超高压处理对养殖大黄鱼肌原纤维蛋白结构的影响
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摘要
本研究以养殖大黄鱼鱼肉为原料,经过超高压处理(压力:150、200、250、300、350、400、450、500 MPa;保压时间:5、10、15、20、25、30 min)后,通过荧光分光光度法测定肌原纤维蛋白的内源荧光强度,采用傅里叶变换红外光谱法测定肌原纤维蛋白官能团结构,采用圆二色光谱法测定蛋白二级结构变化,采用激光拉曼光谱法分析蛋白质的化学基团信息,采用扫描电子显微镜法分析蛋白微观结构变化。结果表明:养殖大黄鱼鱼肉肌原纤维蛋白的荧光及最大发射波长受处理压力和保压时间的影响而变化,随着压力的增大和保压时间的延长,最大发射波长峰值变大;超高压处理影响了肌原纤维蛋白的三级结构,其氨基酸残基所处化学环境极性增强;超高压处理影响了肌原纤维蛋白的二级结构,其α-螺旋含量随压力的升高和保压时间的延长而逐渐降低;超高压处理影响蛋白构象变化并引起蛋白变性,压力越大、保压时间越长,其变性程度越大,变性程度直接影响着肌原纤维蛋白的微观结构。养殖大黄鱼肌原纤维蛋白在超高压处理后结构发生了变化,其二级、三级结构均有改变并引起蛋白变性,鱼肉品质随之发生变化。
In this paper, large yellow croaker was subjected to ultra-high pressure treatment at pressures of 150, 200, 250,300, 350, 400, 450 and 500 MPa for different durations of 5, 10, 15, 20, 25 and 30 min for modification of myofibrillar proteins. The endogenous fluorescence intensity of myofibrillar proteins was determined by fluorescence spectrophotometry,the myofibrillar protein functional structure was determined by Fourier transform infrared spectroscopy, and the secondary structure was determined by circular dichroism spectroscopy. Raman spectroscopy was employed to determine the information about chemical groups in the proteins, and the microstructure changes were observed by scanning electron microscopy. The results showed that the fluorescence intensity and maximum emission wavelength of myofibrillar proteins were influenced by pressure and holding time, and the maximum fluorescence emission intensity increased with the increase in the variables. Ultra-high pressure treatment affected the tertiary structure of myofibrillar proteins and enhanced the polarity of the surrounding environment of amino acid residues. Ultra-high pressure also affected the secondary structure;the content of α-helix structure gradually decreased with the increase in pressure and holding time. Moreover, ultra-high pressure affected protein conformational changes and caused protein denaturation; the degree of denaturation, directly affecting the microstructure of myofibrillar proteins, was positively correlated with pressure and holding time. Conclusively,the secondary and tertiary structure of myofibrillar proteins from cultured large yellow croaker was changed after ultra-high pressure treatment, causing protein denaturation and accordingly quality changes of fish meat.
In this paper, large yellow croaker was subjected to ultra-high pressure treatment at pressures of 150, 200, 250,300, 350, 400, 450 and 500 MPa for different durations of 5, 10, 15, 20, 25 and 30 min for modification of myofibrillar proteins. The endogenous fluorescence intensity of myofibrillar proteins was determined by fluorescence spectrophotometry,the myofibrillar protein functional structure was determined by Fourier transform infrared spectroscopy, and the secondary structure was determined by circular dichroism spectroscopy. Raman spectroscopy was employed to determine the information about chemical groups in the proteins, and the microstructure changes were observed by scanning electron microscopy. The results showed that the fluorescence intensity and maximum emission wavelength of myofibrillar proteins were influenced by pressure and holding time, and the maximum fluorescence emission intensity increased with the increase in the variables. Ultra-high pressure treatment affected the tertiary structure of myofibrillar proteins and enhanced the polarity of the surrounding environment of amino acid residues. Ultra-high pressure also affected the secondary structure;the content of α-helix structure gradually decreased with the increase in pressure and holding time. Moreover, ultra-high pressure affected protein conformational changes and caused protein denaturation; the degree of denaturation, directly affecting the microstructure of myofibrillar proteins, was positively correlated with pressure and holding time. Conclusively,the secondary and tertiary structure of myofibrillar proteins from cultured large yellow croaker was changed after ultra-high pressure treatment, causing protein denaturation and accordingly quality changes of fish meat.
