氧化对大豆蛋白结构、乳液稳定性及消化特性的影响
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摘要
以大豆分离蛋白为原料,以脂质过氧化产物丙二醛(malondialdehyde,MDA)为氧化引发剂,逐级研究氧化对大豆蛋白结构、乳液稳定性及乳液消化特性的影响。结果发现:随着MDA浓度的升高,蛋白羰基及席夫碱含量明显升高而巯基含量显著降低。同时,MDA可促进蛋白聚集并诱导β-伴大豆球蛋白(7S)组分形成二硫键和非二硫键诱导的共价交联。进一步制备O/W型乳液,发现不同浓度MDA处理蛋白对乳液的形成影响较小,但可以显著改变界面蛋白组成。其中经中高浓度(2.5～10 mmol/L)MDA氧化后,更多7S组分以聚集状态参与界面组成。体外模拟胃肠道消化实验进一步表明,乳液消化主要在肠道进行,氧化诱导的蛋白交联/聚集可延缓或降低胆盐在界面的替代,进而减缓乳液消化并降低脂质释放率。
In this study, malondialdehyde(MDA), a natural lipid peroxidation product, was used as an initiator to investigate the influence of MDA-induced protein structural modifications on stability and digestibility of emulsion stabilized by soy protein isolate(SPI). Results obtained showed that the contents of protein carbonyl and Schiff base significantly increased along with increasing MDA concentration, accompanied by a noticeable decrease in protein sulfhydryl content. Meanwhile,MDA promoted protein aggregation through disulfide/non-disulfide covalent bonding, where β-conglycinin(7S) was mainly involved in the latter case. O/W emulsion was further prepared with oxidatively treated SPI. It was found that MDA-induced protein oxidation had less influence on emulsion formation, but they could significantly alter the protein composition of the interface. Herein, for SPI treated with MDA at concentrations of 2.5-10 mmol/L, more 7S fractions participated in interface formation in an aggregate state. Furthermore, results from in vitro simulated gastrointestinal digestion demonstrated that emulsion digestion occurred mainly in the intestinal tract, and oxidation-induced cross-linking/aggregation of proteins could delay or reduce the replacement of bile salts at the interface, which in turn slowed emulsion digestion and decreased lipid digestibility.
In this study, malondialdehyde(MDA), a natural lipid peroxidation product, was used as an initiator to investigate the influence of MDA-induced protein structural modifications on stability and digestibility of emulsion stabilized by soy protein isolate(SPI). Results obtained showed that the contents of protein carbonyl and Schiff base significantly increased along with increasing MDA concentration, accompanied by a noticeable decrease in protein sulfhydryl content. Meanwhile,MDA promoted protein aggregation through disulfide/non-disulfide covalent bonding, where β-conglycinin(7S) was mainly involved in the latter case. O/W emulsion was further prepared with oxidatively treated SPI. It was found that MDA-induced protein oxidation had less influence on emulsion formation, but they could significantly alter the protein composition of the interface. Herein, for SPI treated with MDA at concentrations of 2.5-10 mmol/L, more 7S fractions participated in interface formation in an aggregate state. Furthermore, results from in vitro simulated gastrointestinal digestion demonstrated that emulsion digestion occurred mainly in the intestinal tract, and oxidation-induced cross-linking/aggregation of proteins could delay or reduce the replacement of bile salts at the interface, which in turn slowed emulsion digestion and decreased lipid digestibility.
引文
[1]CHEN N N,ZHAO M M,SUN W Z,et al.Effect of oxidation on the emulsifying properties of soy protein isolate[J].Food Research International,2013,52(1):26-32.DOI:10.1016/j.foodres.2013.02.028.
[2]崔旭海,孔保华.蛋白质氧化及其对乳蛋白结构与功能性的影响[J].中国乳品工业,2008,36(1):44-47.DOI:10.3969/j.issn.1001-2230.2008.01.012.
