过氧自由基氧化对米糠蛋白结构和功能性质的影响
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摘要
采用不同浓度的2,2’-盐酸脒基丙烷(2,2’-azobis(2-amidinopropane),AAPH)有氧热分解产生的过氧自由基氧化米糠蛋白,研究过氧自由基氧化对米糠蛋白结构和功能性质的影响。结果显示:随着AAPH浓度的增加,米糠蛋白羰基、二硫键和二酪氨酸含量分别从2.0 nmol/mg、12.04 nmol/mg和84.54增加到7.09 nmol/mg、14.69 nmol/mg和127.1,游离巯基含量从23.97 nmol/mg下降到15.29 nmol/mg。过氧自由基氧化导致米糠蛋白α-螺旋和无规卷曲相对含量下降,β-折叠相对含量增加,氨基酸残基侧链含量先增加后略微下降;同时使得米糠蛋白表面疏水性和内源荧光强度下降,最大荧光峰位蓝移。过氧自由基氧化对蛋白质亚基结构影响较小,可同时使得米糠蛋白多肽链聚集和主肽链部分断裂形成多肽。随着AAPH浓度的增加,米糠蛋白溶解性逐渐降低,持水性、持油性、起泡能力、泡沫稳定性、乳化性和乳化稳定性先上升后下降。
Effects of oxidative modification by peroxyl radicals derived from oxidative thermal decomposition of2,2'-azobis(2-amidinopropane)(AAPH) at different concentrations on the structural and functional properties of rice bran protein were investigated. The results indicated that as AAPH concentration increased, protein carbonyl, disulfide,and dityrosine contents of rice bran protein increased from 2.0 nmol/mg, 12.04 nmol/mg, and 84.54 to 7.09 nmol/mg,14.69 nmol/mg, and 127.1, respectively, while free sulfhydryl content decreased from 23.97 to 15.29 nmol/mg. Oxidative modification by peroxyl radicals resulted in a decrease in α-helix and random coil content and an increase in β-sheet content.Besides, the content of amino acid side chains firstly increased, and then slightly decreased after this treatment, which also led to a decreased in surface hydrophobicity and intrinsic fluorescence intensity accompanied by a blue shift of the maximum emission wavelength. Nevertheless, oxidative modification by peroxyl radicals had no significant impact on the subunit structure of rice bran protein, but caused polypeptide chain aggregation of rice bran protein accompanied by partial rupture of the major polypeptide chain to form peptides. As AAPH concentration increased, rice bran protein solubility gradually decreased; however, water-holding capacity, oil-holding capacity, foaming activity, foaming stability, emulsifying activity,and emulsiong stability firstly increased and then decreased.
Effects of oxidative modification by peroxyl radicals derived from oxidative thermal decomposition of2,2'-azobis(2-amidinopropane)(AAPH) at different concentrations on the structural and functional properties of rice bran protein were investigated. The results indicated that as AAPH concentration increased, protein carbonyl, disulfide,and dityrosine contents of rice bran protein increased from 2.0 nmol/mg, 12.04 nmol/mg, and 84.54 to 7.09 nmol/mg,14.69 nmol/mg, and 127.1, respectively, while free sulfhydryl content decreased from 23.97 to 15.29 nmol/mg. Oxidative modification by peroxyl radicals resulted in a decrease in α-helix and random coil content and an increase in β-sheet content.Besides, the content of amino acid side chains firstly increased, and then slightly decreased after this treatment, which also led to a decreased in surface hydrophobicity and intrinsic fluorescence intensity accompanied by a blue shift of the maximum emission wavelength. Nevertheless, oxidative modification by peroxyl radicals had no significant impact on the subunit structure of rice bran protein, but caused polypeptide chain aggregation of rice bran protein accompanied by partial rupture of the major polypeptide chain to form peptides. As AAPH concentration increased, rice bran protein solubility gradually decreased; however, water-holding capacity, oil-holding capacity, foaming activity, foaming stability, emulsifying activity,and emulsiong stability firstly increased and then decreased.
引文
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[19]ZHANG H J,ZHANG H,WANG L,et al.Preparation and functional properties of rice bran proteins from heat-stabilized defatted rice bran[J].Food Research International,2012,47(2):359-363.DOI:10.1016/j.foodres.2011.08.014.
[20]赵勇,万辉,郭善辉.对米糠油国标酸值等几个指标的探讨[J].粮食与食品工业,2 0 1 1,1 8(3):5 1-5 4.D O I:1 0.3 9 6 9/j.issn.1672-5026.2011.03.018.
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[22]陆玉芹,颜明月,陈德慰,等.鱼类加工制品蛋白质氧化程度分析[J].食品科学,2015,36(19):55-59.DOI:10.7506/spkx1002-6630-201519010.
[23]WASZCZAK C,AKTER S,JACQUES S,et al.Oxidative posttranslational modifications of cysteine residues in plant signal transduction[J].Journal of Experimental Botany,2015,66(10):2923-2934.DOI:10.1093/jxb/erv084.
