Abstract
［Objective］ To explore the interaction mechanism between diethyl phthalate (DEP ) and ovalbumin (OVA ) using multispectral analysis and molecular docking techniques.［Methods］ The Stern -Volmer equation is applied to study the fluorescence quenching mechanism of OVA by DEP,as well as to determine the binding constants and the number of binding sites between them.Thermodynamic parameters are used to identify the primary types of interaction forces.Based on Förster's non -radiative energy transfer theory,the energy transfer efficiency and binding distance between DEP and OVA are calculated.Molecular docking simulations are used to investigate the interaction forces between DEP and OVA.［Results］］ DEP can statically quench the intrinsic fluorescence of OVA.At temperatures ranging from 25 to 45 ℃,the binding constant (K) decreases from 2.631×108 L/mol to 1.072×108 L/mol,with 1~2 major binding sites identified between DEP and OVA.Thermodynamic analysis suggests that the interaction occurs spontaneously through electrostatic forces.Fluorescence resonance energy transfer parameters (J=1.316×10-13 cm3·L/mol,R0=3.665 nm,E=0.735,r=3.091 nm) confirms the strong binding.UV-Vis spectroscopy shows that DEP and OVA form a ground -state complex.Molecular docking simulations demonstrate 引用格式：付晨，黄群.光谱法联合分子对接研究邻苯二甲酸二乙酯与卵白蛋白的相互作用 ［J］.食品与机械，2025，41（7）：1-8.C itation:FU Chen,HUANG Qun.Investigation of the interaction between diethyl phthalate and ovalbumin via spectroscopic techniques and molecular docking [J].Food & Machinery,2025,41(7):1-8.that DEP binds to OVA through hydrogen bonds,van der Waals forces,and hydrophobic interactions,resulting in a stable complex structure.［Conclusion］ DEP can statically quench the intrinsic fluorescence of OVA.The two molecules spontaneously form a stable ground -state complex with 1~2 binding sites,a binding distance of 3.091 nm,and an energy transfer efficiency of 0.735.The stability of the complex structure is primarily maintained by hydrogen bonds,van der Waals forces,and hydrophobic interactions.
Publication Date
7-11-2025
First Page
1
Last Page
8
DOI
10.13652/j.spjx.1003.5788.2025.80427
Recommended Citation
Chen, FU and Qun, HUANG
(2025)
"Investigation of the interaction between diethyl phthalate and ovalbumin via spectroscopic techniques and molecular docking,"
Food and Machinery: Vol. 41:
Iss.
7, Article 1.
DOI: 10.13652/j.spjx.1003.5788.2025.80427
Available at:
https://www.ifoodmm.cn/journal/vol41/iss7/1
References
[1] 兰旭凯.邻苯二甲酸酯和微塑料对厚壳贻贝生殖毒性的研究[D].上海:上海海洋大学,2024:4-5.LAN X K.Study on the toxicity of phthalic acid ester and microplastics to Mytilus coruscus [D].Shanghai:Shanghai Ocean University,2024:4-5.
[2] MONDAL S,BASU S,GHOSH S,et al.Diethyl phthalate,a plasticizer,induces adipocyte inflammation and apoptosis in mice after long-term dietary administration [J].Journal of Biochemical and Molecular Toxicology,2024,38(1):e23561.
[3] MEI Y F,YANG W T,PENG S C,et al.Contamination levels and ecological risk assessment of phthalate esters (PAEs ) in the aquatic environment of Chaohu Lake [J].Environmental Science and Pollution Research International,2024,31(33):45 553-45 563.
[4] DU H,HU R W,ZHAO H M,et al.Mechanistic insight into esterase-catalyzed hydrolysis of phthalate esters (PAEs ) based on integrated multi-spectroscopic analyses and docking simulation [J].Journal of Hazardous Materials,2021,408:124901.
[5] CHENG J L,DU H,ZHOU M S,et al.Substrate-enzyme interactions and catalytic mechanism in a novel family VI esterase with dibutyl phthalate-hydrolyzing activity [J].Environment International,2023,178:108054.
