大豆糖蛋白的抗氧化特性及其机理
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摘要
将大豆分离蛋白(SPI)与葡萄糖按质量比1∶2混合,通过美拉德反应生成大豆糖蛋白,测定不同反应温度(70,80,90℃)、不同反应时间(0,1,2,3,4,5,6h)条件的大豆糖蛋白的还原能力、羟自由基清除能力、DPPH自由基清除能力、游离氨基含量、褐变程度,紫外光谱扫描分析和聚丙烯酰氨凝胶电泳(SDS-PAGE)等指标,探讨大豆糖蛋白的抗氧化性及其作用机理。结果表明:随着反应时间的延长,大豆糖蛋白中游离氨基含量逐渐减少(P<0.05),反应褐变程度越大,抗氧化性越高,其中90℃6 h的大豆糖蛋白具有最高的还原能力,比SPI还原能力高5.6倍;90℃5 h的MRPs羟自由基清除能力与SPI相比提高了1.69倍,其DPPH自由基清除能力高达68.55%,是SPI的DPPH自由基清除能力的3.67倍。紫外扫描结果表明:MRPs的色氨酸吸收峰强度随着反应时间的延长逐渐增强,而80℃和90℃6 h MRPs色氨酸吸收峰有明显减弱趋势。SDS-PAGE电泳图谱表明,随着反应的进行,MRPs的α′和α亚基条带逐渐变浅,生成的大分子物质发生聚集。大豆蛋白葡萄糖反应生成的MRPs具有很强的抗氧化性,其抗氧化性与蛋白结构的变化密切相关。
Maillard reaction products(MRPs) were prepared from soy protein isolate(SPI) and glucose(MW=1 ∶ 2) by heating at 70 ℃, 80 ℃ or 90 ℃ for 0-6 h. The reducing ability, hydroxyl radical scavenging ability and DPPH radical scavenging ability were determined to investigate antioxidant of MRPs. Moreover, the degree of browning, UV spectroscopy scan analysis and SDS-PAGE were studied to explore antioxidant mechanism of MRPs. The results showed that free amino group content of MRPs were significantly decreased(P<0.05) as the reaction proceeded, the antioxidant activity of MRPs was enhanced with the increase degree browning. The oxidation resistance of MRPs were improved as the reaction temperature increased, it was proved that MRPs which is heated for 6 h at 90℃ has the maximum reducing power, which is 5.6 times higher than SPI. MRPs obtained for 5 h at 90 ℃ exhibited the highest hydroxyl radical scavenging(1.69-fold increase over SPI) and DPPH radical-scavenging activity(68.55% scavenging activity, a 3.67-fold increase over SPI). UV scan demonstrated that tryptophan absorption peak intensity of MRPs showed a trend of increase with increasing reaction time, whereas tryptophan absorption peak of MRPs heated for 6 h at 80 ℃ and 90 ℃ decreased. Meanwhile, SDS-PAGE illustrated that SPI and glucose formed high molecular weight conjugates, and the α' and α sub band becomes shallow gradually with the reaction of maillard. All in all, these results suggested that glycoprotein have antioxidant properties and there is a close relationship between oxidation resistance and the changes of protein structure.
Maillard reaction products(MRPs) were prepared from soy protein isolate(SPI) and glucose(MW=1 ∶ 2) by heating at 70 ℃, 80 ℃ or 90 ℃ for 0-6 h. The reducing ability, hydroxyl radical scavenging ability and DPPH radical scavenging ability were determined to investigate antioxidant of MRPs. Moreover, the degree of browning, UV spectroscopy scan analysis and SDS-PAGE were studied to explore antioxidant mechanism of MRPs. The results showed that free amino group content of MRPs were significantly decreased(P<0.05) as the reaction proceeded, the antioxidant activity of MRPs was enhanced with the increase degree browning. The oxidation resistance of MRPs were improved as the reaction temperature increased, it was proved that MRPs which is heated for 6 h at 90℃ has the maximum reducing power, which is 5.6 times higher than SPI. MRPs obtained for 5 h at 90 ℃ exhibited the highest hydroxyl radical scavenging(1.69-fold increase over SPI) and DPPH radical-scavenging activity(68.55% scavenging activity, a 3.67-fold increase over SPI). UV scan demonstrated that tryptophan absorption peak intensity of MRPs showed a trend of increase with increasing reaction time, whereas tryptophan absorption peak of MRPs heated for 6 h at 80 ℃ and 90 ℃ decreased. Meanwhile, SDS-PAGE illustrated that SPI and glucose formed high molecular weight conjugates, and the α' and α sub band becomes shallow gradually with the reaction of maillard. All in all, these results suggested that glycoprotein have antioxidant properties and there is a close relationship between oxidation resistance and the changes of protein structure.
引文
[1] ETHIER M, SABA J A, S PEARLLLAN M, et al.Application of the stroligoalgorithm for the automated structure assignment of complex N-linked glyeans from glyeoproteins using tandem mass spectrometry[J].Rapid Cornnun Mass Speetrom, 2003, 17(24):2713-2720.
