核桃多酚与蛋白质的作用方式及其抗氧化活性
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摘要
以核桃为研究对象,将核桃碱提蛋白和酸提蛋白经溶剂作用后超滤,通过破坏核桃多酚-蛋白复合体间的作用力得到不同结合态多酚,测定多酚含量及其抗氧化活性,以期探究核桃酚类物质与核桃蛋白化学键结合机制。研究结果表明,在pH3.5和pH11条件下,游离态多酚含量分别为6.46、6.27 mg/g,占总酚含量的25.15%、29.77%,非共价结合态多酚含量分别为11.31、7.74 mg/g,占总酚含量的44.02%、36.75%,共价结合态含量7.92、7.05 mg/g,占总酚含量的30.83%、33.48%。由此可知,非共价结合为核桃多酚与核桃蛋白质的主要结合方式。在pH3.5和pH11条件下,游离态多酚对DPPH自由基的清除率分别为82.34%、43.68%,Fe~(2+)螯合率分别为71.88%、49.96%,还原力吸光度值分别为1.157、0.857,脂质抗过氧化能力分别为47.71%、32.69%。在5种不同结合态多酚中,游离态多酚的抗氧化活性显著高于其他结合态多酚(p<0.05)。
The mechanism of chemical bonding between walnut phenolic compounds and walnut protein of the walnut were explored in this paper. The walnut protein solution was first treated by solvent under alkaline and acidic conditions,and then ultrafiltration was carried out,and different bound polyphenols were obtained by destroying the interaction force between walnut protein-polyphenol compounds. The content and antioxidant activity of polyphenols were determined. The results showed that the contents of free polyphenols were 6.46,6.27 mg/g respectively,accounting for 25.15% and 29.77% of the total phenols,the contents of non-covalent polyphenols were 11.31,7.74 mg/g respectively,accounting for 44.02% and 36.75% of the total phenols,and the contents of covalent polyphenols were 7.92,7.05 mg/g respectively,accounting for 30.83% and 33.48% of the total phenols.It was known that non covalent binding was the main form of interaction between polyphenols and protein complexes. Under pH3.5 and pH11 conditions,the scavenging rates of free polyphenols on DPPH free radicals were 82.34% and 43.68%,Fe~(2+) chelating ability were 71.88% and 49.96%,reducing power absorbance value were 1.157 and 0.857,inhibition of lipid peroxidation ability were 47.71% and 32.69%,respectively. Among the five different bound polyphenols,the antioxidant capacity of free polyphenols was significantly higher than that of other bound polyphenols(p<0.05).
The mechanism of chemical bonding between walnut phenolic compounds and walnut protein of the walnut were explored in this paper. The walnut protein solution was first treated by solvent under alkaline and acidic conditions,and then ultrafiltration was carried out,and different bound polyphenols were obtained by destroying the interaction force between walnut protein-polyphenol compounds. The content and antioxidant activity of polyphenols were determined. The results showed that the contents of free polyphenols were 6.46,6.27 mg/g respectively,accounting for 25.15% and 29.77% of the total phenols,the contents of non-covalent polyphenols were 11.31,7.74 mg/g respectively,accounting for 44.02% and 36.75% of the total phenols,and the contents of covalent polyphenols were 7.92,7.05 mg/g respectively,accounting for 30.83% and 33.48% of the total phenols.It was known that non covalent binding was the main form of interaction between polyphenols and protein complexes. Under pH3.5 and pH11 conditions,the scavenging rates of free polyphenols on DPPH free radicals were 82.34% and 43.68%,Fe~(2+) chelating ability were 71.88% and 49.96%,reducing power absorbance value were 1.157 and 0.857,inhibition of lipid peroxidation ability were 47.71% and 32.69%,respectively. Among the five different bound polyphenols,the antioxidant capacity of free polyphenols was significantly higher than that of other bound polyphenols(p<0.05).
引文
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[21]代沙.紫苏叶抗氧化物质提取、含量测定及抗氧化活性研究[D].雅安:四川农业大学,2013.
[22]王廷华,张云辉,邹晓莉.蛋白质理论与技术[M].北京:科学出版社,2013:28.