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[26]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.
[27]JORGE C A,LUZ MARIA M,MARCELO V.Application of ATRFTIRspectroscopy to the study of thermally induced changes in secondary structure of protein molecules in solid state[J].Biopolymers,2015,103(10):574-584.DOI:10.1002/bip.22664.
[28]SUN W Z,ZHOU F B,SUN D W,et al.Effect of oxidation on the emulsifying properties of myofibrillar proteins[J].Food and Bioprocess Technology,2013,6(7):1703-1712.DOI:10.1007/s11947-012-0823-8.
[29]JACKSON M,MANTSCH H H.The use and misuse of FTIRspectroscopy in the determination of protein structure[J].Critical Reviews in Biochemistry and Molecular Biology,1995,30(2):95-120.DOI:10.3109/10409239509085140.
[30]BOCKER U,OFSTAD R,BERTRAM H C,et al.Salt-induced changes in pork myofibrillar tissue investigated by FT-IR microspectroscopy and light microscopy[J].Journal of Agricultural and Food Chemistry,2006,54(18):6733-6740.DOI:10.1021/jf060178q.
[31]ZHOU F B,ZHAO M M,CUI C,et al.Influence of linoleic acidinduced oxidative modifications on physicochemical changes and in vitro digestibility of porcine myofibrillar proteins[J].LWTFood Science and Technology,2015,61(2):414-421.DOI:10.1016/j.lwt.2014.12.037.
[32]张敏,周梅,王长远.米糠4种蛋白质的提取与功能性质[J].食品科学,2013,31(1):18-21.
[33]吴伟,吴晓娟,蔡勇建,等.米糠贮藏期间米糠球蛋白的结构变化[J].食品科学,2017,38(1):252-257.DOI:10.7506/spkx1002-6630-201701042.
[34]吴伟,吴晓娟,蔡勇建,等.米糠贮藏期间米糠谷蛋白结构变化的研究[J].中国粮油学报,2017,32(7):98-102;120.DOI:10.3969/j.issn.1003-0174.2017.07.016.
[35]吴伟.蛋白质氧化对大豆蛋白结构和凝胶性质的影响[D].无锡:江南大学,2009:29-30.
[2]SHARIF M K,BUTT M S,ANJUM F M,et al.Rice bran:a novel functional ingredient[J].Critical Reviews in Food Science and Nutrition,2014,54(6):807-816.DOI:10.1080/10408398.2011.608586.
[3]HAN S W,CHEE K M,CHO S J.Nutritional quality of rice bran protein in comparison to animal and vegetable protein[J].Food Chemistry,2015,172(3):766-769.DOI:10.1016/j.foodchem.2014.09.127.
[4]WANG C Y,XU F,LI D,et al.Physico-chemical and structural properties of four rice bran protein fractions based on the multiple solvent extraction method[J].Czech Journal of Food Sciences,2015,33(3):283-291.DOI:10.17221/462/2014-CJFS.
[5]杨祎晨,肖志刚,刘海飞,等.挤压法钝化米糠脂肪水解酶效果研究[J].中国食品学报,2014,14(9):90-99.
[6]KIM S M,CHUNG H J,LIM S T.Effect of various heat treatments on rancidity and some bioactive compounds of rice bran[J].Journal of Cereal Science,2014,60(1):243-248.DOI:10.1016/j.jcs.2014.04.001.
[7]SHACTER E.Quantification and significance of protein oxidation in biological samples[J].Drug Metabolism Reviews,2000,32:307-326.DOI:10.1081/DMR-100102336.
[8]吴伟,蔡勇建,吴晓娟,等.米糠贮藏时间对米糠蛋白结构的影响[J].现代食品科技,2017,33(1):173-178;205.DOI:10.13982/j.mfst.1673-9078.2017.1.027.
[9]蔡勇建,叶建芬,吴晓娟,等.米糠贮藏时间对米糠蛋白功能性质的影响[J].粮食与油脂,2015,28(6):31-34.DOI:10.3969/j.issn.1008-9578.2015.06.009.
[10]何莉媛,吴伟,吴晓娟,等.米糠贮藏时间对米糠蛋白体外胃蛋白酶消化性质及其消化产物抗氧化性的影响[J].中国油脂,2017,42(11):70-74.
[11]WERBER J,WANG Y J,MILLIGAN M,et al.Analysis of2,2’-azobis(2-amidinopropane)dihydrochloride degradation and hydrolysis in aqueous solutions[J].Journal of Pharmaceutical Sciences,2011,100(8):3307-3315.DOI:10.1002/jps.22578.
[12]WU W,ZHANG C M,KONG X Z,et al.Oxidative modification of soy protein by peroxyl radicals[J].Food Chemistry,2009,116(1):295-301.DOI:10.1016/j.foodchem.2009.02.049.
[13]YE L,LIAO Y,ZHAO M M,et al.Effect of protein oxidation on the conformational properties of peanut protein isolate[J].Journal of Chemistry,2013:1-6.DOI:10.1155/2013/423254.
