低温胁迫肌原纤维蛋白结构和热稳定性的变化
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摘要
本文研究了冻藏温度(-5,-18,-26,-35,-70℃)、冻藏时间(0,1,3,6,9,12 mon)对猪肉背最长肌中的蛋白质结构(ATPase酶活性、巯基含量、SDS-聚丙烯酰氨凝胶电泳)和热稳定性的影响。结果表明:随着冻藏温度的降低,12 mon蛋白质总巯基含量、活性巯基含量、Ca~(2+)-ATPase活性、K~+-ATPase活性呈递减趋势;通过DSC测定蛋白的热稳定性,冻藏过程中肌肉蛋白的热转变温度和焓值均降低,且冻结温度越高冻藏时间越长,温度和焓值降低得越明显;蛋白的SDS-PAGE电泳图谱结果显示,冻藏时间越长冻藏温度越高,蛋白质分子降解越明显。
The effect of freezing temperature(-5,-18,-26,-35 and-70 ℃) and freezing storage time(0, 1,3, 6, 9, 12 mon) on protein structure(ATPase activity, sulfhydryl content, SDS-PAGE) and thermal stability of Myofibrillar protein were studied. The results show that with the decrease of freezing storage temperature, after frozen storage 12 mon, the total sulfhydryl content of MP, the activity sulfhydryl content of MP, the Ca~(2+)-ATPase activity of MP, the K~+-ATPase activity of MP decreased progressively. Myofibrillar protein thermal stability is determination by DSC during frozen storage. The result is: myofibrillar protein thermal transition temperature and enthalpy values are the lower. Myofibrillar protein thermal transition temperature and enthalpy values significantly decreased; the longer freezing time, the higher freezing temperature, myofibrillar protein thermal transition temperature and enthalpy values are the lower. The protein of SDS-PAGE electrophoresis patterns showed that the course of frozen storage occurred to protein degradation and denaturation.
The effect of freezing temperature(-5,-18,-26,-35 and-70 ℃) and freezing storage time(0, 1,3, 6, 9, 12 mon) on protein structure(ATPase activity, sulfhydryl content, SDS-PAGE) and thermal stability of Myofibrillar protein were studied. The results show that with the decrease of freezing storage temperature, after frozen storage 12 mon, the total sulfhydryl content of MP, the activity sulfhydryl content of MP, the Ca~(2+)-ATPase activity of MP, the K~+-ATPase activity of MP decreased progressively. Myofibrillar protein thermal stability is determination by DSC during frozen storage. The result is: myofibrillar protein thermal transition temperature and enthalpy values are the lower. Myofibrillar protein thermal transition temperature and enthalpy values significantly decreased; the longer freezing time, the higher freezing temperature, myofibrillar protein thermal transition temperature and enthalpy values are the lower. The protein of SDS-PAGE electrophoresis patterns showed that the course of frozen storage occurred to protein degradation and denaturation.
引文
[1]丁一,黄莉,夏秀芳,等.冻藏对肌原纤维蛋白结构特性与功能特性的影响[J].包装与食品机械,2013, 31(4):55-59.
[2]赵维高,刘文营,黄丽燕,等.食品加工中蛋白质起泡性的研究[J].农产品加工·学刊, 2012, 298(11):69-72.
[3]XIA X F, KONG B H, JING L, et al. Influence of different thawing methods on physicochemical changes and protein oxidation of porcine longissimus muscle[J]. LWT-Food Science and Technology, 2012, 46(1):280-286.
[4]汪兰,吴文锦,乔宇,等.冻藏条件下魔芋葡甘聚糖降解产物对肌原纤维蛋白结构的影响[J].食品科学, 2015, 36(22):244-249.
[5]BENJAKUL S, SUTTHIPAN N. Muscle changes in hard and soft shell crabs during frozen storage[J].LWT-Food Science and Technology, 2009, 42(3):723-729.