[3]闫利国,唐善虎,王柳,等.冷冻贮藏过程中氧化诱导牦牛肉肌原纤维蛋白结构的变化[J].食品科学,2015,36(24):337-342.DOI:10.7506/spkx1002-6630-201524062.
[4]LIU Z,XIONG Y L,CHEN J.Protein oxidation enhances hydration but suppresses water-holding capacity in porcine longissimus muscle[J].Journal of Agricultural and Food Chemistry,2010,58(19):10697-10704.DOI:10.1021/jf102043k.
[5]XIA X F,KONG B H,XIONG Y L,et al.Decreased gelling and emulsifying properties of myofibrillar protein from repeatedly frozenthawed porcine longissimus muscle are due to protein denaturation and susceptibility to aggregation[J].Meat Science,2010,85(3):481-486.DOI:10.1016/j.meatsci.2010.02.019.
[6]DICKINSON E.Emulsion gels:the structuring of soft solids with protein-stabilized oil droplets[J].Food Hydrocolloids,2012,28(1):224-241.DOI:10.1016/j.foodhyd.2011.12.017.
[7]CHENG Q,DECKER E A,XIAO H,et al.Physical and chemical stability ofβ-carotene-enriched nanoemulsions:influence of pH,ionic strength,temperature,and emulsifier type[J].Food Chemistry,2012,132(3):1221-1229.DOI:10.1016/j.foodchem.2011.11.091.
[8]SINGH H,YE A,HORNE D.Structuring food emulsions in the gastrointestinal tract to modify lipid digestion[J].Progress in Lipid Research,2009,48(2):92-100.DOI:10.1016/j.plipres.2008.12.001.
[9]姚晓琳,聂珂,陈玉,等.复合界面对食品乳液中脂肪消化的影响[J].食品科学,2018,39(2):1-7.DOI:10.7506/spkx1002-6630-201802001.
[10]梁晗妮.豆类蛋白质的乳化性质及其结构.功能相关性研究[D].广州:华南理工大学,2014:36-48.
[11]SARKAR A,MURRAY B,HOLMES M,et al.In vitro digestion of Pickering emulsions stabilized by soft whey protein microgel particles:influence of thermal treatment[J].Soft Matter,2016,12(15):3558-3569.DOI:10.1039/c5sm02998h.
[12]ZHOU F B,SUN W Z,ZHAO M M.Controlled formation of emulsion gels stabilized by salted myofibrillar protein under malondialdehyde(MDA)-induced oxidative stress[J].Journal of Agricultural and Food Chemistry,2015,63(14):3766-3777.DOI:10.1021/jf505916f.
[13]WU W,WU X J,HUA Y F.Structural modification of soy protein by the lipid peroxidation product acrolein[J].Journal of the Science of Food&Agriculture,2009,89(8):1416-1423.DOI:10.1016/j.lwt.2009.05.006.
[14]MORZEL M,GATELLIER P,SAYD T,et al.Chemical oxidation decreases proteolytic susceptibility of skeletal muscle myofibrillar proteins[J].Meat Science,2006,73(3):536-543.DOI:10.1016/j.meatsci.2006.02.005.
[15]FLORES M,BARAT J M,ARISTORY M C,et al.Accelerated processing of dry-cured ham.Part 2.Influence of brine thawing/salting operation on proteolysis and sensory acceptability[J].Meat Science,2006,72(4):766.DOI:10.1016/j.meatsci.2005.10.008.
[16]TOKLE T,LESMES U,DECKER E A,et al.Impact of dietary fiber coatings on behavior of protein-stabilized lipid droplets under simulated gastrointestinal conditions[J].Food&Function,2012,3(1):58.DOI:10.1039/C1FO10129C.
[17]ZEEB B,LOPEZ-PENA C L,WEISS J,et al.Controlling lipid digestion using enzyme-induced crosslinking of biopolymer interfacial layers in multilayer emulsions[J].Food Hydrocolloids,2015,46:125-133.DOI:10.1016/j.foodhyd.2014.12.018.
