The study on preparation and functional properties of Spirulina active peptides is of significant importance for its deep processing.In this research, different proteases were employed to hydrolyze Spirulina proteins to screen and prepare active peptides with multiple biological activities.The results demonstrated that the degree of hydrolysis of Spirulina peptides (SPs) enzymatically hydrolyzed by flavourzyme reached 48.65%.Furthermore, the DPPH and ABTS free radical scavenging rates were 86.18% and 80.07%, respectively.The in vitro activation rate of alcohol dehydrogenase (ADH) was 126.80%, while the proportion of small molecule peptides (<1 kDa) accounted for 96.02%.Compared with other enzymes, flavor protease exhibited significant advantages in hydrolyzing Spirulina protein.Subsequently, an early alcoholic liver disease model of mice was established to evaluate the in vivo detoxification and hepatoprotective activities of SPs.It was found that the activities of ADH and aldehyde dehydrogenase (ALDH) in mice increased by 57.62% and 90.47% respectively, while aspartate transaminase (AST) and alanine transaminase (ALT) activities decreased by 40.27% and 43.63% respectively.Finally, the amino acid sequences of SPs were identified by ultra-high performance liquid chromatography-tandem linear ion trap/electrostatic field orbitrap high-resolution mass spectrometry, and 9 672 peptide segments were obtained by comparison with the Spirulina protein database. With the aid of computer-assisted simulation screening, 13 active peptide sequences with potential antioxidant activity were ultimately obtained.Four novel peptide segments with high ADH activation rates and dual antioxidant activity were discovered, among which HPGIP exhibited an ADH activation rate of 176.06%, along with DPPH and ABTS free radical scavenging rates of 93.36% and 87.12% respectively.This study provides valuable insights and references for the deep processing of Spirulina and the development of functional foods targeting alcohol detoxification and hepatoprotective effects.
[1] WILLIAMS R, AITHAL G, ALEXANDER G J, et al. Unacceptable failures: The final report of the Lancet Commission into liver disease in the UK[J]. The Lancet, 2020, 395(10219):226-239.
[2] 庞宇辰,郑俊侨,张思璐,等.解酒护肝胶囊对大鼠酒精性肝损伤保护作用研究[J]. 广东药科大学学报, 2024, 40(2):1-6.
PANG Y C, ZHENG J Q, ZHANG S L, et al. Protection effect of alcoholic liver injury by Jiejiu Hugan Capsules in rats [J]. Journal of Guangdong Pharmaceutical University, 2024, 40(2):1-6.
[3] YOKOYAMA H, BARAONA E, LIEBER C S. Upstream structure of human ADH7 gene and the organ distribution of its expression[J]. Biochemical and Biophysical Research Communications, 1995, 216(1):216-222.
[4] CENI E, MELLO T, GALLI A. Pathogenesis of alcoholic liver disease: Role of oxidative metabolism[J]. World Journal of Gastroenterology, 2014, 20(47):17756-17772.
[5] 杨胜涛.两种海藻多糖抑制肿瘤转移和血管新生作用及机制研究[D]. 湛江:广东海洋大学,2021.
YANG S T. Study on the effect and mechanism of two kinds of seaweed polysaccharide in inhibiting tumormetastasis and angiogenesis [D]. Zhanjiang: Guangdong Ocean University, 2021.
[6] PEACOCK A, LEUNG J, LARNEY S, et al. Global statistics on alcohol, tobacco and illicit drug use: 2017 status report[J]. Addiction, 2018, 113(10):1905-1926.
[7] SREELEKSHMI P J, DEVIKA V, AISWARYA L S, et al. Recent advances in bioactive peptides as functional food for Health Promotions and medicinal applications[J]. Protein & Peptide Letters, 2023, 30(8):626-639.
[8] GIORDANO M, LUONGO G, DAVINELLI S, et al. Silybum marianum: Not just silymarin and flavonolignans [J]. Records of Natural Products, 2021, 15(4):243-253.
[9] XIAO C Q, ZHAO M M, ZHOU F B, et al. Data on bioactive peptides derived from chicken hydrolysate with potential alcohol dehydrogenase stabilizing activity and in silico analysis of their potential activity and applicability[J]. Data in Brief, 2020, 29:105163.
