骨架蛋白降解对冰鲜草鱼质构的影响
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摘要
利用免疫组化技术检测草鱼背肌白肉中骨架蛋白在冰藏条件下的变化,同时通过超微结构、剪切力、滴水损失率分析草鱼背肌组织精微结构和质构特性的变化,以期阐明草鱼骨架蛋白降解与鱼肉质构劣化之间的关系,从而为调控冰鲜淡水鱼品质提供理论依据。结果显示:冰藏21 d内伴肌球蛋白、伴肌动蛋白、抗肌营养不良蛋白分别降解了76.10%、78.21%、71.14%;冰藏14 d后草鱼肌纤维明暗带模糊,Z带、M带被破坏,肌纤维结构出现严重断裂松散现象。冰藏10 d内剪切力由81 N快速下降至32 N,滴水损失率由1.6%增加至7.8%;将骨架蛋白的灰度与剪切力及滴水损失率进行相关性分析发现,剪切力与伴肌球蛋白、抗肌营养不良蛋白的灰度呈极显著正相关(P<0.01);滴水损失率与伴肌球蛋白、抗肌营养不良蛋白的灰度值呈极显著负相关(P<0.01)。综上所述,骨架蛋白降解可能是草鱼在冰藏期间肌纤维结构破坏以及质构劣化的重要原因。
The aim of this work was to investigate the changes of cytoskeletal proteins in the dorsal muscle of grass carp(Ctenopharyngodon idella) during ice storage by immunohistochemistry,and to analyze the changes in the microstructure and textural properties by ultrastructural observation and measurement of shear force and drip loss.In addition,this study clarified the relationship between the degradation of cytoskeleton protein and the deterioration of fish texture,which will a provide theoretical foundation for the quality preservation of freshwater fish.The results showed that cytoskeleton proteins significantly degraded during ice storage.Titin,nebulin,and dystrophin degraded by 76.10%,78.21%,and 71.14%,respectively.The light and dark bands of grass carp muscle fibers were unclear after 14 days of ice storage.The Z-band and M-band were degraded.and the muscle fiber structure severely broke or became loose.Meanwhile,shear force rapidly decreased from 81 N to 32 N within 10 days of storage,while drop loss increased from 1.6% to 7.8%.Correlation analysis indicated a significantly positive correlation between shear force and the integrated option density(IOD) of titin and dystrophin(P<0.01).In contrast,a significantly negative correlation was observed between drip loss and the IOD of titin and dystrophin(P<0.01).The above results showed that the degradation of cytoskeletal proteins may be an important cause of the destruction of muscle fiber and textural deterioration of grass carp during ice storage.
The aim of this work was to investigate the changes of cytoskeletal proteins in the dorsal muscle of grass carp(Ctenopharyngodon idella) during ice storage by immunohistochemistry,and to analyze the changes in the microstructure and textural properties by ultrastructural observation and measurement of shear force and drip loss.In addition,this study clarified the relationship between the degradation of cytoskeleton protein and the deterioration of fish texture,which will a provide theoretical foundation for the quality preservation of freshwater fish.The results showed that cytoskeleton proteins significantly degraded during ice storage.Titin,nebulin,and dystrophin degraded by 76.10%,78.21%,and 71.14%,respectively.The light and dark bands of grass carp muscle fibers were unclear after 14 days of ice storage.The Z-band and M-band were degraded.and the muscle fiber structure severely broke or became loose.Meanwhile,shear force rapidly decreased from 81 N to 32 N within 10 days of storage,while drop loss increased from 1.6% to 7.8%.Correlation analysis indicated a significantly positive correlation between shear force and the integrated option density(IOD) of titin and dystrophin(P<0.01).In contrast,a significantly negative correlation was observed between drip loss and the IOD of titin and dystrophin(P<0.01).The above results showed that the degradation of cytoskeletal proteins may be an important cause of the destruction of muscle fiber and textural deterioration of grass carp during ice storage.
引文
[1]农业部渔业渔政管理局.2017中国渔业统计年鉴[M].北京:中国农业出版社,2017:17-25.