引文
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[12]CHIN K B,GO M Y,XIONG Youling L..Konjac flour improved textural and water retention properties of transglutaminase-mediated,heat-induced porcine myofibrillar protein gel:effect of salt level and transglutaminase incubation[J].Meat Science,2009,81(3):565-572.DOI:10.1016/j.meatsci.2008.10.012.
[13]XU Yangshun,XIA Wenshui,JIANG Qixing.Aggregation and structural changes of silver carp actomyosin as affected by mild acidification with D-gluconic acidδ-lactone[J].Food Chemistry,2012,134(2):1005-1010.DOI:10.1016/j.foodchem.2012.02.216.
[14]WELLS J A,WERBER M M,LEGG J I,et al.Inactivation of myosin subfragment one by cobalt(II)/cobalt(III)phenanthroline complexes.I.incorporation of Co(III)by in situ oxidation of Co(II)[J].Biochemistry,1979,18(22):4793-4799.DOI:10.1021/bi00589a005.
[15]MASAHIRO O,SOICHIRO N,YASUMI H,et al.Raman spectroscopic study of changes in fish Actomyosin during setting[J].Journal of Agricultural and Food Chemistry,1999,47(8):3309-3318.DOI:10.1021/jf9813079.
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[17]LEFEVRE F,FAUCONNEAU B,THOMPSON J W,et al.Thermal denaturation and aggregation properties of atlantic salmon myofibrils and myosin from white and red muscles[J].Journal of Agricultural and Food Chemistry,2007,55(12):4761-4770.DOI:10.1021/jf063045d.
[18]孔继烈,吴性良.分析化学原理[M].北京:化学工业出版社,2010:10-80.
[19]GREENFIELD N J.Using circular dichroism spectra to estimate protein secondary structure[J].Nature Protocols,2005,1(6):2876-2890.DOI:10.1038/nprot.2006.202.
[20]SREERAMA N,VENYAMINOV S Y U,WOODY R W.Estimation of the number of alpha-helical and beta-strand segments in proteins using circular dichroism spectroscopy[J].Protein Science,1999,8(2):370-380.DOI:10.1110/ps.8.2.370.
[21]ROBERT T,KUMOSINSKI T F,SERGE N T,et al.The circular dichroism of theβstructure of poly-L-lysine[J].Biochemical and Biophysical Research Communications,1966,23(2):163-169.DOI:10.1016/0006-291x(66)90522-5.
[22]MOZHAEV V V,LANGE R,KUDRYASHOVA E V,et al.Application of high hydrostatic pressure for increasing activity and stability of enzymes[J].Biotechnology and Bioengineering,1996,52(2):320-331.DOI:10.1002/(sici)1097-0290(19961020)52:2<320::aid-bit12>3.0.co;2-n.
[23]TAKESHI S,OHNO T,OTSUKA-FUCHINO H,et al.Carp natural actomyosin:thermal denaturation mechanism[J].Journal of Food Science,1994,59(5):1002-1008.DOI:10.1111/j.1365-2621.1994.tb08177.x.
[24]LI L K,SPECTOR A.Circular dichroism of beta-poly-L-lysine[J].Journal of the American Chemical Society,1969,91(1):220-222.DOI:10.1021/ja01029a058.
[25]KATO A,TSUTSUI N,MATSUDOMI N,et al.Effects of partial denaturation on surface properties of ovalbumin and lysozyme[J].Journal of the Agricultural and Biological Chemistry,1981,45(12):2755-2760.DOI:10.1080/00021369.1981.10864964.
[26]伍林,欧阳兆辉,曹淑超,等.拉曼光谱技术的应用及研究进展[J].光散射学报,2005,17(2):180-186.DOI:10.3969/j.issn.1004-5929.2005.02.013.
[27]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.