[18]BURCHAM P C,KUHAN Y T.Introduction of carbonyl groups into proteins by the lipid peroxidation product,malondialdehyde[J].Biochemical and Biophysical Research Communications,1996,220(3):996-1001.DOI:10.1006/bbrc.1996.0521.
[19]ZHOU F B,ZHAO M M,SU G W,et al.Gelation of salted myofibrillar protein under malondialdehyde-induced oxidative stress[J].Food Hydrocolloids,2014,40(10):153-162.DOI:10.1016/j.foodhyd.2014.03.001.
[20]刘岩,赵冠里,赵谋明,等.花生湿热处理对其分离蛋白的结构和功能特性的影响[J].现代食品科技,2011,27(5):506-510.DOI:10.3969/j.issn.1673-9078.2011.05.005.
[21]王金梅,王梦萍,韦翠兰,等.大豆球蛋白的纤维聚集体行为及其稳定性研究[J].现代食品科技,2017,33(11):42-48.DOI:10.13982/j.mfst.1673-9078.2017.11.007.
[22]程翠林,石彦国,王振宇,等.大豆蛋白亚基组成对其功能特性的影响[J].食品科学,2006,27(3):70-74.DOI:10.3321/j.issn:1002-6630.2006.03.012.
[23]HUANG Y R,HUA Y F,QIU A Y.Soybean protein aggregation induced by lipoxygenase catalyzed linoleic acid oxidation[J].Food Research International,2006,39(2):240-249.DOI:10.1016/j.foodres.2005.07.012.
[24]WU W,ZHANG C,HUA Y F.Structural modification of soy protein by the lipid peroxidation product malondialdehyde[J].Journal of the Science of Food and Agriculture,2009,89(8):1416-1423.DOI:10.1002/jsfa.3606.
[25]CHEN N,ZHAO M J,SUN W,et al.Effect of oxidation on the emulsifying properties of soy protein isolate[J].Food Research International,2013,52(1):26-32.DOI:10.1016/j.foodres.2013.02.028.
[26]DAMODARAN S,RAZUMOVSKY L.Role of surface area-tovolume ratio in protein adsorption at the air-water interface[J].Surface Science,2008,602(1):307-315.DOI:10.1016/j.susc.2007.10.018.
[27]GOLDING M,WOOSTER T J.The influence of emulsion structure and stability on lipid digestion[J].Current Opinion in Colloid&Interface Science,2010,15(1):90-101.DOI:10.1016/j.cocis.2009.11.006.
[28]EYDOUX C,SPINELLI S,DAVIS T L,et al.Structure of human pancreatic lipase-related protein 2 with the lid in an open conformation[J].Biochemistry,2008,47(36):9553-9564.DOI:10.1021/bi8005576.
[29]李伟伟.高乳化性大豆蛋白的制备及其界面流变性质的研究[D].无锡:江南大学,2017:26-35.
[30]MUN S,DECKER E A,MCCLEMENTS D J.Influence of droplet characteristics on the formation of oil-in-water emulsions stabilized by surfactant-chitosan layers[J].Langmuir the ACS Journal of Surfaces&Colloids,2005,21(14):6228-6234.DOI:10.1021/la050502w.
[31]MALDONADO-VALDERRAMA J,WOODWARD N C,GUNNINGA P,et al.Interfacial characterization ofβ-lactoglobulin networks:displacement by bile salts[J].Langmuir,2008,24(13):6759-6767.DOI:10.1021/la800551u.
[32]BELLESI F A,RUIZ-HENESTROSA V M P,MALDONADO-VALDERRAMA J,et al.Comparative interfacial in vitro digestion of protein and polysaccharide oil/water films[J].Colloids and Surfaces B:Biointerfaces,2018,161:547-554.DOI:10.1016/j.colsurfb.2017.11.027.
[2]崔旭海,孔保华.蛋白质氧化及其对乳蛋白结构与功能性的影响[J].中国乳品工业,2008,36(1):44-47.DOI:10.3969/j.issn.1001-2230.2008.01.012.
