自由基氧化对猪肌球蛋白-醛类化合物吸附特性的影响
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摘要
采用FeCl3/抗坏血酸/H2O2羟自由基氧化体系模拟猪肌球蛋白氧化,研究不同H2O2浓度对肌球蛋白巯基总量、活性巯基、二级结构和表面疏水性的影响及与4种典型醛类风味物质间的相互作用。结果表明:H2O2浓度对蛋白质构象有显著影响,随着H2O2浓度逐渐增加,活性巯基含量显著下降(P<0.05),表面疏水性显著增加(P<0.05)。当H2O2浓度在0～5 mmol/L之间时,巯基总量、α-螺旋相对含量显著下降(P<0.05),蛋白质吸附能力显著增强(P<0.05);当H2O2浓度在5～10 mmol/L之间时,α-螺旋相对含量显著上升(P<0.05),蛋白质吸附能力显著减弱(P<0.05);当H2O2浓度在10～20 mmol/L之间时,蛋白质吸附能力显著增强(P<0.05)。肌球蛋白与醛类化合物间的作用力主要为氢键和疏水相互作用,氢键和(或)疏水相互作用越强,蛋白质对醛类化合物吸附能力越强。
The effect of oxidation on protein conformation and binding capacity of porcine myosin were investigated in vitro using a FeCl3/Asc/H2 O2 hydroxyl radical oxidation system. The contents of total and reactive sulfhydryl groups, secondary structure and surface hydrophobicity were detected as a function of H2 O2 concentration, and the interactions between porcine myosin and four typical aldehyde compounds responsible for the flavor were investigated. It was demonstrated that the protein conformation was significantly affected by H2 O2 concentration. The content of reactive sulfhydryl groups was significantly decreased while surface hydrophobicity was significantly increased with the increase of H2 O2 concentration(P < 0.05). The contents of total sulfhydryl groups and α-helix were significantly decreased(P < 0.05) while the binding capacity between myosin and aldehyde compounds was significantly increased in the H2 O2 concentration range from 0 to5 mmol/L(P < 0.05). The α-helix content was significantly increased while the binding capacity of myosin was significantly decreased with increasing H2 O2 concentration to 10 mmol/L(P < 0.05). The binding capacity of myosin was significantly increased again with further increasing H2 O2 concentration to 20 mmol/L(P < 0.05). The main binding forces between aldehyde compounds and myosin were hydrogen bond and hydrophobic interaction. Stronger hydrogen bond and/or hydrophobic interaction led to a higher binding capacity of myosin to aldehyde compounds.
The effect of oxidation on protein conformation and binding capacity of porcine myosin were investigated in vitro using a FeCl3/Asc/H2 O2 hydroxyl radical oxidation system. The contents of total and reactive sulfhydryl groups, secondary structure and surface hydrophobicity were detected as a function of H2 O2 concentration, and the interactions between porcine myosin and four typical aldehyde compounds responsible for the flavor were investigated. It was demonstrated that the protein conformation was significantly affected by H2 O2 concentration. The content of reactive sulfhydryl groups was significantly decreased while surface hydrophobicity was significantly increased with the increase of H2 O2 concentration(P < 0.05). The contents of total sulfhydryl groups and α-helix were significantly decreased(P < 0.05) while the binding capacity between myosin and aldehyde compounds was significantly increased in the H2 O2 concentration range from 0 to5 mmol/L(P < 0.05). The α-helix content was significantly increased while the binding capacity of myosin was significantly decreased with increasing H2 O2 concentration to 10 mmol/L(P < 0.05). The binding capacity of myosin was significantly increased again with further increasing H2 O2 concentration to 20 mmol/L(P < 0.05). The main binding forces between aldehyde compounds and myosin were hydrogen bond and hydrophobic interaction. Stronger hydrogen bond and/or hydrophobic interaction led to a higher binding capacity of myosin to aldehyde compounds.
引文
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[9]SHAHIDI F.Flavor of meat,meat products and seafood[M].US:Springer,1998:1-3.
[10]SUPPAVORASATIT I.Deamidation of soy protein by proteinglutaminase:process evaluation and effect of deamidation on protein functional properties and flavor-protein interactions[J].Dissertations&Theses-Gradworks,2012,39(12):775-777.
[11]TOLDRáF,FLORES M.The role of muscle proteases and lipases in flavor development during the processing of dry-cured ham[J].Critical Reviews in Food Science&Nutrition,1998,38(4):331-352.DOI:10.1080/10408699891274237.
[12]吕彤,林俊杰,周昌瑜,等.热处理强度对猪肉肌球蛋白结构及风味成分吸附特性的影响[J].农业工程学报,2016,32(8):285-291.DOI:10.11975/j.issn.1002-6819.2016.08.040.