[28]CARECHE M,GARCIA M L,HERRERO A,et al.Structural properties of aggregates from frozen stored hake muscle proteins[J].Journal of Food Science,2002,67(8):2827-2832.DOI:10.1111/j.1365-2621.2002.tb08823.x.
[29]HERRERO A M.Raman spectroscopy for monitoring protein structure in muscle food systems[J].Critical Reviews in Food Science and Nutrition,2008,48(6):512-523.DOI:10.1080/10408390701537385.
[30]CHOI S M,MA C Y.Structural characterization of globulin from common buckwheat(Fagopyrum esculentum Moench)using circular dichroism and Raman spectroscopy[J].Food Chemistry,2007,102(1):150-160.DOI:10.1016/j.foodchem.2006.05.011.
[31]胡飞华.梅鱼鱼糜超高压凝胶化工艺及凝胶机理的研究[D].杭州:浙江工商大学,2010:22-40.
[32]ONWULATA C I,ISOBE S,TOMASULA P M,et al.Properties of whey protein isolates extruded under acidic and alkaline conditions[J].Journal of Dairy Science,2006,89(1):71-81.DOI:10.3168/jds.s0022-0302(06)72070-7.
[33]杨华,陆森超,张慧恩,等.超高压处理对养殖大黄鱼风味及品质的影响[J].食品科学,2014,35(16):244-249.DOI:10.7506/spkx1002-6630-201416047.
[34]付强.超高压处理对鲢鱼鱼糜品质特性的影响[D].上海:上海海洋大学,2015:21-80.
[35]仪淑敏,马兴胜,励建荣,等.超高压诱导鱼糜凝胶形成中水分特性及凝胶强度的相关性研究[J].中国食品学报,2015,15(7):26-31.DOI:10.16429/j.1009-7848.2015.07.004.
[36]刘坚,李艳红,缪铭,等.超高压对不同缓冲体系中鹰嘴豆分离蛋白溶解性的影响[J].食品工业科技,2007(11):90-92.DOI:10.13386/j.issn1002-0306.2007.11.017.
[37]王喜波,王健,张泽宇,等.物理改性对大豆蛋白柔性与乳化性的影响及其相关性分析[J].农业机械学报,2017,48(7):339-344.DOI:10.6041/j.issn.1000-1298.2017.07.043.
[38]郑捷,尚校兰,刘安军.超高压处理对海鲈鱼鱼肉凝胶形成作用[J].食品科学,2013,34(19):88-92.DOI:10.7506/spkx1002-6630-201319020.
[39]刘书成,邓倩琳,黄万有,等.超高压处理对凡纳滨对虾虾仁蛋白质和微观结构的影响[J].水产学报,2017,41(6):877-887.DOI:10.11964/jfc.20170110674.
[40]周敏,陆维克,陆金金,等.超高压处理对副溶血性弧菌胞内蛋白质的影响[J].食品科学,2017,38(1):99-104.DOI:10.7506/spkx1002-6630-201701016.
[2]肖雄,林淑琴,吴雄飞,等.三种不同养殖模式下大黄鱼鱼皮、鱼鳞挥发性风味成分分析[J].中国水产科学,2017,24(2):341-354.DOI:10.3724/SP.J.1118.2017.16140.
[3]陈博欧,杨正勇.中国养殖大黄鱼国际竞争力分析[J].中国渔业经济,2017,35(3):53-59.DOI:10.3969/j.issn.1009-590X.2017.03.008.
[4]陈世达,朱艳杰,张登科,等.超高压处理对养殖大黄鱼肉蛋白质特性的影响[J].食品安全质量检测学报,2017,8(2):579-586.DOI:10.3969/j.issn.2095-0381.2017.02.036.
[5]李婷婷.大黄鱼生物保鲜技术及新鲜度指示蛋白研究[D].杭州:浙江工商大学,2013:21-44.
[6]杨华,刘斌斌,娄永江.养殖大黄鱼脱脂脱腥技术的研究[J].食品科技,2012,37(6):154-157.DOI:10.7666/d.d013990.