[2]李倩,张龙腾,罗永康.主要淡水鱼类价格与营养成分、感官评价的相关性研究[J].科学养鱼,2016(7):77-78.DOI:10.14184/j.cnki.issn1004-843x.2016.07.048.
[3]欧阳芳芳,王建辉,陈奇,等.草鱼贮藏期间肌肉AT P关联物及K值的动态变化[J].食品与机械,2016,32(3):137-140;159.DOI:10.13652/j.issn.1003-5788.2016.03.030.
[4]KAALE L D,EIKEVIK T M.A study of the ice crystal sizes of red muscle of pre-rigor Atlantic salmon(Salmo salar)fillets during superchilled storage[J].Journal of Food Engineering,2013,119(3):544-551.DOI:10.1016/j.jfoodeng.2013.06.002.
[5]FENG X,BANSAL N,YANG H S.Fish gelatin combined with chitosan coating inhibits myofibril degradation of golden pomfret(Trachinotus blochii)fillet during cold storage[J].Food Chemistry,2016,200:283-292.DOI:10.1016/j.f00dchem.2016.01.030.
[6]邹聪聪,卢蓓,张石天,等.快速冷冻对大黄鱼、鲳鱼肌原纤维超微结构的影响[J].水产科技情报,2017,44(5):276-279;281.DOI:10.16446/j.cnki.1001-1994.2017.05.012.
[7]曹雷鹏,牛丽红,孙晓华,等.草鱼冰藏期间新鲜度及热加工性质的变化[J].食品科学,2016,37(12):267-272.DOI:10.7506/spkx1002-6630-201612048.
[8]GE L H,XU Y S,JIANG X Q,et al.Broad-spectrum inhibition of proteolytic enzymes by allicin and application in mitigating textural deterioration of ice-stored grass carp(Ctenopharyngodon idella)fillets[J].International Journal of Food Science&Technology,2016,51(4):902-910.DOI:10.1111/ijfs.13047.
[9]NEMOVA N N,LYSENKO L A,KANTSEROVA N P.Degradation of skeletal muscle protein during growth and development of salmonid fish[J].Russian Journal of Developmental Biology,2016,47(4):161-172.DOI:10.1134/S1062360416040068.
[10]HUANG J C,YANG J,HUANG F,et al.Effect of fast pH decline during the early postmortem period on calpain activity and cytoskeletal protein degradation of broiler M.pectoralis major[J].Poultry Science,2016,95(10):2455-2463.DOI:10.3382/ps/pew206.
[11]HUANG F,HUANG M,ZHOU G,et al.In vitro proteolysis of myofibrillar proteins from beef skeletal muscle by caspase-3 and caspase-6[J].Journal of Agricultural Food Chemistry,2011,59(17):9658-9663.DOI:10.1021/jf202129.
[12]LAITILA J,HANIF M,PAETAU A,et al.Expression of multiple nebulin isoforms in human skeletal muscle and brain[J].Muscle&Nerve,2012,46(5):730-734.DOI:10.1002/mus.23380.
[13]ROYUELA M,FRAILE B,DEMIGUELM P,et al.Immunohistochemical study and western blotting analysis of titinlike proteins in the striated muscle of drosophila melanogaster and in the striated and smooth muscle of the oligochaete eisenia foetida[J].Microscopy Research&Technique,2015,35(4):349-356.
[14]CRUZ-RICO G,AVILéS-SALAS A,SEGURA-GONZáLEZ M,et al.Diagnosis of eml4-alk translocation with fish,immunohistochemistry,and real-time polymerase chain reaction in patients with non-small cell lung cancer[J].American Journal of Clinical Oncology,2017,40(6):631-638.
[15]ULLAH S,LE Y,BIAN X G,et al.Fine structure of zebrafish(Danio rerio)spermatozoa[J].Pakistan Veterinary Journal,2014,8(4):253-318.
[16]ZANG Jinhong,XU Yanshun,XIA Wenshui,et al.The impact of desmin on texture and water-holding capacity of ice-stored grass carp(Ctenopharyngodon idella)fillet[J].International Journal of Food Science&Technology,2017,52(2):464-471.DOI:10.1111/ijfs.13302.
