Food and Fermentation Industries >

2025 , Vol. 51 >Issue 4: 145 - 153

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.038957

Effect of O-glycosylation on structure and antioxidant activity of Enterococcus faecium AS8 glycopeptides

  • LI Shuxun ,
  • BAI Ying
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  • (College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China)

Received date: 2024-02-25

  Revised date: 2024-04-15

  Online published: 2025-03-10

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Abstract

Glycosylation plays a crucial role in the activity and conformation of proteins/peptides.This study utilized the milk fermentation broth of Enterococcus faecium AS8 as a raw material for extracting glycopeptides.The extraction rate of crude glycopeptides was used as a key indicator, and the extraction process was optimized through single factor and response surface experiments.Subsequently, the glycopeptides were purified via gel column chromatography to obtain the AS8 glycopeptide (AGP).AGP was evaluated for purity using SDS-PAGE and HPGPC, followed by the removal of the O-sugar chain in AGP through the β-elimination method to obtain the deglycosylated AS8 glycopeptide (dAGP).AGP and dAGP were then analyzed using infrared spectroscopy, fluorescence spectroscopy, scanning electron microscopy, thermal stability, and antioxidant activity measurements to preliminarily investigate the impact of O-glycosylation on the structure and antioxidant activity of AGP.Results indicated that deglycosylation led to a significant reduction in the intensity of certain infrared absorption peaks of the AGP, as well as a decrease in the main secondary structure β-folding.Additionally, there was an increase in endogenous fluorescence intensity, a red-shift in the maximum emission wavelength, and a decrease in the maximum fluorescence intensity of the exogenous source.The surface hydrophobicity was also reduced, along with a decrease in thermal stability.The microstructure changed from smooth and compact to rough and porous.Furthermore, the scavenging capacity of ABTS cationic free radicals was enhanced, while the scavenging capacity of DPPH radicals and ·OH scavenging capacity decreased.Overall, these findings suggest that the removal of O-glycosyl chains alters the antioxidant activity of AGP and diminishes its structural stability.

Key words： glycopeptide; deglycosylation; structure; antioxidant activity

Cite this article

LI Shuxun , BAI Ying . Effect of O-glycosylation on structure and antioxidant activity of Enterococcus faecium AS8 glycopeptides[J]. Food and Fermentation Industries, 2025 , 51(4) : 145 -153 . DOI: 10.13995/j.cnki.11-1802/ts.038957