[23]贾娜,刘丹,谢振峰.植物多酚与食品蛋白质的相互作用[J].食品与发酵工业,2016,42(7):277-282.
[24]Subramanian V,MadhanB,Fathima N,et al.Hydration in matrix stabilization of collagen fibers:Relevance to mechanistic insight into tanning[J].Journal-American Leather Chemists Association,2003,98(7):263-272.
[25]Haslam E.Natural polyphenols(vegetable tannins)as drugs:possible modes of action[J].Journal of Natural Products,1996,59(2):205-215.
[26]Richard T,Vitrac X,Merillon J M,et al.Role of peptide primary sequence in polyphenol-protein recognition:An example with neurotensin[J].Biochimica et Biophysica Acta,2005,1726(3):238-243.
[27]Abugoch L E,Martínez E N,Aňón M C.Influence of the extracting solvent upon the structural properties of Amaranth(Amaranthus hypochondriacus)glutelin[J].Journal of Agricultural & Food Chemistry,2003,51(14):4060-4065.
[28]黄惠华,王志,陈建新.多酚-蛋白质络合反应的影响因素研究[J].食品科学,2003,24(2):22-25.
[29]Mateus V D F.Structural features of procyanidin interactions with salivary proteins[J].J Agric Food Chem,2001,49(2):940-945.
[30]Kroll N G,Rawel H M,Rohn S.Reactions of plant phenolics with food proteins and enzymes under special consideration of covalent bonds[J].Food Science and Technology Research,2003,9(3):205-218.
[31]Alu’datt M H,Rababah T,Ereifej K,et al.Phenolic-protein interactions in oilseed protein isolates[J].Food Research International,2013,52(1):178-184.
[32]Ross K A,Beta T S,Arntfield S D.A comparative study on the phenolic acids identified and quantified in dry beans using HPLC as affected by different extraction and hydrolysis methods[J].Food Chemistry,2009,113(1):336-344.
[33]Lloyd B J,Siebenmorgen T J,Beers K W.Effects of commercial processing on antioxidants in rice bran 1[J].Cereal Chemistry,2007,77(5):551-555.
[34]章林,冷雪娇,黄明.鼠尾草提取物多酚含量与抗氧化活性的研究[J].食品工业科技,2012,33(19):143-146.
[35]王会,郭立,谢文磊.抗氧化剂抗氧化活性的测定方法(一)[J].食品与发酵工业,2006,32(4):98-102.
[2]陆俊,赵安琪,成策,等.核桃营养成分与生理活性及开发利用[J].食品与机械,2014(6):238-242.
[3]Szetao K W C,Sathe S K.Walnuts(Juglans regia L):proximate composition,protein solubility,protein amino acid composition and protein in vitro digestibility[J].Journal of the Science of Food & Agriculture,2000,80(9):1393-1401.
[4]郝艳宾.核桃仁中的多酚及其抗氧化活性的初步研究[A].中国园艺学会干果分会.第四届全国干果生产、科研进展研讨会论文集[C].中国园艺学会干果分会:中国园艺学会,2005:5.
[5]Yang J,Liu R H,Halim L.Antioxidant and antiproliferative activities of common edible nut seeds[J].LWT-Food Science and Technology,2009,42(1):1-8.
[6]刘畅,周家春.植物多酚抗氧化性研究[J].粮食与油脂,2011(2):43-46.
[7]Riceevans C A.Antioxidant properties of phenolic compounds[J].Trends in Plant Science,1997,2(4):152-159.
[8]刘夫国,马翠翠,王迪,等.蛋白质与多酚相互作用研究进展[J].食品与发酵工业,2016,42(2):282-288.
[9]Le Bourvellec C,Renard C M G C.Interactions between polyphenols and macromolecules:Quantification methods and mechanisms[J].Critical Reviews in Food Science & Nutrition,2012,52(3):213-248.
[10]李杨,孙红波,董济萱,等.花青素与大豆分离蛋白非共价/共价作用对其界面功能性质的影响[J].食品科学,2018,39(6):32-38.
[11]5种籼稻品种谷壳中游离态和结合态酚类物质含量及其抗氧化活性比较[J].中国农业科学,2012,45(6):1150-1158.