[17]MU Gang,BERGSSON A B,JONSSON A,et al.The effects of ambient temperature and holding time during processing on drip of saithe(Pollachius virens)and deepwater redfish(Sebastes mentella)fillets[J].Journal of Food Engineering,2017,200:40-46.DOI:10.1016/j.jfoodeng.2016.12.019.
[18]LABEIT S,KOLMERER B.Titins:giant proteins in charge of muscle ultrastructure and elasticity[J].Science,1995,270:293-296.DOI:10.1126/science.270.5234.293.
[19]LI Y P,LIU R,ZHANG W G,et al.Effect of nitric oxide onμ-calpain activation,protein proteolysis,and protein oxidation of pork during post-mortem aging[J].Journal of Agricultural Food Chemisty,2014,62(25):5972-5977.DOI:10.1021/jf501332d.
[20]ANDERSON T J,PARRISH F C.Postmortem degradation of titin and nebulin of beef steaks varying in tenderness[J].Journal of Food Science,1989,54(3):748-749.DOI:10.1111/j.1365-2621.1989.tb04695.x.
[21]MCELHINNY A S,KAZMIERSKI S T,LABEIT S,et al.Nebulin:the nebulous,multifunctional giant of striated muscle[J].Trends in Cardiovascular Medicine,2003,13(5):195-201.DOI:10.1016/S1050-1738(03)00076-8.
[22]CUI B,ZHANG Y,WANG F,et al.Technical note:the effect of carcass deboning technique on the meat quality of cattle 1[J].Journal of Animal Science,2016,94(10):4427-4433.DOI:10.2527/jas.2016-0550.
[23]HUANG M,HUANG F,XUE M,et al.The effect of active caspase-3on degradation of chicken myofibrillar proteins and structure of myofibrils[J].Food Chemistry,2011,128(1):22-27.DOI:10.1016/j.foodchem.2011.02.062.
[24]杞斯卡娜,郑伟,陈韬,等.骨架蛋白肌联蛋白、伴肌动蛋白、纽蛋白的降解对冷却猪肉持水性的影响[J].肉类研究,2013,27(3):42-45.
[25]SKUK D,TREMBLAY J P.Confirmation of donor-derived dystrophin in a duchenne muscular dystrophy patient allotransplanted with normal myoblasts[J].Muscle&Nerve,2016,54(5):979-981.DOI:10.1002/mus.25129.
[26]李经伦,吕英姿,何晓英,等.免疫组化联合放射自显影术观察骨髓间充质干细胞在肌营养不良鼠表达抗肌萎缩蛋白[J].泸州医学院学报,2013,36(4):320-324.
[27]宋楠,杨晓凤.假肥大型肌营养不良肌细胞抗肌萎缩蛋白的免疫组化研究[J].中国组织化学与细胞化学杂志,2010,19(4):377-379.
[28]BASS A H,MARCHATERRE M A.Sound-generating(sonic)motor system in a teleost fish(Porichthys notatus):sexual polymorphism in the ultrastructure of myofibrils[J].Journal of Comparative Neurology,1989,286(2):d141-d153.DOI:10.1002/cne.902860202.
[29]KIMURA S,MATSUURA T,OHTSUKA S,et al.Characterization and localization ofα-connectin(titin 1):an elastic protein isolated from rabbit skeletal muscle[J].Journal of Muscle Research Cell Motility,1992,13(1):39-47.
[30]TAKAHASHI K,NAKAMURA F,INOUE A.Postmortem changes in the actin-myosin interaction of rabbit skeletal muscle[J].Journal of Biochemistry,1981,89(1):321-324.DOI:10.1093/oxfordjournals.jbchem.a133197.
[31]黄鸿兵,王道营,张牧晗,等.草鱼冷藏过程中肌原纤维超微结构及质构的变化[J].天津农业科学,2015,21(12):58-60.
[32]CHENG J H,SUN D W,HAN Z,et al.Texture and structure measurements and analyses for evaluation of fish and fillet freshness quality:a review[J].Comprehensive Reviews in Food Science and Food Safety,2013,13(1):52-61.DOI:10.1111/1541-4337.12043.