References

[1] ZHANG M, OU X Z, SHI H, et al.Isolation, structures and biological activities of medicinal glycoproteins from natural resources:A review[J].International Journal of Biological Macromolecules, 2023, 244:125406.
[2] CÓRDOVA-DÁVALOS L E, CERVANTES-GARCÍA D, BALLONA-ALBA M F, et al.Protective effect of glycomacropeptide on the inflammatory response of U937 macrophages[J].Foods, 2023, 12(7):1528.
[3] LUO H J, ZHANG Y K, WANG S Z, et al.Structural characterization and anti-oxidative activity for a glycopeptide from Ganoderma lucidum fruiting body[J].International Journal of Biological Macromolecules, 2024, 261(Pt 2):129793.
[4] MOREMEN K W, TIEMEYER M, NAIRN A V.Vertebrate protein glycosylation:Diversity, synthesis and function[J].Nature Reviews Molecular Cell Biology, 2012, 13(7):448-462.
[5] YANG A S, DENG H, ZU Q Q, et al.Structure characterization and IgE-binding of soybean 7S globulin after enzymatic deglycosylation[J].International Journal of Food Properties, 2018, 21(1):171-182.
[6] KAR B, VERMA P, DEN HAAN R, et al.Effect of N-linked glycosylation on the activity and stability of a β-glucosidase from Putranjiva roxburghii[J].International Journal of Biological Macromolecules, 2018, 112:490-498.
[7] 向媛嫄, 王文林, 宋海云, 等.去糖基化对水溶澳洲坚果糖肽结构和抗氧化性的影响[J].食品与发酵工业, 2021, 47(11):98-103.
XIANG Y Y, WANG W L, SONG H Y, et al.Effect of deglycosylation on the structure and antioxidant activity of water-soluble macadamia glycopeptide[J].Food and Fermentation Industries, 2021, 47(11):98-103.
[8] 刘立杰. 三株乳酸菌对小鼠肝脏抗氧化能力及肠道粘膜免疫的影响[D].呼和浩特:内蒙古农业大学, 2017.
LIU L J.Effects of three lactic acid bacteria on liver antioxidant capacity and intestinal mucosa immunity of mice[D].Hohhot:Inner Mongolia Agricultural University, 2017.
[9] 石雅彤, 白英.模拟消化对副干酪乳杆菌LX5和屎肠球菌AS8发酵液抗氧化活性的影响[J/OL].食品与发酵工业, 2023.https://doi.org/10.13995/j.cnki.11-1802/ts.036844.
SHI Y T, BAI Y.Effect of simulated digestion on antioxidant activity of Lactobacillus paracasei LX5 and Enterococcus faecalis AS8 fermentation broth[J/OL].Food and Fermentation Industries, 2023.https://doi.org/10.13995/j.cnki.11-1802/ts.036844.
[10] ALLEY W R Jr, MANN B F, NOVOTNY M V.High-sensitivity analytical approaches for the structural characterization of glycoproteins[J].Chemical Reviews, 2013, 113(4):2668-2732.
[11] 李成, 王承健, 晋万军, 等.银杏种子糖蛋白的 SDS-PAGE 分离及N-糖链的质谱分析[J].高等学校化学学报, 2019, 40(1):69-75.
LI C, WANG C J, JIN W J, et al.Mass spectrometric analysis of N-glycans of glycoprotein separated by SDS-PAGE gel from ginkgo seed[J].Chemical Journal of Chinese Universities, 2019, 40(1):69-75.
[12] LI T T, WU C E, MENG X Y, et al.Structural characterization and antioxidant activity of a glycoprotein isolated from Camellia oleifera Abel seeds against D-galactose-induced oxidative stress in mice[J].Journal of Functional Foods, 2020, 64:103594.
[13] 邓雯, 廖雅如, 黄丽衡, 等.异黄酮与β-伴大豆球蛋白的相互作用及其对蛋白结构和潜在致敏性的影响[J/OL].食品科学, 2024.http://kns.cnki.net/kcms/detail/11.2206.TS.20240126.1101.002.html.
DENG W, LIAO Y R, HUANG L H, et al.Interaction of isoflavones with β-conglycinin and its effect on the structure of the complex and potential allergenicity[J/OL].Food Science, 2024.http://kns.cnki.net/kcms/detail/11.2206.TS.20240126.1101.002.html.
[14] ZHANG H, ZOU P, ZHAO H T, et al.Isolation, purification, structure and antioxidant activity of polysaccharide from pinecones of Pinus koraiensis[J].Carbohydrate Polymers, 2021, 251:117078.
[15] 李宁宁, 王正荣, 孙春晓, 等.牛蒡多糖对发酵乳品质和抗氧化性的影响[J].食品与发酵工业, 2019, 45(10):97-103.
LI N N, WANG Z R, SUN C X, et al.Burdock polysaccharides enhanced the quality and antioxidative activity of fermented milk[J].Food and Fermentation Industries, 2019, 45(10):97-103.
[16] ZHANG X, WANG C, HAN Q, et al.Highly sialylated mucin-type glycopeptide from porcine intestinal mucosa after heparin extraction:O-glycan profiling and immunological activity evaluation[J].Glycoconjugate Journal, 2021, 38(5):527-537.