[13]周昌瑜,蒋娅婷,曹锦轩,等.热处理时间对鸭肉肌原纤维蛋白凝胶的风味吸附能力影响[J].现代食品科技,2016,32(3):218-224.
[14]YANG Q L,LOU X W,WANG Y,et al.Effect of pH on the interaction of volatile compounds with the myofibrillar proteins of duck meat[J].Poultry Science,2017,96(6):1963-1969.DOI:10.3382/ps/pew413.
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[17]HAMADA M,ISHIZAKI S,NAGAI T.Variation of SH content and kamaboko-gel forming ability of shark muscle protein by electrolysis[J].Journal of the Shimonoseki University of Fisheries,1994,42(42):131-135.
[18]ALIX A,PEDANOU G,BERJOT M.Fast determination of the quantitative secondary structure of proteins by using some parameters of the raman amide I band[J].Journal of Molecular Structure,1988,174(174):159-164.DOI:10.1016/0022-2860(88)80151-0.
[19]SUSI H,BYLER D M.Fourier deconvolution of the amide I Raman band of proteins as related to conformation[J].Applied Spectroscopy,1988,42(5):819-826.DOI:10.1366/0003702884428905.
[20]BOYER C,JOANDEL S,OUALI A,et al.Determination of surface hydrophobicity of fast and slow myosins from rabbit skeletal muscles:implication in heat-induced gelation[J].Journal of the Science of Food and Agriculture,1996,72(3):367-375.DOI:10.1002/(SICI)1097-0010(199611)72:3<367::AID-JSFA668>3.0.CO;2-D.
[21]李艳青,孔保华,杨赫鸿,等.自由基氧化引起鲤鱼肌原纤维蛋白结构的变化[J].食品科学,2012,33(13):70-74.
[22]SHARP J S,BECKER J M,HETTICH R L.Analysis of protein solvent accessible surfaces by photochemical oxidation and mass spectrometry[J].Analytical Chemistry,2004,76(3):672-683.DOI:10.1021/ac0302004.
[23]李银,孙红梅,张春晖,等.牛肉解冻过程中蛋白质氧化效应分析[J].中国农业科学,2013,46(7):1426-1433.DOI:10.3864/j.issn.0578-1752.2013.07.013.
[24]贾丹.青鱼肌肉蛋白质及其凝胶特性的研究[D].武汉:华中农业大学,2016:3-5.
[25]LI C,XIONG Y L.Disruption of secondary structure by oxidative stress alters the cross-linking pattern of myosin by microbial transglutaminase[J].Meat Science,2015,108(5):97-105.DOI:10.1016/j.meatsci.2015.05.024.
[26]OOIZUMI T,XIONG Y L.Identification of cross-linking site(s)of myosin heavy chains in oxidatively stressed chicken myofibrils[J].Journal of Food Science,2006,71(3):196-199.DOI:10.1111/j.1365-2621.2006.tb15617.x.
[27]LI C,XIONG Y L,CHEN J.Oxidation-induced unfolding facilitates myosin cross-linking in myofibrillar protein by microbial transglutaminase[J].Journal of Agricultural&Food Chemistry,2012,60(32):8020-8027.DOI:10.1021/jf302150h.
[28]TORNBERG E.Effects of heat on meat proteins-implications on structure and quality of meat products[J].Meat Science,2005,70(3):493-508.DOI:10.1016/j.meatsci.2004.11.021.
[29]BOURASSA P,KANAKIS C D,TARANTILIS P,et al.Resveratrol,genistein,and curcumin bind bovine serum albumin[J].Journal of Physical Chemistry B,2010,114(9):3348-3354.DOI:10.1021/jp9115996.
[30]AHMED-OUAMEUR A,DIAMANTOGLOU S,SEDAGHATHERATIM R,et al.The effects of drug complexation on the stability and conformation of human serum albumin[J].Cell Biochemistry&Biophysics,2006,45(2):203-213.
[31]NEAULT J F,NOVETTADELEN A,ARAKAWA H,et al.The effect of aspirin-HSA complexation on the protein secondary struct[J].Canadian Journal of Chemistry,2011,78(2):291-296.
[32]NEAULT J F,TAJMIR-RIAHI H A.Interaction of cisplatin with human serum albumin.Drug binding mode and protein secondary structure[J].Biochimica et Biophysica Acta,1998,1384(1):153-159.DOI:10.1016/S0167-4838(98)00011-9.
[33]CHANARAT S,BENJAKUL S,XIONG Y L.Physicochemical changes of myosin and gelling properties of washed tilapia mince as influenced by oxidative stress and microbial transglutaminase[J].Journal of Food Science&Technology,2015,52(6):3824-3836.DOI:10.1007/s13197-014-1448-x.