[6]BENJAKUL S, BAUER F. Physicochemical and enzymatic changes of cod muscle proteins subjected to different freeze-thaw cycles[J]. Journal of the Science of Food&Agriculture, 2012, 80(8):1143-1150.
[7]SUBRAMANIAN T A. Effect of processing on bacterial population of cuttle fish and crab and determination of bacterial spoilage and rancidity development on frozen storage[J]. Journal of Food Processing&Preservation, 2013, 31(1):13-31.
[8]TAMMATINNA A, BENJAKUL S, VIDESSANGUAN W, et al. Gelling properties of white shrimp(Penaeus vannamei)meat as influenced by setting condition and microbial transglutaminase[J]. LWT-Food Science and Technology, 2007, 40(9):1489-1497.
[9]崔瑞颖,焦学芹,祖铁红,等.冷冻方式对海湾扇贝闭壳肌大分子物质及组织结构的影响[J].食品科技, 2014, 39(2):134-138.
[10]LIU G, XIONG Y L. Contribution of liqid and protein oxidation to rheological differences between chicken white and red muscle myofibrillar proteins[J]. Journal of Agricultural and Food Chemistry,1996, 44(3):779-784.
[11]WELLS J A, WERBER M M, YOUNT R G. Inactivation of myosin subfragment one by cobalt(Ⅱ)/cobalt(III)phenanthroline complexes 2. Cobalt chelation of two critical SH groups[J]. Bochemistry,1979, 18(22):4800-4805.
[12]KATO N, UCHIYAMA H, TSUKAMOTO S, et al.A biochemical study on fish myofibrillar ATPase[J].Nippon Suisan Gakkaishi, 1977, 43(7):857-867.
[13]DI SIPLICIO P, CHEESEMAN K, SLATER T. The reactivity of the SH group of bovine serum albumin with free radicals[J]. Free Radical Research, 1991,14(4):253-262.
[14]LAEMMLIU K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature, 1970, 227(5259):680-685.
[15]曾名湧.几种主要淡水经济鱼类肌肉蛋白质冻结变性机理的研究[D].青岛:中国海洋大学, 2005.
[16]GONCALVES A A. Development of quality index method(QIM)scheme for spiny lobster(Panulirus argus, Latreille, 1804)stored in ice[J]. Food Control, 2015, 47(1):237-245.
[17]LOMIWES D, FAROUK M M, FROST D A, et al.Small heat shock proteins and toughness in intermediate pHu, beef[J]. Meat Science, 2013, 95(3):472-479.
[18]WAGNER J R, ANON M C. Effect of frozen storage on protein denaturation in bovine muscle[J]. International Journal of Food Science&Technology,1986, 21(5):547-558.
[19]WU G, CLERENS S, FAROUK M M. LC MS/MS identification of large structural proteins from bull muscle and their degradation products during post mortem, storage[J]. Food Chemistry, 2014, 150(4):137-144.
[20]SOMPONGSE W, ITOH Y, OBATAKE A. Role of SHa in the polymerization of myosin heavy chain during ice storage of Carp Actomyosin[J]. Fisheries Science, 1996, 62(1):110-113.
[21]SHIGERU H, SHURYO N. Contribution of Hydrophobicity, Net charge and Sulfhydryl groups to thermal properties of Ovalbumin[J]. Canadian Institute of Food Science&Technology Journal, 1985,18(4):290-295.
[22]HAMRE K,?YVIND LIE, SANDNES K. Development of lipid oxidation and flesh colour in frozen stored fillets of Norwegian spring-spawning herring(Clupea harengus, L.). Effects of treatment with ascorbic acid[J]. Food Chemistry, 2003, 82(3):447-453.
[23]HU Y, JI R, JIANG H, et al. Participation of cathepsin L in modori phenomenon in carp(Cyprinus carpio)surimi gel[J]. Food Chemistry, 2012,134(4):2014-2020.