[17] LI Y, LIU B H, JIANG L Z, et al.Interaction of soybean protein isolate and phosphatidylcholine in nanoemulsions:A fluorescence analysis[J].Food Hydrocolloids, 2019, 87:814-829.
[18] JIN H, ZHAO Q S, FENG H Y, et al.Changes on the structural and physicochemical properties of conjugates prepared by the Maillard reaction of black bean protein isolates and glucose with ultrasound pretreatment[J].Polymers, 2019, 11(5):848.
[19] ZHANG H Q, HAN L W, SUN X M, et al.A glycoprotein from mountain cultivated ginseng:Insights into their chemical characteristics and intracellular antioxidant activity[J].International Journal of Biological Macromolecules, 2022, 217:761-774.
[20] WANG Y, ZOU T T, XIANG M H, et al.Purification and characterization of a soluble glycoprotein from garlic (Allium sativum) and its in vitro bioactivity[J].Preparative Biochemistry and Biotechnology, 2016, 46(7):709-716.
[21] ZENG H J, LIU Z, WANG Y P, et al.Studies on the anti-aging activity of a glycoprotein isolated from Fupenzi (Rubus chingii Hu.) and its regulation on klotho gene expression in mice kidney[J].International Journal of Biological Macromolecules, 2018, 119:470-476.
[22] GAO J Y, LI X, JIA S T, et al.Structural characterization and antioxidant activity of a glycoprotein isolated from shiitake mushrooms[J].Food Bioscience, 2023, 53:102608.
[23] SHENG L, HE Z J, CHEN J H, et al.The impact of N-glycosylation on conformation and stability of immunoglobulin Y from egg yolk[J].International Journal of Biological Macromolecules, 2017, 96:129-136.
[24] 张永金, 胡艳红, 葛武鹏, 等.母乳、牛乳与主要小品种乳蛋白质组成及乳清蛋白二级结构比较[J].食品安全质量检测学报, 2022, 13(15):4779-4786.
ZHANG Y J, HU Y H, GE W P, et al.Comparative study on the composition of protein and secondary structure of whey protein in human milk, milk and main small varieties of milk[J].Journal of Food Safety and Quality, 2022, 13(15):4779-4786.
[25] YONG Y H, YAMAGUCHI S, MATSUMURA Y.Effects of enzymatic deamidation by protein-glutaminase on structure and functional properties of wheat gluten[J].Journal of Agricultural and Food Chemistry, 2006, 54(16):6034-6040.
[26] 李颖畅, 师丹华, 赵淞民, 等.超声波辅助没食子酸对鲈鱼肌原纤维蛋白结构特性的影响[J].中国食品学报, 2023, 23(7):88-98.
LI Y C, SHI D H, ZHAO S M, et al.Effect of gallic acid on structural properties of Lateolabrax japonicas myofibrillar protein under ultrasonic treatment[J].Journal of Chinese Institute of Food Science and Technology, 2023, 23(7):88-98.
[27] 王岸娜, 苏子豪, 吴立根, 等.O-糖链对猕猴桃糖蛋白抗氧化能力及构象的影响[J].河南工业大学学报(自然科学版), 2016, 37(2):79-85.
WANG A N, SU Z H, WU L G, et al.The impact of O-glycan on the antioxidant activity and secondary structure of glycoprotein extracted from kiwi fruit[J].Journal of Henan University of Technology (Natural Science Edition), 2016, 37(2):79-85.
[28] CHEN Y J, LIN Q L, WANG J Q, et al.Proteins, polysaccharides and their derivatives as macromolecular antioxidant supplements:A review of in vitro screening methods and strategies[J].International Journal of Biological Macromolecules, 2023, 224:958-971.
[29] 石嘉怿, 张太, 梁富强.体外模拟消化对大米谷蛋白结构及水解产物生物活性的影响[J].食品科学, 2021, 42(1):59-66.
SHI J Y, ZHANG T, LIANG F Q.Effect of in vitro simulated digestion on the structure of rice glutelin and the biological activity of hydrolysates[J].Food Science, 2021, 42(1):59-66.
[30] WU Y T, HUO Y F, XU L, et al.Purification, characterization and antioxidant activity of polysaccharides from Porphyra haitanensis[J].International Journal of Biological Macromolecules, 2020, 165:2116-2125.
[31] YANG R W, WANG J, LIN S Y, et al.In vitro antioxidant activities of the novel pentapeptides Ser-His-Glu-Cys-Asn and Leu-Pro-Phe-Ala-Met and the relationship between activity and peptide secondary structure[J].Journal of the Science of Food and Agriculture, 2017, 97(6):1945-1952.
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