[7]周果,杨文鸽,崔燕,等.超高压处理对三疣梭子蟹感官及其肌原纤维蛋白生化特性的影响[J].食品科学,2017,38(23):269-274.DOI:10.7506/spkx1002-6630-201723043.
[8]TRUONG B Q,BUCKOW R,STATHOPOULOS C E,et al.Advances in high-pressure processing of fish muscles[J].Food Engineering Reviews,2015,7(2):109-129.DOI:10.1007/s12393-014-9084-9.
[9]刘延奇,吴史博.超高压对食品品质的影响[J].食品研究与开发,2008(3):137-141.DOI:10.3969/j.issn.1005-6521.2008.03.045.
[10]ZHOU H,LIU B,YUE W L,et al.Research progress on application of ultra-high pressure technology in the production of pre prepared food[J].Packaging & Food Machinery,2017,2:47-51.DOI:10.1002/9781119962045.ch12.
[11]JIANG Xinjing,ZHANG Zhijun,CAI Huinong,et al.The effect of soybean trypsin inhibitor on the degradation of myofibrillar proteins by an endogenous serine proteinase of crucian carp[J].Food Chemistry,2006,94(4):498-503.DOI:10.1016/j.foodchem.2004.11.046.
[12]CHIN K B,GO M Y,XIONG Youling L..Konjac flour improved textural and water retention properties of transglutaminase-mediated,heat-induced porcine myofibrillar protein gel:effect of salt level and transglutaminase incubation[J].Meat Science,2009,81(3):565-572.DOI:10.1016/j.meatsci.2008.10.012.
[13]XU Yangshun,XIA Wenshui,JIANG Qixing.Aggregation and structural changes of silver carp actomyosin as affected by mild acidification with D-gluconic acidδ-lactone[J].Food Chemistry,2012,134(2):1005-1010.DOI:10.1016/j.foodchem.2012.02.216.
[14]WELLS J A,WERBER M M,LEGG J I,et al.Inactivation of myosin subfragment one by cobalt(II)/cobalt(III)phenanthroline complexes.I.incorporation of Co(III)by in situ oxidation of Co(II)[J].Biochemistry,1979,18(22):4793-4799.DOI:10.1021/bi00589a005.
[15]MASAHIRO O,SOICHIRO N,YASUMI H,et al.Raman spectroscopic study of changes in fish Actomyosin during setting[J].Journal of Agricultural and Food Chemistry,1999,47(8):3309-3318.DOI:10.1021/jf9813079.
[16]张延会,吴良平,孙真荣.拉曼光谱技术应用进展[J].化学教学,2006,4:32-35.DOI:10.3969/j.issn.1005-6629.2006.04.014.
[17]LEFEVRE F,FAUCONNEAU B,THOMPSON J W,et al.Thermal denaturation and aggregation properties of atlantic salmon myofibrils and myosin from white and red muscles[J].Journal of Agricultural and Food Chemistry,2007,55(12):4761-4770.DOI:10.1021/jf063045d.
[18]孔继烈,吴性良.分析化学原理[M].北京:化学工业出版社,2010:10-80.
[19]GREENFIELD N J.Using circular dichroism spectra to estimate protein secondary structure[J].Nature Protocols,2005,1(6):2876-2890.DOI:10.1038/nprot.2006.202.
[20]SREERAMA N,VENYAMINOV S Y U,WOODY R W.Estimation of the number of alpha-helical and beta-strand segments in proteins using circular dichroism spectroscopy[J].Protein Science,1999,8(2):370-380.DOI:10.1110/ps.8.2.370.
[21]ROBERT T,KUMOSINSKI T F,SERGE N T,et al.The circular dichroism of theβstructure of poly-L-lysine[J].Biochemical and Biophysical Research Communications,1966,23(2):163-169.DOI:10.1016/0006-291x(66)90522-5.
[22]MOZHAEV V V,LANGE R,KUDRYASHOVA E V,et al.Application of high hydrostatic pressure for increasing activity and stability of enzymes[J].Biotechnology and Bioengineering,1996,52(2):320-331.DOI:10.1002/(sici)1097-0290(19961020)52:2<320::aid-bit12>3.0.co;2-n.