[6] GUAN R,WANG Y Y,LIU H B,et al.Investigation on the interaction between catalase and typical phthalates with different side chain lengths [J].Spectochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2025,336:126031.[7] CEN C N,CHEN J,WANG W Q,et al.Exploring the interaction mechanism of dietary protein ovalbumin and folic acid:a combination research of molecular simulation technology and multispectroscopy [J].Food Chemistry,2022,385:132536.
[8] FU X Z,BELWAL T,HE Y H,et al.Interaction and binding mechanism of cyanidin- 3-O-glucoside to ovalbumin in varying pH conditions:a spectroscopic and molecular docking study [J].Food Chemistry,2020,320:126616.
[9] LIU Y J,MA L Y,ZHANG Q,et al.Construction of fatty acid-ovalbumin binary complexes to improve the water dispersibility,thermal/digestive stability and bioaccessibility of lutein:a comparative study of different fatty acids [J].International Journal of Biological Macromolecules,2024,273:133010.
[10] LIU T T,ZHAO Y,WU N,et al.Egg white protein-based delivery system for bioactive substances:a review [J].Critical Reviews in Food Science and Nutrition,2024,64(3):617-637.
[11] NUDRAT S,GIRI P,NONGBRI D L,et al.Analysis of the binding mechanism of bioactive coumarins with ovalbumin:further investigation into the inhibitory effects toward protein fibrillation [J].ACS Food Science & Technology,2023,3(5):866-880.
[12] CHEN Q,DONG L Z,LI Y,et al.Research advance of non-thermal processing technologies on ovalbumin properties:the gelation,foaming,emulsification,allergenicity,immunoregulation and its delivery system application [J].Critical Reviews in Food Science and Nutrition,2024,64(20):7 045-7 066.
[13] 梁鑫富,董庆亮.光谱法结合分子对接技术研究柠檬苦素与牛血清白蛋白的相互作用 [J].食品工业科技,2024,45(21):37-44.LIANG X F,DONG Q L.Spectroscopic method combined with molecular docking technique to study the interaction of limonin with bovine serum albumin [J].Science and Technology of Food Industry,2024,45(21):37-44.
[14] ZHANG Q Z,HUANG Z J,WANG Y,et al.Chinese bayberry (Myrica rubra ) phenolics mitigated protein glycoxidation and formation of advanced glycation end-products:a mechanistic investigation [J].Food Chemistry,2021,361:130102.
[15] LEITE MILIÃO G,DE SOUZA SOARES L,BALBINO D F,et al.pH influence on the mechanisms of interaction between chitosan and ovalbumin:a multi-spectroscopic approach [J].Food Hydrocolloids,2022,123:107137.
[16] HABIBI N,CHRISTAU S,OCHYL L J,et al.Engineered ovalbumin nanoparticles for cancer immunotherapy [J].Advanced Therapeutics,2020,3(10):2000100.
[17] 马兰,吴雨婷,周俊锹,等.多光谱法结合分子模拟探究邻苯二甲酸酯对胃蛋白酶的作用机理 [J].分析试验室,2024,43(10):1 377-1 384.MA L,WU Y T,ZHOU J Q,et al.Evaluation of the binding mechanisms of phthalates to pepsin based on multi-spectral analysis combined with computer simulation [J].Chinese Journal of Analytical Laboratory,2024,43(10):1 377-1 384.
[18] ZHANG Q X,CHEN Y Y,LIU W Y,et al.Effects of tweens on the structure,interfacial characteristics,and emulsifying and foaming properties of ovalbumin [J].Food Research International,2025,203:115824.
[19] ZHANG P,HUANG X Y,FU C,et al.Binding mechanism of Monascus pigment and ovalbumin:spectral analysis,molecular docking and molecular dynamics simulation [J].Food Science of Animal Products,2023,1(4):9240038.
[20] MA Y Y,ZHANG D D,LIU M Q,et al.Identification of antioxidant peptides derived from tilapia (Oreochromis niloticus ) skin and their mechanism of action by molecular docking [J].Foods,2022,11(17):2 576.
