发酵对酸肉蛋白质结构的影响
|
摘要
以猪背最长肌为原料制备酸肉,研究蛋白质结构在发酵中的变化。结果表明,随发酵时间延长,蛋白质静电作用力逐渐减弱,疏水相互作用和二硫键作用显著增强后呈小幅度波动变化。蛋白紫外吸收结果显示,肌原纤维蛋白芳香族氨基酸残基偏向更加疏水的环境移动,肌浆蛋白酪氨酸、色氨酸残基微环境极性增加。内源荧光图谱显示,荧光强度总体呈下降趋势且相对发酵0 d时发生红移,表明色氨酸发生了氧化降解,其残基主要暴露到极性环境中。拉曼光谱分析表明,随发酵进行,蛋白质α-螺旋减少,β-折叠增多,发酵110 d酸肉蛋白二级结构可能发生了重排现象。结果表明长时间发酵可使酸肉蛋白质二级结构和三级结构发生变化,这些变化可从蛋白质结构水平上解释发酵对酸肉品质的影响。
Sour pork was prepared from pork longissimus dorsi muscle and the structural changes of proteins were studied during the fermentation process of sour pork. The results showed that the electrostatic force of proteins decreased with fermentation time, while the hydrophobic interaction and the disulfide bonding increased significantly followed by small fluctuations. The results of ultraviolet absorption spectroscopy showed that the aromatic amino acid residues of myofibrillar proteins were shifted to a more hydrophobic environment, and the polarity of the microenvironment around sarcoprotein tyrosine and tryptophan residues increased. The endogenous fluorescence spectra showed that the fluorescence intensity decreased generally, and red shift occurred when compared with before fermentation, which indicates that tryptophan underwent oxidative degradation, and its residues were mainly exposed to the polar environment. Raman analysis showed that the α-helical content decreased, while the β-sheet content increased gradually, and the secondary structure of proteins was reorganized after 110 days of fermentation. These results showed that long-time fermentation could change the secondary and tertiary structures of proteins in sour pork, which explains the effect of fermentation on sour pork quality at the level of protein structure.
Sour pork was prepared from pork longissimus dorsi muscle and the structural changes of proteins were studied during the fermentation process of sour pork. The results showed that the electrostatic force of proteins decreased with fermentation time, while the hydrophobic interaction and the disulfide bonding increased significantly followed by small fluctuations. The results of ultraviolet absorption spectroscopy showed that the aromatic amino acid residues of myofibrillar proteins were shifted to a more hydrophobic environment, and the polarity of the microenvironment around sarcoprotein tyrosine and tryptophan residues increased. The endogenous fluorescence spectra showed that the fluorescence intensity decreased generally, and red shift occurred when compared with before fermentation, which indicates that tryptophan underwent oxidative degradation, and its residues were mainly exposed to the polar environment. Raman analysis showed that the α-helical content decreased, while the β-sheet content increased gradually, and the secondary structure of proteins was reorganized after 110 days of fermentation. These results showed that long-time fermentation could change the secondary and tertiary structures of proteins in sour pork, which explains the effect of fermentation on sour pork quality at the level of protein structure.
引文
[1]顾书媛,余静,莫玲,等.蛋白质氧化及其对机体氧化还原状态的影响[J].食品工业科技,2012,33(17):382-387.DOI:10.13386/j.issn1002-0306.2012.17.009.
[2]ESTéVEZ M.Protein carbonyls in meat systems:a review[J].Meat Science,2011,89:259-279.DOI:10.1016/j.meatsci.2011.04.025.
[3]李华丽.酸肉生产主发酵期发酵条件的确定[J].中国食物与营养,2005(4):41-43.DOI:10.3969/j.issn.1006-9577.2005.04.013.
[4]李宗军,江汉湖.响应曲面法评价环境因子对肉品发酵过程中微生物的影响[J].食品科学,2004,25(1):39-44.DOI:10.3321/j.issn:1002-6630.2004.01.004.
[5]余冰,周红丽,李宗军.固相微萃取分析发酵肉制品中的挥发性风味组分[J].湖南农业大学学报(自然科学版),2007,33(2):232-234.DOI:10.3321/j.issn:1007-1032.2007.02.026.
[6]周才琼,李艳芳,杜木英.渝黔地区传统酸肉发酵过程中微生物区系研究[J].食品工业科技,2010,31(4):171-175.DOI:10.13386/j.issn1002-0306.2010.04.035.
[7]周才琼,陈东华,杜木英.酸肉发酵中蛋白质降解及影响因素的研究[J].食品科学,2009,30(7):127-130.DOI:10.3321/j.issn:1002-6630.2009.07.029.