[2] ZHOU F B, ZHAO M M, ZHAO H F, et al. Effects of oxidative modification on gel properties of isolated porcine myofibrillar protein by peroxyl radicals[J]. Meat Science, 2014, 96(4):1432-1439.
[3] GU X, CAMPBELL L J, EUSTON S R. Influence of sugars on the chaaeteristies of ueono-a-lactoneinduced soy protein isolate gels[J]. Food Hydrocolloids, 2009, 23(2):314-326.
[4]穆利霞.大豆蛋白-糖接枝改性及其结构与功能特性研究[D].广州:华南理工大学, 2012.
[5] AEHOUI A, BOYE J I, BELAN J I. Funetional and molecular properties of ealeium precipitated soy glycinin and the effect of glyeationwithe earrageenan[J]. Food Reseach Internatlonal, 2010, 43(4):1494-1504.
[6] MARTINS S L, JONGEN W M, VAN BOEKEL M A. A review of Maillard reaction in food and implications to kinetic modeling[J]. Trends in Food Science and Technology, 2001, 11(9/10):364-373.
[7] LIU Q, LU Y, HAN J C, CHEN Q, KONG B H.Structure-modification by moderate oxidation in hydroxyl radical-generating systems promote the emulsifying properties of soy protein isolate[J]. Food Structure, 2015, 6(10):21-28.
[8] ACOSTA-DOM魱NGUEZ L, HERN魣NDEZ-S魣NCHEZ H, GUTI魪RREZ-L魷PEZ G F, et al. Modification of the soy protein isolate surface at nanometric scale and its effect on physicochemical properties[J]. Journal of Food Engineering, 2016, 12(168):105-112.
[9] HU H, WU J H, EUNICE C Y, et al. Effects of ultrasound on structural and physical properties of soy protein isolate(SPI)dispersions[J]. Food Hydrocolloids, 2013, 30(2):647-655.
[10] LI W W, ZHAO H B, HE Z Y, et al. Modification of soy protein hydrolysates by Maillard reaction:Effects of carbohydrate chain length on structural and interfacial properties[J]. Colloids and Surfaces B:Biointerfaces, 2016, 138(1):70-77.
[11] LERTITTIKUL W, BENJAKUL S, TANAKA M.Characteristics and antioxidative activity of Maillard reaction products from a porcine plasma protein-glucose model system as influenced by pH[J]. Food Chemistry, 2007, 100(2):669-677
[12] DEEPAK A V, THIPPESWAMY G, SHIVAKAMESHWARI M M. Isolation and characterization of a 29-kDa glycoprotein with antifungal activity from bulbs of Urginea indica[J]. Biochemical and Biophysical Research Communications, 2003, 311(3):735-742.
[13]任国艳.海蜇口腕部糖蛋白理化性质及生物活性研究[D].青岛:中国海洋大学, 2008.
[14]王玲,朱秀清,李佳栋,许慧,郑环宇,吴海波.低温冷冻条件对大豆分离蛋白分散液表面疏水性及二硫键的影响[J].食品科学, 2014, 35(7):28-32.
[15] YOUNG V R. Soy protein in relation to human pro tein and amino acid nutrition[J]. Journal of the American Dietetic Association, 1991, 91(4):828-835.
[16]陈创前,蔡会武,邵纪生,等.大豆蛋白复合酶解法研究[J].安徽农业科学, 2011, 39(10):6070-6074.
[17]王中江,王晶,李杨,等.大豆蛋白改性修饰技术研究进展[J].中国食物与营养, 2014, 20(1):27-31.
[18] BRUDZYNSKI K, MIOTTO D. Honey melanoidins:Analysis of the compositions of the high molecular weight melanoidins exhibiting radical-scavenging activity[J]. Food Chemistry, 2011, 127(3):1023-1030.
[19] XIA X F, KONG B H, LIU J, DIAO X P, QIAN L. Influence of different thawing methods on physicochemical changes and protein oxidation of porcine longissimus muscle[J]. LWT-Food Science and Technology, 2012:46(1):280-286.
[20]任为聪,程建军,张智宇.不同改性方法对蛋白质溶解性的影响研究进展[J].中国粮油学报, 2011,26(8):124-128.
[21] LIU J, RU Q, DING Y. Glycation a promising method for food protein modification:physicochemical properties and structure, a review[J]. Food Research International, 2012, 49(1):170-183.
[22] PARRELLA A, CATERINO E, CANGIANO M, ET A L. Antioxidant properties of different milk fermented with lactic acid bacteria and yeast[J]. International Journal of Food Science and Technology,2012, 2(47):2493-2502.
[23] TIAN S, CHEN J I E, SMALL D M. Enhancement of solubility and emulsifying properties of soy protein isolates by glucose conjugation[J]. Journal of Food Processing and Preservation, 2011, 35(1):80-95.
[24] SHAND P J, YA H, PIETRASIK Z. Physicochemical and textural properties of heat-induced pea protein isolate gels[J]. Food Chemistry, 2007, 102(4):1119-1130.