[21] CHENG W W,MA J L,WANG S J,et al.Interaction mechanism between resveratrol and ovalbumin based on fluorescence spectroscopy and molecular dynamic simulation[J].LWT-Food Science ang Technology,2021,146:111455.
[22] CHEN L,ZHU M,HU X,et al.Exploring the binding mechanism of ferulic acid and ovalbumin:insights from spectroscopy,molecular docking and dynamics simulation [J].Journal of the Science of Food and Agriculture,2022,102(9):3 835-3 846.
[23] KAFFASH M,TOLOU-SHIKHZADEH-YAZDI S,SOLEIMANI S,et al.Spectroscopy and molecular simulation on the interaction of Nano-Kaempferol prepared by oil-in-water with two carrier proteins:an investigation of protein-protein interaction [J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2024,309:123815.
[24] SAMUEL H S,NWEKW-MARAIZU U,ETIM E E.Understanding intermolecular and intramolecular hydrogen bonds:spectroscopic and computational approaches [J].Journal of Chemical Reviews,2023,5(4):439-465.
[25] GUO J Y,GAN C F,CHENG B,et al.Exploration of binding mechanism of apigenin to pepsin:spectroscopic analysis,molecular docking,enzyme activity and antioxidant assays [J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2023,290:122281.
[26] EFTINK M R,GHIRON C A.Fluorescence quenching studies with proteins [J].Analytical Biochemistry,1981,114(2):199-227.
[27] 杨彩英.热诱导卵白蛋白与白藜芦醇的作用机理及其复合物的致敏性评价 [D].南昌:南昌大学,2024:16.YANG C Y.Interaction mechanism of heat-stressed ovalbumin with resveratrol and the allergenicity evaluation of the complex[D].Nanchang:Nanchang University,2024:16.
[28] YU X,CAI X H,LI S,et al.Studies on the interactions of theaflavin- 3,3’-digallate with bovine serum albumin:multi-spectroscopic analysis and molecular docking [J].Food Chemistry,2022,366:130422.
[29] XIE H J,LI H,LIU L,et al.How the ovalbumin modulates the conformation of zein through protein-protein interactions [J].Food Hydrocolloids,2025,159:110696.
[30] CHEN F P,ZHOU L K,ZHOU B J,et al.Elucidation on the interaction between transferrin and ascorbic acid:a study based on spectroscopic analysis,molecular docking technology,and antioxidant evaluation [J].Journal of Molecular Liquids,2022,360:119413.
[31] ROSS P D,SUBRAMANIAN S.Thermodynamics of protein association reactions:forces contributing to stability [J].Biochemistry,1981,20(11):3 096-3 102.
[32] 侯小伟,张澍檀,严雪伟,等.液液萃取—气相色谱—质谱法测定水中邻苯二甲酸酯类化合物 [J].河南化工,2025,42(4):57-59.HOU X W,ZHANG S T,YAN X W,et al.Determination of phthalates in water by liquid extraction gas chromatography mass spectrometry [J].Henan Chemical Industry,2025,42(4):57-59.
[33] LYNDEM S,HAZARIKA U,ATHUL P,et al.A comprehensive in vitro exploration into the interaction mechanism of coumarin derivatives with bovine hemoglobin:spectroscopic and computational methods [J].Journal of Photochemistry and Photobiology A:Chemistry,2023,436:114425.
[34] ZHANG Y Y,LU Y C,YANG Y,et al.Comparison of non-covalent binding interactions between three whey proteins and chlorogenic acid:spectroscopic analysis and molecular docking[J].Food Bioscience,2021,41:101035.
[35] GAO Y,ZHOU Y,ZHOU X J,et al.Theoretical and observed mechanisms of diesel exhaust particles in promoting asthma[J].Chinese Medical Journal,2023,136(19):2 374-2 376.
[36] LIU Y J,CAI Y X,YING D Y,et al.Ovalbumin as a carrier to significantly enhance the aqueous solubility and photostability of curcumin:interaction and binding mechanism study [J].International Journal of Biological Macromolecules,2018,116:893-900.