[33]MOHAMED B A,ELLIS N,KIM C S,et al.Engineered biochar from microwave-assisted catalytic pyrolysis of switchgrass for increasing water-holding capacity and fertility of sandy soil[J].Science of the Total Environment,2016,93(5):387-397.DOI:10.1016/j.scitotenv.2016.04.169.
[34]THOMAS A R,GONDOZA H,HOFFMAN L C,et al.The roles of the proteasome,and cathepsins B,L,H and D,in ostrich meat tenderisation[J].Meat Science,2004,67(1):113-120.DOI:10.1016/j.meatsci.2003.10.001.
[35]BARBUT S,SOSNICKI A A,LONERGAN S M,et al.Progress in reducing the pale,soft and exudative(PSE)problem in pork and poultry meat[J].Meat Science,2008,79(1):46-63.
[36]DELBARRE-LADRAT C,CHéRET R,TAYLOR R,et al.Trends in postmortem aging in fish:understanding of proteolysis and disorganization of the myofibrillar structure[J].Critical Reviews in Food Science and Nutrition,2006,46(5):409-421.
[37]REES M P,TROUT G R,WARNER R D.The influence of the rate of pH decline on the rate of ageing for pork.I:interaction with method of suspension[J].Meat Science,2003,65(2):791-804.DOI:10.1016/S0309-1740(02)00285-1.
[38]AYALA M D,GARCíA-ALCáZAR A,ABDEL I,et al.Effect of thermal treatment on muscle tissue structure and ultrastructure of wild and farmed sea bass,Dicentrarchus labrax L[J].Aquaculture International,2010,18(6):1137-1149.
[39]SCH?FER A,ROSENVOLD K,PURSLOW P P,et al.Physiological and structural events post mortem of importance for drip loss in pork[J].Meat Science,2002,61(4):355-366.
[40]吴霜,陈韬,刘劭,等.猪宰后正常肉与PSE肉中肌联蛋白和伴肌动蛋白变化[J].食品工业,2015,36(10):187-192.
[2]李倩,张龙腾,罗永康.主要淡水鱼类价格与营养成分、感官评价的相关性研究[J].科学养鱼,2016(7):77-78.DOI:10.14184/j.cnki.issn1004-843x.2016.07.048.
[3]欧阳芳芳,王建辉,陈奇,等.草鱼贮藏期间肌肉AT P关联物及K值的动态变化[J].食品与机械,2016,32(3):137-140;159.DOI:10.13652/j.issn.1003-5788.2016.03.030.
[4]KAALE L D,EIKEVIK T M.A study of the ice crystal sizes of red muscle of pre-rigor Atlantic salmon(Salmo salar)fillets during superchilled storage[J].Journal of Food Engineering,2013,119(3):544-551.DOI:10.1016/j.jfoodeng.2013.06.002.
[5]FENG X,BANSAL N,YANG H S.Fish gelatin combined with chitosan coating inhibits myofibril degradation of golden pomfret(Trachinotus blochii)fillet during cold storage[J].Food Chemistry,2016,200:283-292.DOI:10.1016/j.f00dchem.2016.01.030.
[6]邹聪聪,卢蓓,张石天,等.快速冷冻对大黄鱼、鲳鱼肌原纤维超微结构的影响[J].水产科技情报,2017,44(5):276-279;281.DOI:10.16446/j.cnki.1001-1994.2017.05.012.
[7]曹雷鹏,牛丽红,孙晓华,等.草鱼冰藏期间新鲜度及热加工性质的变化[J].食品科学,2016,37(12):267-272.DOI:10.7506/spkx1002-6630-201612048.
[8]GE L H,XU Y S,JIANG X Q,et al.Broad-spectrum inhibition of proteolytic enzymes by allicin and application in mitigating textural deterioration of ice-stored grass carp(Ctenopharyngodon idella)fillets[J].International Journal of Food Science&Technology,2016,51(4):902-910.DOI:10.1111/ijfs.13047.
[9]NEMOVA N N,LYSENKO L A,KANTSEROVA N P.Degradation of skeletal muscle protein during growth and development of salmonid fish[J].Russian Journal of Developmental Biology,2016,47(4):161-172.DOI:10.1134/S1062360416040068.