[12]Xu L,Diosady L L.Interactions between canola proteins and phenolic compounds in aqueous media[J].Food Research International,2000,33(9):725-731.
[13]Nardini M,Ghiselli A.Determination of free and bound phenolic acids in beer[J].Food Chemistry,2004,84(1):137-143.
[14]赵扬帆,郑宝东.植物多酚类物质及其功能学研究进展[J].福建轻纺,2006(11):107-110.
[15]毛晓英.核桃蛋白质的结构表征及其制品的改性研究[D].无锡:江南大学,2012.
[16]Shiwei S,Ruican W,Shuntang G,et al.Walnut phenolic compounds:Binding with proteins and antioxidant activities[J].Transactions of the Chinese Society of Agricultural Engineering,2016,32(22):309-314.
[17]蒲成伟,阚欢,刘云.核桃仁皮不同极性提取物的体外抗氧化活性分析[J].食品工业科技,2018(2):65-69.
[18]Zhang J,Zhang H,Wang L,et al.Antioxidant activities of the rice endosperm protein hydrolysate:Identification of the active peptide[J].European Food Research and Technology,2009,229(4):709-719.
[19]Karama M.Chelation of Cu(II),Zn(II),and Fe(II)by tannin constituents of selected edible nuts[J].International Journal of Molecular Sciences,2009,10(12):5485-5497.
[20]李笑笑.核桃内种皮多酚的提取及核桃油与核桃蛋白粉的稳定性研究[D].无锡:江南大学,2017.
[21]代沙.紫苏叶抗氧化物质提取、含量测定及抗氧化活性研究[D].雅安:四川农业大学,2013.
[22]王廷华,张云辉,邹晓莉.蛋白质理论与技术[M].北京:科学出版社,2013:28.
[23]贾娜,刘丹,谢振峰.植物多酚与食品蛋白质的相互作用[J].食品与发酵工业,2016,42(7):277-282.
[24]Subramanian V,MadhanB,Fathima N,et al.Hydration in matrix stabilization of collagen fibers:Relevance to mechanistic insight into tanning[J].Journal-American Leather Chemists Association,2003,98(7):263-272.
[25]Haslam E.Natural polyphenols(vegetable tannins)as drugs:possible modes of action[J].Journal of Natural Products,1996,59(2):205-215.
[26]Richard T,Vitrac X,Merillon J M,et al.Role of peptide primary sequence in polyphenol-protein recognition:An example with neurotensin[J].Biochimica et Biophysica Acta,2005,1726(3):238-243.
[27]Abugoch L E,Martínez E N,Aňón M C.Influence of the extracting solvent upon the structural properties of Amaranth(Amaranthus hypochondriacus)glutelin[J].Journal of Agricultural & Food Chemistry,2003,51(14):4060-4065.
[28]黄惠华,王志,陈建新.多酚-蛋白质络合反应的影响因素研究[J].食品科学,2003,24(2):22-25.
[29]Mateus V D F.Structural features of procyanidin interactions with salivary proteins[J].J Agric Food Chem,2001,49(2):940-945.
[30]Kroll N G,Rawel H M,Rohn S.Reactions of plant phenolics with food proteins and enzymes under special consideration of covalent bonds[J].Food Science and Technology Research,2003,9(3):205-218.
[31]Alu’datt M H,Rababah T,Ereifej K,et al.Phenolic-protein interactions in oilseed protein isolates[J].Food Research International,2013,52(1):178-184.
[32]Ross K A,Beta T S,Arntfield S D.A comparative study on the phenolic acids identified and quantified in dry beans using HPLC as affected by different extraction and hydrolysis methods[J].Food Chemistry,2009,113(1):336-344.
[33]Lloyd B J,Siebenmorgen T J,Beers K W.Effects of commercial processing on antioxidants in rice bran 1[J].Cereal Chemistry,2007,77(5):551-555.
[34]章林,冷雪娇,黄明.鼠尾草提取物多酚含量与抗氧化活性的研究[J].食品工业科技,2012,33(19):143-146.
[35]王会,郭立,谢文磊.抗氧化剂抗氧化活性的测定方法(一)[J].食品与发酵工业,2006,32(4):98-102.