[14]SUN W Z,CUI C,ZHAO M M,et al.Effects of composition and oxidation of proteins on their solubility,aggregation and proteolytic susceptibility during processing of cantonese sausage[J].Food Chemistry,2011,124(1):336-341.DOI:10.1016/j.foodchem.2010.06.042.
[15]CUI X H,XIONG Y L,KONG B H,et al.Hydroxyl radical-stressed whey protein isolate:chemical and structural properties[J].Food and Bioprocess Technology,2012,5(6):2454-2461.DOI:10.1007/s11947-011-0515-9.
[16]LIU Y L,LI X H,ZHOU X L,et al.Effects of glutaminase deamidation on the structure and solubility of rice glutelin[J].LWT-Food Science and Technology,2011,44(10):2205-2210.DOI:10.1016/j.lwt.2011.05.011.
[17]BENELHADI S,GHARSALLAOUI A,DEGRAEVE P,et al.Effect of pH on the functional properties of Arthrospira(Spirulina)platensis protein isolate[J].Food Chemistry,2016,194(1):1056-1063.DOI:10.1016/j.foodchem.2015.08.133.
[18]ZHU S M,LIN S L,RAMASWAMY H S,et al.Enhancement of functional properties of rice bran proteins by high pressure treatment and their correlation with surface hydrophobicity[J].Food and Bioprocess Technology,2017,10(2):317-327.DOI:10.1007/s11947-016-1818-7.
[19]ZHANG H J,ZHANG H,WANG L,et al.Preparation and functional properties of rice bran proteins from heat-stabilized defatted rice bran[J].Food Research International,2012,47(2):359-363.DOI:10.1016/j.foodres.2011.08.014.
[20]赵勇,万辉,郭善辉.对米糠油国标酸值等几个指标的探讨[J].粮食与食品工业,2 0 1 1,1 8(3):5 1-5 4.D O I:1 0.3 9 6 9/j.issn.1672-5026.2011.03.018.
[21]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.DOI:10.1016/j.meatsci.2013.12.001.
[22]陆玉芹,颜明月,陈德慰,等.鱼类加工制品蛋白质氧化程度分析[J].食品科学,2015,36(19):55-59.DOI:10.7506/spkx1002-6630-201519010.
[23]WASZCZAK C,AKTER S,JACQUES S,et al.Oxidative posttranslational modifications of cysteine residues in plant signal transduction[J].Journal of Experimental Botany,2015,66(10):2923-2934.DOI:10.1093/jxb/erv084.
[24]HEADLAM H A,DAVIES M J.Markers of protein oxidation:different oxidants give rise to variable yields of bound and released carbonyl products[J].Free Radical Biology and Medicine,2004,36(9):1175-1184.DOI:10.1016/j.freeradbiomed.2004.02.017.
[25]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.
[26]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.
[27]JORGE C A,LUZ MARIA M,MARCELO V.Application of ATRFTIRspectroscopy to the study of thermally induced changes in secondary structure of protein molecules in solid state[J].Biopolymers,2015,103(10):574-584.DOI:10.1002/bip.22664.
[28]SUN W Z,ZHOU F B,SUN D W,et al.Effect of oxidation on the emulsifying properties of myofibrillar proteins[J].Food and Bioprocess Technology,2013,6(7):1703-1712.DOI:10.1007/s11947-012-0823-8.
[29]JACKSON M,MANTSCH H H.The use and misuse of FTIRspectroscopy in the determination of protein structure[J].Critical Reviews in Biochemistry and Molecular Biology,1995,30(2):95-120.DOI:10.3109/10409239509085140.
[30]BOCKER U,OFSTAD R,BERTRAM H C,et al.Salt-induced changes in pork myofibrillar tissue investigated by FT-IR microspectroscopy and light microscopy[J].Journal of Agricultural and Food Chemistry,2006,54(18):6733-6740.DOI:10.1021/jf060178q.
[31]ZHOU F B,ZHAO M M,CUI C,et al.Influence of linoleic acidinduced oxidative modifications on physicochemical changes and in vitro digestibility of porcine myofibrillar proteins[J].LWTFood Science and Technology,2015,61(2):414-421.DOI:10.1016/j.lwt.2014.12.037.
[32]张敏,周梅,王长远.米糠4种蛋白质的提取与功能性质[J].食品科学,2013,31(1):18-21.
[33]吴伟,吴晓娟,蔡勇建,等.米糠贮藏期间米糠球蛋白的结构变化[J].食品科学,2017,38(1):252-257.DOI:10.7506/spkx1002-6630-201701042.
[34]吴伟,吴晓娟,蔡勇建,等.米糠贮藏期间米糠谷蛋白结构变化的研究[J].中国粮油学报,2017,32(7):98-102;120.DOI:10.3969/j.issn.1003-0174.2017.07.016.
[35]吴伟.蛋白质氧化对大豆蛋白结构和凝胶性质的影响[D].无锡:江南大学,2009:29-30.
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