[34]WEEL K G C,BOELRIJK A E M,BURGER J J,et al.Effect of whey protein on the in vivo release of aldehydes[J].Journal of Agricultural&Food Chemistry,2003,51(16):4746-4752.
[35]TAN Y,SIEBERT K J.Modeling bovine serum albumin binding of flavor compounds(alcohols,aldehydes,esters,and ketones)as a function of molecular properties[J].Journal of Food Science,2008,73(1):56-63.DOI:10.1111/j.1750-3841.2007.00591.x.
[36]BREWER M S,VEGA J D.Detectable odor thresholds of selected lipid oxidation compounds in a meat model system[J].Journal of Food Science,2006,60(3):592-595.DOI:10.1111/j.1365-2621.1995.tb09834.x.
[2]陈洪生,孔保华,刁静静,等.氧化引起的肌肉蛋白质功能性变化及其控制技术的研究进展[J].食品科学,2015,36(11):215-220.DOI:10.7506/spkx1002-6630-201511041.
[3]STADTMAN E R.Oxidation of free amino acids and amino acid residues in proteins by radiolysis and by metal-catalyzed reactions[J].Annual Review of Biochemistry,1993,62(1):797-821.DOI:10.1146/annurev.bi.62.070193.004053.
[4]DAVIES M J.The oxidative environment and protein damage[J].Biochimica et Biophysica Acta,2005,1703(2):93-94.DOI:10.1016/j.bbapap.2004.08.007.
[5]STADTMAN E R.Protein oxidation in aging and age-related diseases[J].Annals of the New York Academy of Sciences,2001,928(1):22-38.DOI:10.1111/j.1749-6632.2001.tb05632.x.
[6]GIULIVI C,DAVIES K J.Dityrosine and tyrosine oxidation products are endogenous markers for the selective proteolysis of oxidatively modified red blood cell hemoglobin by(the 19S)proteasome[J].Journal of Biological Chemistry,1993,268(12):8752-8759.
[7]SHEN H R,SPIKES J D,KOPE菶KOV?P,et al.Photodynamic crosslinking of proteins.I.Model studies using histidine-and lysinecontaining N-(2-hydroxypropyl)methacrylamide copolymers[J].Journal of Photochemistry&Photobiology B Biology,1996,34(2/3):203-210.DOI:10.1016/1011-1344(96)07286-7.
[8]DAVIES K J A,DELSIGNORE M E.Protein damage and degradation by oxygen radicals.III.Modification of secondary and tertiary structure[J].Journal of Biological Chemistry,1987,262(20):9908-9913.
[9]SHAHIDI F.Flavor of meat,meat products and seafood[M].US:Springer,1998:1-3.
[10]SUPPAVORASATIT I.Deamidation of soy protein by proteinglutaminase:process evaluation and effect of deamidation on protein functional properties and flavor-protein interactions[J].Dissertations&Theses-Gradworks,2012,39(12):775-777.
[11]TOLDRáF,FLORES M.The role of muscle proteases and lipases in flavor development during the processing of dry-cured ham[J].Critical Reviews in Food Science&Nutrition,1998,38(4):331-352.DOI:10.1080/10408699891274237.
[12]吕彤,林俊杰,周昌瑜,等.热处理强度对猪肉肌球蛋白结构及风味成分吸附特性的影响[J].农业工程学报,2016,32(8):285-291.DOI:10.11975/j.issn.1002-6819.2016.08.040.
[13]周昌瑜,蒋娅婷,曹锦轩,等.热处理时间对鸭肉肌原纤维蛋白凝胶的风味吸附能力影响[J].现代食品科技,2016,32(3):218-224.
[14]YANG Q L,LOU X W,WANG Y,et al.Effect of pH on the interaction of volatile compounds with the myofibrillar proteins of duck meat[J].Poultry Science,2017,96(6):1963-1969.DOI:10.3382/ps/pew413.
[15]PéREZ-JUAN M,FLORES M,TOLDRáF.Effect of pork meat proteins on the binding of volatile compounds[J].Food Chemistry,2008,108(4):1226-1233.DOI:10.1016/j.foodchem.2007.04.037.
[16]GUICHARD E.Flavour retention and release from protein solutions[J].Biotechnology Advances,2006,24(2):226-229.DOI:10.1016/j.biotechadv.2005.11.003.
[17]HAMADA M,ISHIZAKI S,NAGAI T.Variation of SH content and kamaboko-gel forming ability of shark muscle protein by electrolysis[J].Journal of the Shimonoseki University of Fisheries,1994,42(42):131-135.