[8]李成龙,袁军,刘淑贞,等.米粉及盐添加量对发酵酸肉蛋白质降解及GABA形成的影响[J].食品科学,2015,36(13):202-206.DOI:10.7506/spkx1002-6630-201513037.
[9]CLAUSEN I,JAKOBSEN M,ERTBJERG P,et al.Modified atmosphere packaging affects lipid oxidation,myofibrillar fragmentation index and eating quality of beef[J].Packaging Technology&Science,2009,22(2):85-96.DOI:10.1002/pts.828.
[10]SUN W,CUI C,ZHAO 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.
[11]CARDAMONE M,PURI N K.Spectrofluorimetric assessment of the surface hydrophobicity of proteins[J].Biochemical Journal,1992,282(2):589.DOI:10.1042/bj2820589.
[12]TRAORE S,AUBRY L,GATELLIER P,et al.Higher drip loss is associated with protein oxidation.[J].Meat Science,2012,90(4):917.DOI:10.1016/j.meatsci.2011.11.033.
[13]VISESSANGUAN W,BENJAKUL S,RIEBROY S,et al.Changes in composition and functional properties of proteins and their contributions to Nham characteristics[J].Meat Science,2004,66(3):579-588.DOI:10.1016/S0309-1740(03)00172-4.
[14]LOBO F,VENTANAS S,MORCUENDE D,et al.Underlying chemical mechanisms of the contradictory effects of NaCl reduction on the redox-state of meat proteins in fermented sausages[J].LWT-Food Science and Technology,2016,69:110-116.DOI:10.1016/j.lwt.2016.01.047.
[15]任丽娜.白鲢鱼肉肌原纤维蛋白冷冻变性的研究[D].无锡:江南大学,2014.
[16]ZAJ?C A,HANUZA J,DYMI?SKA L.Raman spectroscopy in determination of horse meat content in the mixture with other meats[J].Food Chemistry,2014,156(3):333.DOI:10.1016/j.foodchem.2014.02.002.
[17]XU Y,XIA W,YANG F,et al.Protein molecular interactions involved in the gel network formation of fermented silver carp mince inoculated with Pediococcus pentosaceus[J].Food Chemistry,2010,120(3):717-723.DOI:10.1016/j.foodchem.2009.10.068.
[18]VISSCHERS R W,DE JONGH H H.Disulphide bond formation in food protein aggregation and gelation[J].Biotechnology Advances,2005,23(1):75-80.DOI:10.1016/j.biotechadv.2004.09.005.
[19]SOLADOYE O P,JUáREZ M L,AALHUS J L,et al.Protein oxidation in processed meat:mechanisms and potential implications on human health[J].Comprehensive Reviews in Food Science&Food Safety,2015,14(2):106-122.DOI:10.1111/1541-4337.12127.
[20]SKJERVOLD P O,TAYLOR R G,WOLD J P,et al.Development of intrinsic fluorescent multispectral imagery specific for fat,connective tissue,and myofibers in meat[J].Journal of Food Science,2010,68(4):1161-1168.DOI:10.1111/j.1365-2621.2003.tb09618.x.
[21]LIU R,ZHAO S M,XIE B J,et al.Contribution of protein conformation and intermolecular bonds to fish and pork gelation properties[J].Food Hydrocolloids,2011,25(5):898-906.DOI:10.1016/j.foodhyd.2010.08.016.
[22]HOWELL N,LICHAN E.Elucidation of interactions of lysozyme with whey proteins by Raman spectroscopy[J].International Journal of Food Science&Technology,1996,31(5):439-451.DOI:10.1046/j.1365-2621.1996.00364.x.
[23]BADII F,HOWELL N K.Fish gelatin:structure,gelling properties and interaction with egg albumen proteins[J].Food Hydrocolloids,2006,20(5):630-640.DOI:10.1016/j.foodhyd.2005.06.006.
[24]杨方.鱼肉内源酶对发酵鱼糜凝胶和抗氧化特性影响的研究[D].无锡:江南大学,2016.
[25]谢媚,曹锦轩,潘道东,等.滚揉对成熟过程中鹅肉品质及其蛋白质结构的影响[J].现代食品科技,2014(10):205-211.DOI:10.13982/j.mfst.1673-9078.2014.10.035.
[26]曹锦轩,张玉林,韩敏义,等.腊肉加工过程中肌原纤维蛋白结构的变化[J].中国农业科学,2013,46(18):3871-3877.DOI:10.3864/j.issn.0578-1752.2013.18.017.
[27]CARECHE M,LI-CHAN E C Y.Structural changes in cod myosin after modification with formaldehyde or frozen storage[J].Journal of Food Science,2006,62(4):717-723.DOI:10.1111/j.1365-2621.1997.tb15443.x.
