponding Author(s)

文李（1971—），女，长沙理工大学教授，博士生导师，博士。E-mail: wl@csust.edu.cn

Abstract

Objective: This study focused on the identification and activity analysis of antioxidant active peptides in walnut dregs protein hydrolysates. Methods: Protein was extracted from walnut meal by fractionation method, and five kinds of proteolytic enzymes, including trypsin, papain, neutral protease, alkaline protease and flavor protease, were used to hydrolyze the proteins. The antioxidant activity of trypsin hydrolysate （TH）, papain hydrolysate （PH）, dispase hydrolysate （DH）, alcalase hydrolysate （AH） and flavourzyme hydrolysate （FPH） were evaluated and compared. The in vitro antioxidant activity of different concentrations of AH was determined by the detection indexes of DPPH free radical scavenging rate, ABTS free radical scavenging rate and hydroxyl free radical scavenging rate. The sequences of all peptides in AH were identified by LC-MS/MS technology, and the potential antioxidant active peptides of walnut were screened by PeptideRanker, BIOPEP-UWM database and molecular docking tool AutoDock1.5.6, and the two antioxidant active peptides were obtained. The above two peptides were simulated with Keap1 protein, and the antioxidant activity of the two antioxidant peptides was analyzed by using the zebrafish embryo oxidative damage model. Results: The optimal scavenging rates of AH with better antioxidant activity among the five enzymatic hydrolysates were （71.56±0.75）%, （60.63±0.45）% and （98.63±0.06）%, respectively. LC-MS/MS was identified by computer analysis to obtain two active ALWPF and PLRWPF peptides, both of which could bind to the active sites of Keap1 protein, with binding energies of -9.2 and -9.6 kJ/mol, respectively. The safe concentrations of zebrafish embryos treated with ALWPF and PLRWPF peptides ranged from 0～50 μg/mL and 0～30 μg/mL, respectively. Conclusion: Two antioxidant peptides were obtained from walnut meal, and the antioxidant effects of them were verified by zebrafish embryo model. The results of this study can provide a basis for the in-depth utilization of walnut protein.

Publication Date

7-22-2024

First Page

51

Last Page

61

DOI

10.13652/j.spjx.1003.5788.2024.60028

Recommended Citation

Ce, NI; Tianhong, CAO; Min, CHEN; Xun, YU; Hao, WU; and Li, WEN (2024) "Enzymatic hydrolysis preparation and activity analysis of antioxidant peptides derived from walnut dregs," Food and Machinery: Vol. 40: Iss. 5, Article 7.
DOI: 10.13652/j.spjx.1003.5788.2024.60028
Available at: https://www.ifoodmm.cn/journal/vol40/iss5/7

