壳聚糖复合保鲜对草鱼肌肉蛋白质变化的影响
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摘要
肌肉蛋白质的理化变化是导致淡水鱼贮藏过程中品质下降的重要原因之一,为明确保鲜处理对草鱼肌肉蛋白质变化的影响,本实验将草鱼片分别以壳聚糖复合保鲜液(10 mg/mL壳聚糖+5 mg/mL茶多酚+2 000 U/mL溶菌酶)和无菌水(对照组)处理,通过对比分析草鱼冷藏过程中挥发性盐基氮(total volatile basic nitrogen,TVB-N)含量、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electropheresis,SDS-PAGE)图谱、分子质量分布、活性巯基含量及表面疏水性等蛋白质相关指标的变化,探讨壳聚糖复合保鲜处理对延缓草鱼肌肉蛋白质变化的作用。结果表明:冷藏过程中保鲜组草鱼各项蛋白质相关指标的变化均明显小于对照组,第6～9天差别尤为明显;保鲜处理能抑制TVB-N含量上升、活性巯基含量下降及表面疏水性的变化,表明保鲜组蛋白质分解、氧化、变性程度较低;保鲜组蛋白质的SDS-PAGE和高效液相体积排阻色谱的谱图变化较慢,也证实了保鲜处理在对抑制蛋白质的降解和结构性质变化方面的作用。上述结果表明壳聚糖复合保鲜能一定程度上维持肌肉蛋白质的结构和性质的稳定,从而延缓冷藏过程中草鱼肌肉质量的下降。
Changes in the physicochemical properties of muscle proteins are among the leading causes of quality deterioration during the storage of freshwater fish. The effect of preservative treatment on changes in the physicochemical properties of muscle proteins from freshwater fish is not yet well understood nowadays. Therefore, in this study, two groups of grass carpfillets treated separately with a natural preservative combination of 10 mg/m L chitosan + 5 mg/m L tea polyphenol + 2 000 U/m L lysozyme and sterile water(control group) were analyzed and compared for changes in total volatile basic nitrogen(total volatile basic nitrogen(TVB-N) content, sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) pattern,molecular mass distribution, active sulfhydryl group content and surface hydrophobicity, with the aim of evaluating the effectiveness of the mixed preservative for preventing changes in fish muscle proteins. The results showed that the preservative treatment group exhibited lesser changes in any of the protein properties than did the control group, especially from Day 6 to 9 of cold storage. The preservative treatment prevented the increase in TVB-N content and the decrease in active sulfhydryl group content as well as the change in surface hydrophobicity, indicating reduced protein breakdown,oxidation and denaturation. The slower change in SDS-PAGE and size exclusion-high-performance liquid chromatography profiles provided further evidence supporting the role of the preservative treatment in inhibiting protein degradation and structural change. These results provided evidence that the mixed preservative can stabilize the structure and properties of grass fish muscle proteins to a certain extent and consequently prevent quality deterioration.
Changes in the physicochemical properties of muscle proteins are among the leading causes of quality deterioration during the storage of freshwater fish. The effect of preservative treatment on changes in the physicochemical properties of muscle proteins from freshwater fish is not yet well understood nowadays. Therefore, in this study, two groups of grass carpfillets treated separately with a natural preservative combination of 10 mg/m L chitosan + 5 mg/m L tea polyphenol + 2 000 U/m L lysozyme and sterile water(control group) were analyzed and compared for changes in total volatile basic nitrogen(total volatile basic nitrogen(TVB-N) content, sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) pattern,molecular mass distribution, active sulfhydryl group content and surface hydrophobicity, with the aim of evaluating the effectiveness of the mixed preservative for preventing changes in fish muscle proteins. The results showed that the preservative treatment group exhibited lesser changes in any of the protein properties than did the control group, especially from Day 6 to 9 of cold storage. The preservative treatment prevented the increase in TVB-N content and the decrease in active sulfhydryl group content as well as the change in surface hydrophobicity, indicating reduced protein breakdown,oxidation and denaturation. The slower change in SDS-PAGE and size exclusion-high-performance liquid chromatography profiles provided further evidence supporting the role of the preservative treatment in inhibiting protein degradation and structural change. These results provided evidence that the mixed preservative can stabilize the structure and properties of grass fish muscle proteins to a certain extent and consequently prevent quality deterioration.
引文
[1]农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会.2018中国渔业统计年鉴[M].北京:中国农业出版社,2018:17-38.
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[4]CHOULJENKO A,CHOTIKO A,REYES V,et al.Application of water-soluble chitosan to shrimp for quality retention[J].LWT-Food Science and Technology,2016,74:571-579.DOI:10.1016/j.lwt.2016.08.024.