[23]TAKESHI S,OHNO T,OTSUKA-FUCHINO H,et al.Carp natural actomyosin:thermal denaturation mechanism[J].Journal of Food Science,1994,59(5):1002-1008.DOI:10.1111/j.1365-2621.1994.tb08177.x.
[24]LI L K,SPECTOR A.Circular dichroism of beta-poly-L-lysine[J].Journal of the American Chemical Society,1969,91(1):220-222.DOI:10.1021/ja01029a058.
[25]KATO A,TSUTSUI N,MATSUDOMI N,et al.Effects of partial denaturation on surface properties of ovalbumin and lysozyme[J].Journal of the Agricultural and Biological Chemistry,1981,45(12):2755-2760.DOI:10.1080/00021369.1981.10864964.
[26]伍林,欧阳兆辉,曹淑超,等.拉曼光谱技术的应用及研究进展[J].光散射学报,2005,17(2):180-186.DOI:10.3969/j.issn.1004-5929.2005.02.013.
[27]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.
[28]CARECHE M,GARCIA M L,HERRERO A,et al.Structural properties of aggregates from frozen stored hake muscle proteins[J].Journal of Food Science,2002,67(8):2827-2832.DOI:10.1111/j.1365-2621.2002.tb08823.x.
[29]HERRERO A M.Raman spectroscopy for monitoring protein structure in muscle food systems[J].Critical Reviews in Food Science and Nutrition,2008,48(6):512-523.DOI:10.1080/10408390701537385.
[30]CHOI S M,MA C Y.Structural characterization of globulin from common buckwheat(Fagopyrum esculentum Moench)using circular dichroism and Raman spectroscopy[J].Food Chemistry,2007,102(1):150-160.DOI:10.1016/j.foodchem.2006.05.011.
[31]胡飞华.梅鱼鱼糜超高压凝胶化工艺及凝胶机理的研究[D].杭州:浙江工商大学,2010:22-40.
[32]ONWULATA C I,ISOBE S,TOMASULA P M,et al.Properties of whey protein isolates extruded under acidic and alkaline conditions[J].Journal of Dairy Science,2006,89(1):71-81.DOI:10.3168/jds.s0022-0302(06)72070-7.
[33]杨华,陆森超,张慧恩,等.超高压处理对养殖大黄鱼风味及品质的影响[J].食品科学,2014,35(16):244-249.DOI:10.7506/spkx1002-6630-201416047.
[34]付强.超高压处理对鲢鱼鱼糜品质特性的影响[D].上海:上海海洋大学,2015:21-80.
[35]仪淑敏,马兴胜,励建荣,等.超高压诱导鱼糜凝胶形成中水分特性及凝胶强度的相关性研究[J].中国食品学报,2015,15(7):26-31.DOI:10.16429/j.1009-7848.2015.07.004.
[36]刘坚,李艳红,缪铭,等.超高压对不同缓冲体系中鹰嘴豆分离蛋白溶解性的影响[J].食品工业科技,2007(11):90-92.DOI:10.13386/j.issn1002-0306.2007.11.017.
[37]王喜波,王健,张泽宇,等.物理改性对大豆蛋白柔性与乳化性的影响及其相关性分析[J].农业机械学报,2017,48(7):339-344.DOI:10.6041/j.issn.1000-1298.2017.07.043.
[38]郑捷,尚校兰,刘安军.超高压处理对海鲈鱼鱼肉凝胶形成作用[J].食品科学,2013,34(19):88-92.DOI:10.7506/spkx1002-6630-201319020.
[39]刘书成,邓倩琳,黄万有,等.超高压处理对凡纳滨对虾虾仁蛋白质和微观结构的影响[J].水产学报,2017,41(6):877-887.DOI:10.11964/jfc.20170110674.
[40]周敏,陆维克,陆金金,等.超高压处理对副溶血性弧菌胞内蛋白质的影响[J].食品科学,2017,38(1):99-104.DOI:10.7506/spkx1002-6630-201701016.
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