[2] ZHOU F B, ZHAO M M, ZHAO H F, et al. Effects of oxidative modification on gel properties of isolated porcine myofibrillar protein by peroxyl radicals[J]. Meat Science, 2014, 96(4):1432-1439.
[3] GU X, CAMPBELL L J, EUSTON S R. Influence of sugars on the chaaeteristies of ueono-a-lactoneinduced soy protein isolate gels[J]. Food Hydrocolloids, 2009, 23(2):314-326.
[4]穆利霞.大豆蛋白-糖接枝改性及其结构与功能特性研究[D].广州:华南理工大学, 2012.
[5] AEHOUI A, BOYE J I, BELAN J I. Funetional and molecular properties of ealeium precipitated soy glycinin and the effect of glyeationwithe earrageenan[J]. Food Reseach Internatlonal, 2010, 43(4):1494-1504.
[6] MARTINS S L, JONGEN W M, VAN BOEKEL M A. A review of Maillard reaction in food and implications to kinetic modeling[J]. Trends in Food Science and Technology, 2001, 11(9/10):364-373.
[7] LIU Q, LU Y, HAN J C, CHEN Q, KONG B H.Structure-modification by moderate oxidation in hydroxyl radical-generating systems promote the emulsifying properties of soy protein isolate[J]. Food Structure, 2015, 6(10):21-28.
[8] ACOSTA-DOM魱NGUEZ L, HERN魣NDEZ-S魣NCHEZ H, GUTI魪RREZ-L魷PEZ G F, et al. Modification of the soy protein isolate surface at nanometric scale and its effect on physicochemical properties[J]. Journal of Food Engineering, 2016, 12(168):105-112.
[9] HU H, WU J H, EUNICE C Y, et al. Effects of ultrasound on structural and physical properties of soy protein isolate(SPI)dispersions[J]. Food Hydrocolloids, 2013, 30(2):647-655.
[10] LI W W, ZHAO H B, HE Z Y, et al. Modification of soy protein hydrolysates by Maillard reaction:Effects of carbohydrate chain length on structural and interfacial properties[J]. Colloids and Surfaces B:Biointerfaces, 2016, 138(1):70-77.
[11] LERTITTIKUL W, BENJAKUL S, TANAKA M.Characteristics and antioxidative activity of Maillard reaction products from a porcine plasma protein-glucose model system as influenced by pH[J]. Food Chemistry, 2007, 100(2):669-677
[12] DEEPAK A V, THIPPESWAMY G, SHIVAKAMESHWARI M M. Isolation and characterization of a 29-kDa glycoprotein with antifungal activity from bulbs of Urginea indica[J]. Biochemical and Biophysical Research Communications, 2003, 311(3):735-742.
[13]任国艳.海蜇口腕部糖蛋白理化性质及生物活性研究[D].青岛:中国海洋大学, 2008.
[14]王玲,朱秀清,李佳栋,许慧,郑环宇,吴海波.低温冷冻条件对大豆分离蛋白分散液表面疏水性及二硫键的影响[J].食品科学, 2014, 35(7):28-32.
[15] YOUNG V R. Soy protein in relation to human pro tein and amino acid nutrition[J]. Journal of the American Dietetic Association, 1991, 91(4):828-835.
[16]陈创前,蔡会武,邵纪生,等.大豆蛋白复合酶解法研究[J].安徽农业科学, 2011, 39(10):6070-6074.
[17]王中江,王晶,李杨,等.大豆蛋白改性修饰技术研究进展[J].中国食物与营养, 2014, 20(1):27-31.
[18] BRUDZYNSKI K, MIOTTO D. Honey melanoidins:Analysis of the compositions of the high molecular weight melanoidins exhibiting radical-scavenging activity[J]. Food Chemistry, 2011, 127(3):1023-1030.
[19] XIA X F, KONG B H, LIU J, DIAO X P, QIAN L. Influence of different thawing methods on physicochemical changes and protein oxidation of porcine longissimus muscle[J]. LWT-Food Science and Technology, 2012:46(1):280-286.
[20]任为聪,程建军,张智宇.不同改性方法对蛋白质溶解性的影响研究进展[J].中国粮油学报, 2011,26(8):124-128.
[21] LIU J, RU Q, DING Y. Glycation a promising method for food protein modification:physicochemical properties and structure, a review[J]. Food Research International, 2012, 49(1):170-183.
[22] PARRELLA A, CATERINO E, CANGIANO M, ET A L. Antioxidant properties of different milk fermented with lactic acid bacteria and yeast[J]. International Journal of Food Science and Technology,2012, 2(47):2493-2502.
[23] TIAN S, CHEN J I E, SMALL D M. Enhancement of solubility and emulsifying properties of soy protein isolates by glucose conjugation[J]. Journal of Food Processing and Preservation, 2011, 35(1):80-95.
[24] SHAND P J, YA H, PIETRASIK Z. Physicochemical and textural properties of heat-induced pea protein isolate gels[J]. Food Chemistry, 2007, 102(4):1119-1130.
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