[24]LIN S B, CHENL C, CHEN H H. The change of thermal gelation properties of horse mackerel mince led by protein denaturation occurring in frozen storage and consequential air floatation wash[J]. Food Research International, 2005, 38(1):19-27.
[25]徐晨晨,罗海玲.钙蛋白酶系统对肌肉系水力的影响研究进展[J].肉类研究, 2015, 29(6):29-32.
[26]THANONKAEW A, BENJAKUL S, VISESSANGUAN W, et al. Gelling properties of white shrimp(Penaeus vannamei)meat as influenced by setting condition and microbial transglutaminase[J]. LWTFood Science and Technology, 2007, 40(9):1489-1497.
[27]FU L, ZHANG S, ZHOU J, et al. Alterations of protein expression in the muscle of pacific white shrimp(Litopenaeus vannamei)contribute to postmortem changes[J]. Journal of Shellfish Research,2014, 33(3):815-823.
[28]FARAH B, NAZLIN K, HOWELl. Changes in the texture and structure of cod and haddock fillet s during frozen storage[J]. Food Hydrocolloids, 2002,16(4):313-319.
[29]BENJENJAKUL S, BAUER F. Physicochmical and enzymatic changes of Cod muscle proteins subjected to different freeze-thaw cycles[J]. Journal of the Science of Food and Agriculture, 2000, 80(8):1143-1150.
[30]DELLES R M, XIONG Y L. The effect of protein oxidation on hydration and water-binding in pork packaged in an oxygen-enriched atmosphere[J]. Meat Science, 2014, 97(2):181-188.
[2]赵维高,刘文营,黄丽燕,等.食品加工中蛋白质起泡性的研究[J].农产品加工·学刊, 2012, 298(11):69-72.
[3]XIA X F, KONG B H, JING L, et al. Influence of different thawing methods on physicochemical changes and protein oxidation of porcine longissimus muscle[J]. LWT-Food Science and Technology, 2012, 46(1):280-286.
[4]汪兰,吴文锦,乔宇,等.冻藏条件下魔芋葡甘聚糖降解产物对肌原纤维蛋白结构的影响[J].食品科学, 2015, 36(22):244-249.
[5]BENJAKUL S, SUTTHIPAN N. Muscle changes in hard and soft shell crabs during frozen storage[J].LWT-Food Science and Technology, 2009, 42(3):723-729.
[6]BENJAKUL S, BAUER F. Physicochemical and enzymatic changes of cod muscle proteins subjected to different freeze-thaw cycles[J]. Journal of the Science of Food&Agriculture, 2012, 80(8):1143-1150.
[7]SUBRAMANIAN T A. Effect of processing on bacterial population of cuttle fish and crab and determination of bacterial spoilage and rancidity development on frozen storage[J]. Journal of Food Processing&Preservation, 2013, 31(1):13-31.
[8]TAMMATINNA A, BENJAKUL S, VIDESSANGUAN W, et al. Gelling properties of white shrimp(Penaeus vannamei)meat as influenced by setting condition and microbial transglutaminase[J]. LWT-Food Science and Technology, 2007, 40(9):1489-1497.
[9]崔瑞颖,焦学芹,祖铁红,等.冷冻方式对海湾扇贝闭壳肌大分子物质及组织结构的影响[J].食品科技, 2014, 39(2):134-138.
[10]LIU G, XIONG Y L. Contribution of liqid and protein oxidation to rheological differences between chicken white and red muscle myofibrillar proteins[J]. Journal of Agricultural and Food Chemistry,1996, 44(3):779-784.
[11]WELLS J A, WERBER M M, YOUNT R G. Inactivation of myosin subfragment one by cobalt(Ⅱ)/cobalt(III)phenanthroline complexes 2. Cobalt chelation of two critical SH groups[J]. Bochemistry,1979, 18(22):4800-4805.