[10]HUANG J C,YANG J,HUANG F,et al.Effect of fast pH decline during the early postmortem period on calpain activity and cytoskeletal protein degradation of broiler M.pectoralis major[J].Poultry Science,2016,95(10):2455-2463.DOI:10.3382/ps/pew206.
[11]HUANG F,HUANG M,ZHOU G,et al.In vitro proteolysis of myofibrillar proteins from beef skeletal muscle by caspase-3 and caspase-6[J].Journal of Agricultural Food Chemistry,2011,59(17):9658-9663.DOI:10.1021/jf202129.
[12]LAITILA J,HANIF M,PAETAU A,et al.Expression of multiple nebulin isoforms in human skeletal muscle and brain[J].Muscle&Nerve,2012,46(5):730-734.DOI:10.1002/mus.23380.
[13]ROYUELA M,FRAILE B,DEMIGUELM P,et al.Immunohistochemical study and western blotting analysis of titinlike proteins in the striated muscle of drosophila melanogaster and in the striated and smooth muscle of the oligochaete eisenia foetida[J].Microscopy Research&Technique,2015,35(4):349-356.
[14]CRUZ-RICO G,AVILéS-SALAS A,SEGURA-GONZáLEZ M,et al.Diagnosis of eml4-alk translocation with fish,immunohistochemistry,and real-time polymerase chain reaction in patients with non-small cell lung cancer[J].American Journal of Clinical Oncology,2017,40(6):631-638.
[15]ULLAH S,LE Y,BIAN X G,et al.Fine structure of zebrafish(Danio rerio)spermatozoa[J].Pakistan Veterinary Journal,2014,8(4):253-318.
[16]ZANG Jinhong,XU Yanshun,XIA Wenshui,et al.The impact of desmin on texture and water-holding capacity of ice-stored grass carp(Ctenopharyngodon idella)fillet[J].International Journal of Food Science&Technology,2017,52(2):464-471.DOI:10.1111/ijfs.13302.
[17]MU Gang,BERGSSON A B,JONSSON A,et al.The effects of ambient temperature and holding time during processing on drip of saithe(Pollachius virens)and deepwater redfish(Sebastes mentella)fillets[J].Journal of Food Engineering,2017,200:40-46.DOI:10.1016/j.jfoodeng.2016.12.019.
[18]LABEIT S,KOLMERER B.Titins:giant proteins in charge of muscle ultrastructure and elasticity[J].Science,1995,270:293-296.DOI:10.1126/science.270.5234.293.
[19]LI Y P,LIU R,ZHANG W G,et al.Effect of nitric oxide onμ-calpain activation,protein proteolysis,and protein oxidation of pork during post-mortem aging[J].Journal of Agricultural Food Chemisty,2014,62(25):5972-5977.DOI:10.1021/jf501332d.
[20]ANDERSON T J,PARRISH F C.Postmortem degradation of titin and nebulin of beef steaks varying in tenderness[J].Journal of Food Science,1989,54(3):748-749.DOI:10.1111/j.1365-2621.1989.tb04695.x.
[21]MCELHINNY A S,KAZMIERSKI S T,LABEIT S,et al.Nebulin:the nebulous,multifunctional giant of striated muscle[J].Trends in Cardiovascular Medicine,2003,13(5):195-201.DOI:10.1016/S1050-1738(03)00076-8.
[22]CUI B,ZHANG Y,WANG F,et al.Technical note:the effect of carcass deboning technique on the meat quality of cattle 1[J].Journal of Animal Science,2016,94(10):4427-4433.DOI:10.2527/jas.2016-0550.
[23]HUANG M,HUANG F,XUE M,et al.The effect of active caspase-3on degradation of chicken myofibrillar proteins and structure of myofibrils[J].Food Chemistry,2011,128(1):22-27.DOI:10.1016/j.foodchem.2011.02.062.
[24]杞斯卡娜,郑伟,陈韬,等.骨架蛋白肌联蛋白、伴肌动蛋白、纽蛋白的降解对冷却猪肉持水性的影响[J].肉类研究,2013,27(3):42-45.