[3]闫利国,唐善虎,王柳,等.冷冻贮藏过程中氧化诱导牦牛肉肌原纤维蛋白结构的变化[J].食品科学,2015,36(24):337-342.DOI:10.7506/spkx1002-6630-201524062.
[4]LIU Z,XIONG Y L,CHEN J.Protein oxidation enhances hydration but suppresses water-holding capacity in porcine longissimus muscle[J].Journal of Agricultural and Food Chemistry,2010,58(19):10697-10704.DOI:10.1021/jf102043k.
[5]XIA X F,KONG B H,XIONG Y L,et al.Decreased gelling and emulsifying properties of myofibrillar protein from repeatedly frozenthawed porcine longissimus muscle are due to protein denaturation and susceptibility to aggregation[J].Meat Science,2010,85(3):481-486.DOI:10.1016/j.meatsci.2010.02.019.
[6]DICKINSON E.Emulsion gels:the structuring of soft solids with protein-stabilized oil droplets[J].Food Hydrocolloids,2012,28(1):224-241.DOI:10.1016/j.foodhyd.2011.12.017.
[7]CHENG Q,DECKER E A,XIAO H,et al.Physical and chemical stability ofβ-carotene-enriched nanoemulsions:influence of pH,ionic strength,temperature,and emulsifier type[J].Food Chemistry,2012,132(3):1221-1229.DOI:10.1016/j.foodchem.2011.11.091.
[8]SINGH H,YE A,HORNE D.Structuring food emulsions in the gastrointestinal tract to modify lipid digestion[J].Progress in Lipid Research,2009,48(2):92-100.DOI:10.1016/j.plipres.2008.12.001.
[9]姚晓琳,聂珂,陈玉,等.复合界面对食品乳液中脂肪消化的影响[J].食品科学,2018,39(2):1-7.DOI:10.7506/spkx1002-6630-201802001.
[10]梁晗妮.豆类蛋白质的乳化性质及其结构.功能相关性研究[D].广州:华南理工大学,2014:36-48.
[11]SARKAR A,MURRAY B,HOLMES M,et al.In vitro digestion of Pickering emulsions stabilized by soft whey protein microgel particles:influence of thermal treatment[J].Soft Matter,2016,12(15):3558-3569.DOI:10.1039/c5sm02998h.
[12]ZHOU F B,SUN W Z,ZHAO M M.Controlled formation of emulsion gels stabilized by salted myofibrillar protein under malondialdehyde(MDA)-induced oxidative stress[J].Journal of Agricultural and Food Chemistry,2015,63(14):3766-3777.DOI:10.1021/jf505916f.
[13]WU W,WU X J,HUA Y F.Structural modification of soy protein by the lipid peroxidation product acrolein[J].Journal of the Science of Food&Agriculture,2009,89(8):1416-1423.DOI:10.1016/j.lwt.2009.05.006.
[14]MORZEL M,GATELLIER P,SAYD T,et al.Chemical oxidation decreases proteolytic susceptibility of skeletal muscle myofibrillar proteins[J].Meat Science,2006,73(3):536-543.DOI:10.1016/j.meatsci.2006.02.005.
[15]FLORES M,BARAT J M,ARISTORY M C,et al.Accelerated processing of dry-cured ham.Part 2.Influence of brine thawing/salting operation on proteolysis and sensory acceptability[J].Meat Science,2006,72(4):766.DOI:10.1016/j.meatsci.2005.10.008.
[16]TOKLE T,LESMES U,DECKER E A,et al.Impact of dietary fiber coatings on behavior of protein-stabilized lipid droplets under simulated gastrointestinal conditions[J].Food&Function,2012,3(1):58.DOI:10.1039/C1FO10129C.
[17]ZEEB B,LOPEZ-PENA C L,WEISS J,et al.Controlling lipid digestion using enzyme-induced crosslinking of biopolymer interfacial layers in multilayer emulsions[J].Food Hydrocolloids,2015,46:125-133.DOI:10.1016/j.foodhyd.2014.12.018.