[5]欧阳涛,赵利,苏伟,等.茶多酚对冷藏草鱼片保鲜效果的研究[J].食品科技,2011,36(9):157-160.DOI:10.13684/j.cnki.spkj.2011.09.013.
[6]张玲,马月,罗永康,等.鲢鱼皮改性明胶复合多肽锌对草鱼的保鲜作用[J].食品科学,2016,37(14):231-236.DOI:10.7506/spkx1002-6630-201614042.
[7]苏辉,谢晶.生物保鲜剂在水产品保鲜中的应用研究进展[J].食品与机械,2013,29(5):265-269.DOI:10.3969/j.issn.1003-5788.2013.05.070.
[8]李应洪,李忠海,付湘晋,等.樟树叶多酚抑菌活性及在冷藏草鱼肉保鲜中的应用[J].食品与机械,2018,34(5):128-130;141.DOI:10.13652/j.issn.1003-5788.2018.05.027.
[9]?ZOGUL Y,?ZYURT G,?ZOGUL F,et al.Freshness assessment of European eel(Anguilla anguilla)by sensory,chemical and microbiological methods[J].Food Chemistry,2005,92:745-751.DOI:10.1016/j.foodchem.2004.08.035.
[10]王满生.多靶栅栏技术在草鱼贮藏过程中对微生物腐败的控制研究[D].长沙:长沙理工大学,2012:40-64.
[11]ADDIS M F,CAPPUCCINELLI R,TEDDE V,et al.Proteomic analysis of muscle tissue from gilthead sea bream(Sparus aurata L.)farmed in offshore floating cages[J].Aquaculture,2010,309(1/2/3/4):245-252.DOI:10.1016/j.aquaculture.2010.08.022.
[12]JIA X H,EKMAN M,GROVE H,et al.Proteome changes in bovine longissimus thoracis muscle during the early postmortem storage period[J].Journal of Proteome Research,2007,6(7):2720-2731.DOI:10.1021/pr070173o.
[13]李强,刘永乐,王发祥,等.草鱼肌肉蛋白质在冷藏过程中的变化[J].食品科学,2013,34(3):55-58.
[14]LIU Yongle,LI Xianghong,ZHOU Xiaoling,et al.Effects of glutaminase deamidation on the structure and solubility of rice glutelin[J].LWT-Food Science and Technology,2011,44(10):2205-2210.DOI:10.1016/j.lwt.2011.05.011.
[15]曾茂茂,王霄,陈洁.蛋白质疏水性测定方法的相关性及适用性[J].食品科学,2011,32(15):117-120.
[16]姜杨,李婷婷,晋高伟,等.草鱼冷藏过程中新鲜度的综合评价[J].食品科学,2014,35(20):281-285.DOI:10.7506/spkx1002-6630-201420055.
[17]RIEBROY S,BENJAKUL S,VISESSANGUAN W,et al.Effect of iced storage of bigeye snapper(Priacanthus tayenus)on the chemical composition,properties and acceptability of Som-fug,a fermented Thai fish mince[J].Food Chemistry,2007,102(1):270-280.DOI:10.1016/j.foodchem.2006.05.017.
[18]李凯风,罗永康,冯启超,等.鱼鳞蛋白酶解物为基料的涂膜剂对鲫的保鲜效果[J].水产学报,2011,35(7):1113-1119.DOI:10.3724/SP.J.1231.2011.17339.
[19]杨胜平,谢晶,佟懿.不同浓度壳聚糖对带鱼冷藏保鲜效果的研究[J].山西农业科学,2010,38(2):77-80.DOI:I0.3969/j.issn.1002-2481.2010.02.22.
[20]老莹,胡文忠,冯可,等.天然抑菌剂的抑菌机理及其在果蔬保鲜中的应用[J].食品与发酵工业,2018,44(9):288-293.DOI:10.13995/j.cnki.11-1802/ts.016977.
[21]梁丽媚,李思东,李程鹏,等.壳聚糖及其衍生物抗菌活性的研究进展[J].广州化工,2017,45(20):6-9;15.
[22]FAN W J,CHI Y L,ZHANG S.The use of a tea polyphenol dip to extend the shelf life of silver carp(Hypophthalmicthys molitrix)during storage in ice[J].Food Chemistry,2008,108(1):148-153.
[23]CALLEWAERT L,MICHIELS C W.Lysozymes in the animal kingdom[J].Journal of Biosciences,2010,35(1):127-160.DOI:10.1007/s12038-010-0015-5.
[24]郑红,苏现波,马良,等.货架期冷藏过程中鳝鱼肉理化指标及蛋白质变化规律[J].食品科学,2018,39(7):215-220.DOI:10.7506/spkx1002-6630-201807032.