[10] 刘文颖,冯晓文,李国明,等.牡蛎低聚肽的结构表征及体外抗氧化作用[J].中国食品学报,2021,21(12):261-269.
LIU W Y, FENG X W, LI G M, et al. Structure characterization and antioxidant effects in vitro of oyster oligopeptides [J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(12):261-269.
[11] 温庆仕,魏荷芬,周精卫,等.一种高纯度小分子解酒护肝肽的制备[J].食品与发酵工业,2023,49(7):174-180.
WEN Q S, WEI H F, ZHOU J W, et al. Preparation of a high purity small molecule anti-alcoholic liver protecting peptide [J]. Food and Fermentation Industries, 2023, 49(7):174-180.
[12] 赵谋明,马梅,苏国万,等.具有醒酒活性的玉米肽的制备、富集和鉴定[J].中国食品学报,2020,20(9):86-94.
ZHAO M M, MA M, SU G W, et al. Preparation, Enrichment and Identification of Corn Peptides with Facilitating Alcohol Metabolism Activity [J]. Journal of Chinese Institute of Food Science and Technology, 2020 ,20(9):86-94.
[13] ROMAY C, GONZALEZ R. Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals[J]. Journal of Pharmacy and Pharmacology, 2000, 52(4):367-368.
[14] HOSEINI S M, KHOSRAVI-DARANI K, MOZAFARI M R. Nutritional and medical applications of Spirulina microalgae[J]. Mini-Reviews in Medicinal Chemistry, 2013, 13(8):1231-1237.
[15] REDDY M C, SUBHASHINI J, MAHIPAL S V K, et al. C-Phycocyanin, a selective cyclooxygenase-2 inhibitor, induces apoptosis in lipopolysaccharide-stimulated RAW264.7 macrophages[J]. Biochemical and Biophysical Research Communications, 2003, 304(2):385-392.
[16] TONG X Y, PRASANNA G, ZHANG N, et al. Spectroscopic and molecular docking studies on the interaction of phycocyanobilin with peptide moieties of C-phycocyanin[J]. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2020, 236:118316.
[17] ZAN R, ZHU L, WU G C, et al. Identification of novel peptides with alcohol dehydrogenase (ADH) activating ability in chickpea protein hydrolysates[J]. Foods, 2023, 12(8):1574.
[18] 冯杨梦晓,任青兮,刘双平,等.海洋蛋白源解酒护肝水解物的筛选制备及功效评价[J].食品与发酵工业,2025,51(3):1-11.
FENG Y M X, REN Q X, LIU S P, et al. Screening, preparation, and efficacy evaluation of hydrolysates from marine protein sources for sobering up and liver protection [J]. Food and Fermentation Industries, 2025, 51(3):1-11.
[19] BERTOLA A, MATHEWS S, KI S H, et al. Mouse model of chronic and binge ethanol feeding (the NIAAA model)[J]. Nature Protocols, 2013, 8(3):627-637.
[20] MOONEY C, HASLAM N J, POLLASTRI G, et al. Towards the improved discovery and design of functional peptides: Common features of diverse classes permit generalized prediction of bioactivity[J]. PLoS One, 2012, 7(10): e45012.
[21] OLSEN T H, YESILTAS B, MARIN F I, et al. AnOxPePred: Using deep learning for the prediction of antioxidative properties of peptides[J]. Scientific Reports, 2020, 10:21471.
[22] WILKINS M R, GASTEIGER E, BAIROCH A, et al. Protein Identification and Analysis Tools in the ExPASy Server[M]. 2-D Proteome Analysis Protocols. New Jersey: Humana Press, 2003:531-552.
[23] MORAIS H A, SILVE M P C, SILVA V D M, et al. Correlation between the degree of hydrolysis and the peptide profile of whey protein concentrate hydrolysates: Effect of the enzyme type and reaction time[J]. American Journal of Food Technology, 2012, 8(1):1-16.
[24] TRAN H C, LE H A T, LE T T, et al. Effects of enzyme types and extraction conditions on protein recovery and antioxidant properties of hydrolysed proteins derived from defatted Lemna minor[J]. Applied Science and Engineering Progress, 2021,5(3):360-369.
[25] MARTINEZ-HURTADO J, CALO-FERNANDEZ B, VAZQUEZ-PADIN J. Preventing and mitigating alcohol toxicity: A review on protective substances[J]. Beverages, 2018, 4(2):39.