[28]周绪霞,姜珊,顾赛麒,等.油茶籽油对鱼糜凝胶特性及凝胶结构的影响[J].食品科学,2017,38(9):27-33.DOI:10.7506/spkx1002-6630-201709005.
[29]秦影,汤海青,欧昌荣,等.超高压处理对大黄鱼鱼糜水分状态和蛋白质结构的影响[J].农业工程学报,2015,31(23):246-252.DOI:10.11975/j.issn.1002-6819.2015.23.033.
[30]HERRERO A M.Raman spectroscopy a promising technique for quality assessment of meat and fish:a review[J].Food Chemistry,2008,107(4):1642-1651.DOI:10.1016/j.foodchem.2007.10.014.
[31]张兴,杨玉玲,马云,等.pH对肌原纤维蛋白及其热诱导凝胶非共价键作用力与结构的影响[J].中国农业科学,2017,50(3):564-573.DOI:10.3864/j.issn.0578-1752.2017.03.015.
[32]李可,赵颖颖,康壮丽,等.NaCl对猪肉糜加工特性和蛋白质二级结构的影响[J].食品科学,2017,38(15):77-81.DOI:10.7506/spkx1002-6630-201715013.
[33]孙为正.广式腊肠加工过程中脂质水解、蛋白质降解及风味成分变化研究[D].广州:华南理工大学,2011.
[34]陈超,高学军,刘营,等.产蛋白酶乳酸茵的筛选及蛋白酶的纯化[J].乳业科学与技术,2008,31(4):160-162.DOI:10.15922/j.cnki.jdst.2008.04.003.
[2]ESTéVEZ M.Protein carbonyls in meat systems:a review[J].Meat Science,2011,89:259-279.DOI:10.1016/j.meatsci.2011.04.025.
[3]李华丽.酸肉生产主发酵期发酵条件的确定[J].中国食物与营养,2005(4):41-43.DOI:10.3969/j.issn.1006-9577.2005.04.013.
[4]李宗军,江汉湖.响应曲面法评价环境因子对肉品发酵过程中微生物的影响[J].食品科学,2004,25(1):39-44.DOI:10.3321/j.issn:1002-6630.2004.01.004.
[5]余冰,周红丽,李宗军.固相微萃取分析发酵肉制品中的挥发性风味组分[J].湖南农业大学学报(自然科学版),2007,33(2):232-234.DOI:10.3321/j.issn:1007-1032.2007.02.026.
[6]周才琼,李艳芳,杜木英.渝黔地区传统酸肉发酵过程中微生物区系研究[J].食品工业科技,2010,31(4):171-175.DOI:10.13386/j.issn1002-0306.2010.04.035.
[7]周才琼,陈东华,杜木英.酸肉发酵中蛋白质降解及影响因素的研究[J].食品科学,2009,30(7):127-130.DOI:10.3321/j.issn:1002-6630.2009.07.029.
[8]李成龙,袁军,刘淑贞,等.米粉及盐添加量对发酵酸肉蛋白质降解及GABA形成的影响[J].食品科学,2015,36(13):202-206.DOI:10.7506/spkx1002-6630-201513037.
[9]CLAUSEN I,JAKOBSEN M,ERTBJERG P,et al.Modified atmosphere packaging affects lipid oxidation,myofibrillar fragmentation index and eating quality of beef[J].Packaging Technology&Science,2009,22(2):85-96.DOI:10.1002/pts.828.
[10]SUN W,CUI C,ZHAO 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.
[11]CARDAMONE M,PURI N K.Spectrofluorimetric assessment of the surface hydrophobicity of proteins[J].Biochemical Journal,1992,282(2):589.DOI:10.1042/bj2820589.
[12]TRAORE S,AUBRY L,GATELLIER P,et al.Higher drip loss is associated with protein oxidation.[J].Meat Science,2012,90(4):917.DOI:10.1016/j.meatsci.2011.11.033.
[13]VISESSANGUAN W,BENJAKUL S,RIEBROY S,et al.Changes in composition and functional properties of proteins and their contributions to Nham characteristics[J].Meat Science,2004,66(3):579-588.DOI:10.1016/S0309-1740(03)00172-4.
[14]LOBO F,VENTANAS S,MORCUENDE D,et al.Underlying chemical mechanisms of the contradictory effects of NaCl reduction on the redox-state of meat proteins in fermented sausages[J].LWT-Food Science and Technology,2016,69:110-116.DOI:10.1016/j.lwt.2016.01.047.
[15]任丽娜.白鲢鱼肉肌原纤维蛋白冷冻变性的研究[D].无锡:江南大学,2014.