[25]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.DOI:10.1080/10408390591000929.
[26]YU Dawei,REGENSTEIN J M,ZANG Jinhong,et al.Inhibitory effects of chitosan-based coatings on endogenous enzyme activities,proteolytic degradation and texture softening of grass carp(Ctenopharyngodon idellus)fillets stored at 4℃[J].Food Chemistry,2018,262:1-6.DOI:10.1016/j.foodchem.2018.04.070.
[27]田悦,杜军保.二硫键和巯基在蛋白质结构功能中的作用及分析方法[J].实用儿科临床杂志,2007(19):1499-1501.
[28]BENJAKUL S,VISESSANGUAN W,THONGKAEW C,et al.Comparative study on physicochemical changes of muscle proteins from some tropical fish during frozen storage[J].Food Research International,2003,36(8):787-795.DOI:10.1016/S0963-9969(03)00073-5.
[29]LU Han,WANG Hang,LUO Yongkang.Changes in protein oxidation,water-holding capacity,and texture of bighead carp fillets under chilled and partial frozen storage[J].Journal of Aquatic Food Product Technology,2017,26(5):566-577.DOI:10.1080/10498850.2016.1238860.
[30]RAMIREZ J A,MARTIN-POLO M O,BANDMAN E.Fish myosin aggregation as affected by freezing and initial physical state[J].Journal of Food Science,2000,65(4):556-560.
[31]刘嘉莉,蓝蔚青,刘大勇,等.壳聚糖在水产品保鲜中应用研究进展[J].食品与机械,2019,35(3):231-236.DOI:10.13652/j.issn.1003-5788.2019.03.041.
[32]NAKAI S.Structure-function relationships of food proteins:with an emphasis on the importance of protein hydrophobicity[J].Journal of Agricultural and Food Chemistry,1983,31(4):676-683.
[33]吕卫金,赵进,汪金林,等.茶多酚延缓冷藏大黄鱼肌原纤维蛋白变性降解机理研究[J].中国食品学报,2014,14(1):60-67.DOI:10.16429/j.1009-7848.2014.01.017.
[34]李向红,王发祥,张付兰,等.冷藏过程中草鱼肌原纤维蛋白结构的变化[J].食品与发酵工业,2015,41(10):177-181.DOI:10.13995/j.cnki.11-1802/ts.201510033.
[35]BENJAKUL S,SEYMOUR T A,MORRISSEY M T,et al.Physicochemical changes in Pacific whiting muscle proteins during iced storage[J].Journal of Food Science,1997,62(4):729-733.
[2]王发祥,王满生,刘永乐,等.低温贮藏下草鱼肉优势腐败菌鉴定及其消长规律[J].食品与发酵工业,2012,2:66-68.DOI:10.13995/j.cnki.11-1802/ts.2012.02.039.
[3]曾名勇,黄海,李八方.鳙肌肉蛋白质生化特性在冻藏过程中的变化[J].水产学报,2003(5):480-485.
[4]CHOULJENKO A,CHOTIKO A,REYES V,et al.Application of water-soluble chitosan to shrimp for quality retention[J].LWT-Food Science and Technology,2016,74:571-579.DOI:10.1016/j.lwt.2016.08.024.
[5]欧阳涛,赵利,苏伟,等.茶多酚对冷藏草鱼片保鲜效果的研究[J].食品科技,2011,36(9):157-160.DOI:10.13684/j.cnki.spkj.2011.09.013.
[6]张玲,马月,罗永康,等.鲢鱼皮改性明胶复合多肽锌对草鱼的保鲜作用[J].食品科学,2016,37(14):231-236.DOI:10.7506/spkx1002-6630-201614042.
[7]苏辉,谢晶.生物保鲜剂在水产品保鲜中的应用研究进展[J].食品与机械,2013,29(5):265-269.DOI:10.3969/j.issn.1003-5788.2013.05.070.
[8]李应洪,李忠海,付湘晋,等.樟树叶多酚抑菌活性及在冷藏草鱼肉保鲜中的应用[J].食品与机械,2018,34(5):128-130;141.DOI:10.13652/j.issn.1003-5788.2018.05.027.
[9]?ZOGUL Y,?ZYURT G,?ZOGUL F,et al.Freshness assessment of European eel(Anguilla anguilla)by sensory,chemical and microbiological methods[J].Food Chemistry,2005,92:745-751.DOI:10.1016/j.foodchem.2004.08.035.
[10]王满生.多靶栅栏技术在草鱼贮藏过程中对微生物腐败的控制研究[D].长沙:长沙理工大学,2012:40-64.