[25]SKUK D,TREMBLAY J P.Confirmation of donor-derived dystrophin in a duchenne muscular dystrophy patient allotransplanted with normal myoblasts[J].Muscle&Nerve,2016,54(5):979-981.DOI:10.1002/mus.25129.
[26]李经伦,吕英姿,何晓英,等.免疫组化联合放射自显影术观察骨髓间充质干细胞在肌营养不良鼠表达抗肌萎缩蛋白[J].泸州医学院学报,2013,36(4):320-324.
[27]宋楠,杨晓凤.假肥大型肌营养不良肌细胞抗肌萎缩蛋白的免疫组化研究[J].中国组织化学与细胞化学杂志,2010,19(4):377-379.
[28]BASS A H,MARCHATERRE M A.Sound-generating(sonic)motor system in a teleost fish(Porichthys notatus):sexual polymorphism in the ultrastructure of myofibrils[J].Journal of Comparative Neurology,1989,286(2):d141-d153.DOI:10.1002/cne.902860202.
[29]KIMURA S,MATSUURA T,OHTSUKA S,et al.Characterization and localization ofα-connectin(titin 1):an elastic protein isolated from rabbit skeletal muscle[J].Journal of Muscle Research Cell Motility,1992,13(1):39-47.
[30]TAKAHASHI K,NAKAMURA F,INOUE A.Postmortem changes in the actin-myosin interaction of rabbit skeletal muscle[J].Journal of Biochemistry,1981,89(1):321-324.DOI:10.1093/oxfordjournals.jbchem.a133197.
[31]黄鸿兵,王道营,张牧晗,等.草鱼冷藏过程中肌原纤维超微结构及质构的变化[J].天津农业科学,2015,21(12):58-60.
[32]CHENG J H,SUN D W,HAN Z,et al.Texture and structure measurements and analyses for evaluation of fish and fillet freshness quality:a review[J].Comprehensive Reviews in Food Science and Food Safety,2013,13(1):52-61.DOI:10.1111/1541-4337.12043.
[33]MOHAMED B A,ELLIS N,KIM C S,et al.Engineered biochar from microwave-assisted catalytic pyrolysis of switchgrass for increasing water-holding capacity and fertility of sandy soil[J].Science of the Total Environment,2016,93(5):387-397.DOI:10.1016/j.scitotenv.2016.04.169.
[34]THOMAS A R,GONDOZA H,HOFFMAN L C,et al.The roles of the proteasome,and cathepsins B,L,H and D,in ostrich meat tenderisation[J].Meat Science,2004,67(1):113-120.DOI:10.1016/j.meatsci.2003.10.001.
[35]BARBUT S,SOSNICKI A A,LONERGAN S M,et al.Progress in reducing the pale,soft and exudative(PSE)problem in pork and poultry meat[J].Meat Science,2008,79(1):46-63.
[36]DELBARRE-LADRAT C,CHéRET R,TAYLOR R,et al.Trends in postmortem aging in fish:understanding of proteolysis and disorganization of the myofibrillar structure[J].Critical Reviews in Food Science and Nutrition,2006,46(5):409-421.
[37]REES M P,TROUT G R,WARNER R D.The influence of the rate of pH decline on the rate of ageing for pork.I:interaction with method of suspension[J].Meat Science,2003,65(2):791-804.DOI:10.1016/S0309-1740(02)00285-1.
[38]AYALA M D,GARCíA-ALCáZAR A,ABDEL I,et al.Effect of thermal treatment on muscle tissue structure and ultrastructure of wild and farmed sea bass,Dicentrarchus labrax L[J].Aquaculture International,2010,18(6):1137-1149.
[39]SCH?FER A,ROSENVOLD K,PURSLOW P P,et al.Physiological and structural events post mortem of importance for drip loss in pork[J].Meat Science,2002,61(4):355-366.
[40]吴霜,陈韬,刘劭,等.猪宰后正常肉与PSE肉中肌联蛋白和伴肌动蛋白变化[J].食品工业,2015,36(10):187-192.