[18]BURCHAM P C,KUHAN Y T.Introduction of carbonyl groups into proteins by the lipid peroxidation product,malondialdehyde[J].Biochemical and Biophysical Research Communications,1996,220(3):996-1001.DOI:10.1006/bbrc.1996.0521.
[19]ZHOU F B,ZHAO M M,SU G W,et al.Gelation of salted myofibrillar protein under malondialdehyde-induced oxidative stress[J].Food Hydrocolloids,2014,40(10):153-162.DOI:10.1016/j.foodhyd.2014.03.001.
[20]刘岩,赵冠里,赵谋明,等.花生湿热处理对其分离蛋白的结构和功能特性的影响[J].现代食品科技,2011,27(5):506-510.DOI:10.3969/j.issn.1673-9078.2011.05.005.
[21]王金梅,王梦萍,韦翠兰,等.大豆球蛋白的纤维聚集体行为及其稳定性研究[J].现代食品科技,2017,33(11):42-48.DOI:10.13982/j.mfst.1673-9078.2017.11.007.
[22]程翠林,石彦国,王振宇,等.大豆蛋白亚基组成对其功能特性的影响[J].食品科学,2006,27(3):70-74.DOI:10.3321/j.issn:1002-6630.2006.03.012.
[23]HUANG Y R,HUA Y F,QIU A Y.Soybean protein aggregation induced by lipoxygenase catalyzed linoleic acid oxidation[J].Food Research International,2006,39(2):240-249.DOI:10.1016/j.foodres.2005.07.012.
[24]WU W,ZHANG C,HUA Y F.Structural modification of soy protein by the lipid peroxidation product malondialdehyde[J].Journal of the Science of Food and Agriculture,2009,89(8):1416-1423.DOI:10.1002/jsfa.3606.
[25]CHEN N,ZHAO M J,SUN W,et al.Effect of oxidation on the emulsifying properties of soy protein isolate[J].Food Research International,2013,52(1):26-32.DOI:10.1016/j.foodres.2013.02.028.
[26]DAMODARAN S,RAZUMOVSKY L.Role of surface area-tovolume ratio in protein adsorption at the air-water interface[J].Surface Science,2008,602(1):307-315.DOI:10.1016/j.susc.2007.10.018.
[27]GOLDING M,WOOSTER T J.The influence of emulsion structure and stability on lipid digestion[J].Current Opinion in Colloid&Interface Science,2010,15(1):90-101.DOI:10.1016/j.cocis.2009.11.006.
[28]EYDOUX C,SPINELLI S,DAVIS T L,et al.Structure of human pancreatic lipase-related protein 2 with the lid in an open conformation[J].Biochemistry,2008,47(36):9553-9564.DOI:10.1021/bi8005576.
[29]李伟伟.高乳化性大豆蛋白的制备及其界面流变性质的研究[D].无锡:江南大学,2017:26-35.
[30]MUN S,DECKER E A,MCCLEMENTS D J.Influence of droplet characteristics on the formation of oil-in-water emulsions stabilized by surfactant-chitosan layers[J].Langmuir the ACS Journal of Surfaces&Colloids,2005,21(14):6228-6234.DOI:10.1021/la050502w.
[31]MALDONADO-VALDERRAMA J,WOODWARD N C,GUNNINGA P,et al.Interfacial characterization ofβ-lactoglobulin networks:displacement by bile salts[J].Langmuir,2008,24(13):6759-6767.DOI:10.1021/la800551u.
[32]BELLESI F A,RUIZ-HENESTROSA V M P,MALDONADO-VALDERRAMA J,et al.Comparative interfacial in vitro digestion of protein and polysaccharide oil/water films[J].Colloids and Surfaces B:Biointerfaces,2018,161:547-554.DOI:10.1016/j.colsurfb.2017.11.027.