References

[1] 李明娟, 张雅媛, 游向荣, 等. 不同干燥技术对核桃粕蛋白粉品质特性及微观结构的影响[J]. 食品科学, 2021, 42（5）: 92-98. LI M J, ZHANG Y Y, YOU X R, et al. Effect of different drying technologies on the quality characteristics and microstructure of walnut meal protein powder[J]. Food Science, 2021, 42（5）: 92-98.
[2] QU T M, HE S W, NI C, et al. In vitro anti-inflammatory activity of three peptides derived from the byproduct of rice processing[J]. Plant Foods for Human Nutrition, 2022, 77（2）: 172-180.
[3] WEN L, HUANG L, LI Y W, et al. New peptides with immunomodulatory activity identified from rice proteins through peptidomic and in silico analysis[J]. Food Chemistry, 2021, 364: 130357.
[4] 文李, ISAAC N, 刘步青, 等. 食源性免疫活性肽的筛选策略及作用机制研究进展[J]. 食品与机械, 2020, 36（2）: 7-11, 37. WEN L, ISSAC N, LIU B Q, et al. Progresses of the screening strategies and the effect mechanism researches of food-derived immunopeptides[J]. Food & Machinery, 2020, 36（2）: 7-11, 37.
[5] LIU C L, GUO Y, ZHAO F R, et al. Potential mechanisms mediating the protective effects of a peptide from walnut （Juglans mandshurica Maxim.） against hydrogen peroxide induced neurotoxicity in PC12 cells[J]. Food & Function, 2019, 10（6）: 3 491-3 501.
[6] MAO R X, WU L, ZHU N, et al. Immunomodulatory effects of walnut （Juglans regia L.） oligopeptides on innate and adaptive immune responses in mice[J]. Journal of Functional Foods, 2020, 73: 104068.
[7] MOGHADAM M, SALAMI M, MOHAMMADIAN M, et al. Walnut protein-curcumin complexes: Fabrication, structural characterization, antioxidant properties, and in vitro anticancer activity[J]. Journal of Food Measurement and Characterization, 2020, 14（2）: 876-885.
[8] ABD EL-SALAM M H, EL-SHIBINY S. Preparation, properties, and uses of enzymatic milk protein hydrolysates[J]. Critical Reviews in Food Science and Nutrition, 2017, 57（6）: 1 119-1 132.
[9] JIA Q, YUAN J F, LIU H P, et al. Purification and identification of dual-enzyme hydrolysates obtained from defatted walnut and its antioxidant effects ond-galactose-induced aging mice[J]. Journal of Food Measurement and Characterization, 2020, 15: 1 034-1 043.
[10] WANG J Y, LU S L, LI R T, et al. Identification and characterization of antioxidant peptides from Chinese dry-cured mutton ham[J]. Journal of the Science of Food and Agriculture, 2020, 100（3）: 1 246-1 255.
[11] YU Z P, CAO Y X, KAN R T, et al. Identification of egg protein-derived peptides as xanthine oxidase inhibitors: Virtual hydrolysis, molecular docking, and in vitro activity evaluation[J]. Food Science and Human Wellness, 2022, 11（6）: 1 591-1 597.
[12] TU W J, WANG H, LI S, et al. The anti-inflammatory and anti-oxidant mechanisms of the Keap1/Nrf2/ARE signaling pathway in chronic diseases[J]. Aging and Disease, 2019, 10（3）: 637-651.
[13] KO J Y, KIM E A, LEE J H, et al. Protective effect of aquacultured flounder fish-derived peptide against oxidative stress in zebrafish[J]. Fish & Shellfish Immunology, 2014, 36（1）: 320-323.
[14] WANG Y Y, WANG C Y, WANG S T, et al.Physicochemical properties and antioxidant activities of tree peony （Paeonia suffruticosa Andr.） seed protein hydrolysates obtained with different proteases[J]. Food Chemistry, 2021, 345: 128765.
[15] WANG J, XIE Y J, LUAN Y Y, et al. Identification and dipeptidyl peptidase Ⅳ （DPP-Ⅳ） inhibitory activity verification of peptides from mouse lymphocytes[J]. Food Science and Human Wellness, 2022, 11（6）: 1 515-1 526.
[16] 张迎阳, 董亚云, 邹平, 等. 橄榄仁抗氧化肽的分离鉴定及分子对接[J]. 食品与发酵工业, 2024, 50（5）: 189-198. ZHANG Y Y, DONG Y Y, ZOU P, et al. Isolation,identification and molecluar docking of antioxidant peptides from olive kernel[J]. Food and Fermentation Industries, 2024, 50（5）: 189-198.
[17] 邹娅雪, 付晓婷, 段德麟, 等. 利用斑马鱼模型研究琼胶寡糖抗氧化机制[J]. 食品工业科技, 2019, 40（4）: 286-291, 298. ZOU Y X, FU X T, DUAN D L, et al. Antioxidant activities of agaro-oligosaccharides in AAPH-induced zebrafish model[J]. Science and Technology of Food Industry, 2019, 40（4）: 286-291, 298.
[18] 沈金鹏, 王珂雯, 黄潘钿, 等. 珍珠贝水解肽的制备、氨基酸组成及抗炎活性[J]. 食品与机械, 2023, 39（2）: 132-139, 206. SHEN J P, WANG K W, HUANG P D, et al. Preparation,amino acid composition and anti-inflammatory activity of hydrolyzed peptides from Pinctada martensii[J]. Food & Machinery, 2023, 39（2）: 132-139, 206.
[19] GU M, CHEN H P, ZHAO M M, et al. Identification of antioxidant peptides released from defatted walnut （Juglans Sigillata Dode） meal proteins with pancreatin[J]. LWT-Food Science and Technology, 2015, 60（1）: 213-220.
[20] 李丽, 阮金兰, 钱伟亮, 等. 多指标评价核桃蛋白及多肽的抗氧化活性[J]. 食品研究与开发, 2017, 38（3）: 1-4. LI L, RUAN J L, QIAN W L, et al. Studies on the antioxidant activity of walnut protein and polypeptide by multi-index[J]. Food Research and Development, 2017, 38（3）: 1-4.
[21] 罗燕, 和兴萍, 李雪, 等. 三种蛋白酶酶解核桃饼粕的抗氧化活性研究[J]. 中国酿造, 2017, 36（5）: 170-174. LUO Y, HE X P, LI X, et al. Study on antioxidant activity of three kinds of protease for enzymatic hydrolysis of walnut cake[J]. China Brewing, 2017, 36（5）: 170-174.
[22] 贾靖霖, 陆健康, 汪莉莉, 等. 核桃多肽体外抗氧化活性研究[J]. 中国酿造, 2014, 33（5）: 109-112. JIA J L, LU J K, WANG L L, et al. Study on antioxidant activity of walnut polypeptide in vitro[J]. China Brewing, 2014, 33（5）: 109-112.
[23] WANG J Y, LIU J L, JOHN A T, et al. Structure identification of walnut peptides and evaluation of cellular antioxidant activity[J]. Food Chemistry, 2022, 388: 132943.
[24] 贺舒雯, 朱豪杰, 韩鹏薇, 等. 虾壳活性肽对斑马鱼氧化应激损伤的保护作用[J]. 食品与机械, 2023, 39（9）: 140-147. HE S W, ZHU H J, HAN P W, et al. Protective effects of Procambarus clarkii shell bioactive peptides on oxidative stress injury of zebrafish （Danio rerio）[J]. Food & Machinery, 2023, 39（9）: 140-147.
[25] 郭勇, 秦汉雄, 魏贞, 等. 长白山核桃源五肽对过氧化氢诱导PC12细胞氧化损伤的保护作用及机理[J]. 食品科学, 2019, 40（13）: 143-149. GUO Y, QIN H Y, WEI Z, et al. Cytoprotective effect and underlying mechanism of pentapeptide derived from manchurian walnut（Juglans mandshurica Maxim） meal protein on PC12 cells against H₂O₂-induced damage[J]. Food Science, 2019, 40（13）: 143-149.
[26] ZHU X L, XIAO X J, WANG S, et al. Rosendaal linear interpolation method appraising of time in therapeutic range in patients with 12-week follow-up interval after mechanical heart valve replacement[J]. Front Cardiovasc Med, 2022, 9: 925571.
[27] LI Y, WANG R X, LI Y Q, et al. Protective effects of tree peony seed protein hydrolysate on Cd-inducedoxidative damage, inflammation and apoptosis in zebrafish embryos[J]. Fish & Shellfish Immunology, 2022, 126: 292-302.