[18]ALIX A,PEDANOU G,BERJOT M.Fast determination of the quantitative secondary structure of proteins by using some parameters of the raman amide I band[J].Journal of Molecular Structure,1988,174(174):159-164.DOI:10.1016/0022-2860(88)80151-0.
[19]SUSI H,BYLER D M.Fourier deconvolution of the amide I Raman band of proteins as related to conformation[J].Applied Spectroscopy,1988,42(5):819-826.DOI:10.1366/0003702884428905.
[20]BOYER C,JOANDEL S,OUALI A,et al.Determination of surface hydrophobicity of fast and slow myosins from rabbit skeletal muscles:implication in heat-induced gelation[J].Journal of the Science of Food and Agriculture,1996,72(3):367-375.DOI:10.1002/(SICI)1097-0010(199611)72:3<367::AID-JSFA668>3.0.CO;2-D.
[21]李艳青,孔保华,杨赫鸿,等.自由基氧化引起鲤鱼肌原纤维蛋白结构的变化[J].食品科学,2012,33(13):70-74.
[22]SHARP J S,BECKER J M,HETTICH R L.Analysis of protein solvent accessible surfaces by photochemical oxidation and mass spectrometry[J].Analytical Chemistry,2004,76(3):672-683.DOI:10.1021/ac0302004.
[23]李银,孙红梅,张春晖,等.牛肉解冻过程中蛋白质氧化效应分析[J].中国农业科学,2013,46(7):1426-1433.DOI:10.3864/j.issn.0578-1752.2013.07.013.
[24]贾丹.青鱼肌肉蛋白质及其凝胶特性的研究[D].武汉:华中农业大学,2016:3-5.
[25]LI C,XIONG Y L.Disruption of secondary structure by oxidative stress alters the cross-linking pattern of myosin by microbial transglutaminase[J].Meat Science,2015,108(5):97-105.DOI:10.1016/j.meatsci.2015.05.024.
[26]OOIZUMI T,XIONG Y L.Identification of cross-linking site(s)of myosin heavy chains in oxidatively stressed chicken myofibrils[J].Journal of Food Science,2006,71(3):196-199.DOI:10.1111/j.1365-2621.2006.tb15617.x.
[27]LI C,XIONG Y L,CHEN J.Oxidation-induced unfolding facilitates myosin cross-linking in myofibrillar protein by microbial transglutaminase[J].Journal of Agricultural&Food Chemistry,2012,60(32):8020-8027.DOI:10.1021/jf302150h.
[28]TORNBERG E.Effects of heat on meat proteins-implications on structure and quality of meat products[J].Meat Science,2005,70(3):493-508.DOI:10.1016/j.meatsci.2004.11.021.
[29]BOURASSA P,KANAKIS C D,TARANTILIS P,et al.Resveratrol,genistein,and curcumin bind bovine serum albumin[J].Journal of Physical Chemistry B,2010,114(9):3348-3354.DOI:10.1021/jp9115996.
[30]AHMED-OUAMEUR A,DIAMANTOGLOU S,SEDAGHATHERATIM R,et al.The effects of drug complexation on the stability and conformation of human serum albumin[J].Cell Biochemistry&Biophysics,2006,45(2):203-213.
[31]NEAULT J F,NOVETTADELEN A,ARAKAWA H,et al.The effect of aspirin-HSA complexation on the protein secondary struct[J].Canadian Journal of Chemistry,2011,78(2):291-296.
[32]NEAULT J F,TAJMIR-RIAHI H A.Interaction of cisplatin with human serum albumin.Drug binding mode and protein secondary structure[J].Biochimica et Biophysica Acta,1998,1384(1):153-159.DOI:10.1016/S0167-4838(98)00011-9.
[33]CHANARAT S,BENJAKUL S,XIONG Y L.Physicochemical changes of myosin and gelling properties of washed tilapia mince as influenced by oxidative stress and microbial transglutaminase[J].Journal of Food Science&Technology,2015,52(6):3824-3836.DOI:10.1007/s13197-014-1448-x.
[34]WEEL K G C,BOELRIJK A E M,BURGER J J,et al.Effect of whey protein on the in vivo release of aldehydes[J].Journal of Agricultural&Food Chemistry,2003,51(16):4746-4752.
[35]TAN Y,SIEBERT K J.Modeling bovine serum albumin binding of flavor compounds(alcohols,aldehydes,esters,and ketones)as a function of molecular properties[J].Journal of Food Science,2008,73(1):56-63.DOI:10.1111/j.1750-3841.2007.00591.x.
[36]BREWER M S,VEGA J D.Detectable odor thresholds of selected lipid oxidation compounds in a meat model system[J].Journal of Food Science,2006,60(3):592-595.DOI:10.1111/j.1365-2621.1995.tb09834.x.
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