[12]KATO N, UCHIYAMA H, TSUKAMOTO S, et al.A biochemical study on fish myofibrillar ATPase[J].Nippon Suisan Gakkaishi, 1977, 43(7):857-867.
[13]DI SIPLICIO P, CHEESEMAN K, SLATER T. The reactivity of the SH group of bovine serum albumin with free radicals[J]. Free Radical Research, 1991,14(4):253-262.
[14]LAEMMLIU K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature, 1970, 227(5259):680-685.
[15]曾名湧.几种主要淡水经济鱼类肌肉蛋白质冻结变性机理的研究[D].青岛:中国海洋大学, 2005.
[16]GONCALVES A A. Development of quality index method(QIM)scheme for spiny lobster(Panulirus argus, Latreille, 1804)stored in ice[J]. Food Control, 2015, 47(1):237-245.
[17]LOMIWES D, FAROUK M M, FROST D A, et al.Small heat shock proteins and toughness in intermediate pHu, beef[J]. Meat Science, 2013, 95(3):472-479.
[18]WAGNER J R, ANON M C. Effect of frozen storage on protein denaturation in bovine muscle[J]. International Journal of Food Science&Technology,1986, 21(5):547-558.
[19]WU G, CLERENS S, FAROUK M M. LC MS/MS identification of large structural proteins from bull muscle and their degradation products during post mortem, storage[J]. Food Chemistry, 2014, 150(4):137-144.
[20]SOMPONGSE W, ITOH Y, OBATAKE A. Role of SHa in the polymerization of myosin heavy chain during ice storage of Carp Actomyosin[J]. Fisheries Science, 1996, 62(1):110-113.
[21]SHIGERU H, SHURYO N. Contribution of Hydrophobicity, Net charge and Sulfhydryl groups to thermal properties of Ovalbumin[J]. Canadian Institute of Food Science&Technology Journal, 1985,18(4):290-295.
[22]HAMRE K,?YVIND LIE, SANDNES K. Development of lipid oxidation and flesh colour in frozen stored fillets of Norwegian spring-spawning herring(Clupea harengus, L.). Effects of treatment with ascorbic acid[J]. Food Chemistry, 2003, 82(3):447-453.
[23]HU Y, JI R, JIANG H, et al. Participation of cathepsin L in modori phenomenon in carp(Cyprinus carpio)surimi gel[J]. Food Chemistry, 2012,134(4):2014-2020.
[24]LIN S B, CHENL C, CHEN H H. The change of thermal gelation properties of horse mackerel mince led by protein denaturation occurring in frozen storage and consequential air floatation wash[J]. Food Research International, 2005, 38(1):19-27.
[25]徐晨晨,罗海玲.钙蛋白酶系统对肌肉系水力的影响研究进展[J].肉类研究, 2015, 29(6):29-32.
[26]THANONKAEW A, BENJAKUL S, VISESSANGUAN W, et al. Gelling properties of white shrimp(Penaeus vannamei)meat as influenced by setting condition and microbial transglutaminase[J]. LWTFood Science and Technology, 2007, 40(9):1489-1497.
[27]FU L, ZHANG S, ZHOU J, et al. Alterations of protein expression in the muscle of pacific white shrimp(Litopenaeus vannamei)contribute to postmortem changes[J]. Journal of Shellfish Research,2014, 33(3):815-823.
[28]FARAH B, NAZLIN K, HOWELl. Changes in the texture and structure of cod and haddock fillet s during frozen storage[J]. Food Hydrocolloids, 2002,16(4):313-319.
[29]BENJENJAKUL S, BAUER F. Physicochmical and enzymatic changes of Cod muscle proteins subjected to different freeze-thaw cycles[J]. Journal of the Science of Food and Agriculture, 2000, 80(8):1143-1150.
[30]DELLES R M, XIONG Y L. The effect of protein oxidation on hydration and water-binding in pork packaged in an oxygen-enriched atmosphere[J]. Meat Science, 2014, 97(2):181-188.