[16]ZAJ?C A,HANUZA J,DYMI?SKA L.Raman spectroscopy in determination of horse meat content in the mixture with other meats[J].Food Chemistry,2014,156(3):333.DOI:10.1016/j.foodchem.2014.02.002.
[17]XU Y,XIA W,YANG F,et al.Protein molecular interactions involved in the gel network formation of fermented silver carp mince inoculated with Pediococcus pentosaceus[J].Food Chemistry,2010,120(3):717-723.DOI:10.1016/j.foodchem.2009.10.068.
[18]VISSCHERS R W,DE JONGH H H.Disulphide bond formation in food protein aggregation and gelation[J].Biotechnology Advances,2005,23(1):75-80.DOI:10.1016/j.biotechadv.2004.09.005.
[19]SOLADOYE O P,JUáREZ M L,AALHUS J L,et al.Protein oxidation in processed meat:mechanisms and potential implications on human health[J].Comprehensive Reviews in Food Science&Food Safety,2015,14(2):106-122.DOI:10.1111/1541-4337.12127.
[20]SKJERVOLD P O,TAYLOR R G,WOLD J P,et al.Development of intrinsic fluorescent multispectral imagery specific for fat,connective tissue,and myofibers in meat[J].Journal of Food Science,2010,68(4):1161-1168.DOI:10.1111/j.1365-2621.2003.tb09618.x.
[21]LIU R,ZHAO S M,XIE B J,et al.Contribution of protein conformation and intermolecular bonds to fish and pork gelation properties[J].Food Hydrocolloids,2011,25(5):898-906.DOI:10.1016/j.foodhyd.2010.08.016.
[22]HOWELL N,LICHAN E.Elucidation of interactions of lysozyme with whey proteins by Raman spectroscopy[J].International Journal of Food Science&Technology,1996,31(5):439-451.DOI:10.1046/j.1365-2621.1996.00364.x.
[23]BADII F,HOWELL N K.Fish gelatin:structure,gelling properties and interaction with egg albumen proteins[J].Food Hydrocolloids,2006,20(5):630-640.DOI:10.1016/j.foodhyd.2005.06.006.
[24]杨方.鱼肉内源酶对发酵鱼糜凝胶和抗氧化特性影响的研究[D].无锡:江南大学,2016.
[25]谢媚,曹锦轩,潘道东,等.滚揉对成熟过程中鹅肉品质及其蛋白质结构的影响[J].现代食品科技,2014(10):205-211.DOI:10.13982/j.mfst.1673-9078.2014.10.035.
[26]曹锦轩,张玉林,韩敏义,等.腊肉加工过程中肌原纤维蛋白结构的变化[J].中国农业科学,2013,46(18):3871-3877.DOI:10.3864/j.issn.0578-1752.2013.18.017.
[27]CARECHE M,LI-CHAN E C Y.Structural changes in cod myosin after modification with formaldehyde or frozen storage[J].Journal of Food Science,2006,62(4):717-723.DOI:10.1111/j.1365-2621.1997.tb15443.x.
[28]周绪霞,姜珊,顾赛麒,等.油茶籽油对鱼糜凝胶特性及凝胶结构的影响[J].食品科学,2017,38(9):27-33.DOI:10.7506/spkx1002-6630-201709005.
[29]秦影,汤海青,欧昌荣,等.超高压处理对大黄鱼鱼糜水分状态和蛋白质结构的影响[J].农业工程学报,2015,31(23):246-252.DOI:10.11975/j.issn.1002-6819.2015.23.033.
[30]HERRERO A M.Raman spectroscopy a promising technique for quality assessment of meat and fish:a review[J].Food Chemistry,2008,107(4):1642-1651.DOI:10.1016/j.foodchem.2007.10.014.
[31]张兴,杨玉玲,马云,等.pH对肌原纤维蛋白及其热诱导凝胶非共价键作用力与结构的影响[J].中国农业科学,2017,50(3):564-573.DOI:10.3864/j.issn.0578-1752.2017.03.015.
[32]李可,赵颖颖,康壮丽,等.NaCl对猪肉糜加工特性和蛋白质二级结构的影响[J].食品科学,2017,38(15):77-81.DOI:10.7506/spkx1002-6630-201715013.
[33]孙为正.广式腊肠加工过程中脂质水解、蛋白质降解及风味成分变化研究[D].广州:华南理工大学,2011.
[34]陈超,高学军,刘营,等.产蛋白酶乳酸茵的筛选及蛋白酶的纯化[J].乳业科学与技术,2008,31(4):160-162.DOI:10.15922/j.cnki.jdst.2008.04.003.