[11]ADDIS M F,CAPPUCCINELLI R,TEDDE V,et al.Proteomic analysis of muscle tissue from gilthead sea bream(Sparus aurata L.)farmed in offshore floating cages[J].Aquaculture,2010,309(1/2/3/4):245-252.DOI:10.1016/j.aquaculture.2010.08.022.
[12]JIA X H,EKMAN M,GROVE H,et al.Proteome changes in bovine longissimus thoracis muscle during the early postmortem storage period[J].Journal of Proteome Research,2007,6(7):2720-2731.DOI:10.1021/pr070173o.
[13]李强,刘永乐,王发祥,等.草鱼肌肉蛋白质在冷藏过程中的变化[J].食品科学,2013,34(3):55-58.
[14]LIU Yongle,LI Xianghong,ZHOU Xiaoling,et al.Effects of glutaminase deamidation on the structure and solubility of rice glutelin[J].LWT-Food Science and Technology,2011,44(10):2205-2210.DOI:10.1016/j.lwt.2011.05.011.
[15]曾茂茂,王霄,陈洁.蛋白质疏水性测定方法的相关性及适用性[J].食品科学,2011,32(15):117-120.
[16]姜杨,李婷婷,晋高伟,等.草鱼冷藏过程中新鲜度的综合评价[J].食品科学,2014,35(20):281-285.DOI:10.7506/spkx1002-6630-201420055.
[17]RIEBROY S,BENJAKUL S,VISESSANGUAN W,et al.Effect of iced storage of bigeye snapper(Priacanthus tayenus)on the chemical composition,properties and acceptability of Som-fug,a fermented Thai fish mince[J].Food Chemistry,2007,102(1):270-280.DOI:10.1016/j.foodchem.2006.05.017.
[18]李凯风,罗永康,冯启超,等.鱼鳞蛋白酶解物为基料的涂膜剂对鲫的保鲜效果[J].水产学报,2011,35(7):1113-1119.DOI:10.3724/SP.J.1231.2011.17339.
[19]杨胜平,谢晶,佟懿.不同浓度壳聚糖对带鱼冷藏保鲜效果的研究[J].山西农业科学,2010,38(2):77-80.DOI:I0.3969/j.issn.1002-2481.2010.02.22.
[20]老莹,胡文忠,冯可,等.天然抑菌剂的抑菌机理及其在果蔬保鲜中的应用[J].食品与发酵工业,2018,44(9):288-293.DOI:10.13995/j.cnki.11-1802/ts.016977.
[21]梁丽媚,李思东,李程鹏,等.壳聚糖及其衍生物抗菌活性的研究进展[J].广州化工,2017,45(20):6-9;15.
[22]FAN W J,CHI Y L,ZHANG S.The use of a tea polyphenol dip to extend the shelf life of silver carp(Hypophthalmicthys molitrix)during storage in ice[J].Food Chemistry,2008,108(1):148-153.
[23]CALLEWAERT L,MICHIELS C W.Lysozymes in the animal kingdom[J].Journal of Biosciences,2010,35(1):127-160.DOI:10.1007/s12038-010-0015-5.
[24]郑红,苏现波,马良,等.货架期冷藏过程中鳝鱼肉理化指标及蛋白质变化规律[J].食品科学,2018,39(7):215-220.DOI:10.7506/spkx1002-6630-201807032.
[25]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.DOI:10.1080/10408390591000929.
[26]YU Dawei,REGENSTEIN J M,ZANG Jinhong,et al.Inhibitory effects of chitosan-based coatings on endogenous enzyme activities,proteolytic degradation and texture softening of grass carp(Ctenopharyngodon idellus)fillets stored at 4℃[J].Food Chemistry,2018,262:1-6.DOI:10.1016/j.foodchem.2018.04.070.
[27]田悦,杜军保.二硫键和巯基在蛋白质结构功能中的作用及分析方法[J].实用儿科临床杂志,2007(19):1499-1501.
[28]BENJAKUL S,VISESSANGUAN W,THONGKAEW C,et al.Comparative study on physicochemical changes of muscle proteins from some tropical fish during frozen storage[J].Food Research International,2003,36(8):787-795.DOI:10.1016/S0963-9969(03)00073-5.
[29]LU Han,WANG Hang,LUO Yongkang.Changes in protein oxidation,water-holding capacity,and texture of bighead carp fillets under chilled and partial frozen storage[J].Journal of Aquatic Food Product Technology,2017,26(5):566-577.DOI:10.1080/10498850.2016.1238860.
[30]RAMIREZ J A,MARTIN-POLO M O,BANDMAN E.Fish myosin aggregation as affected by freezing and initial physical state[J].Journal of Food Science,2000,65(4):556-560.
[31]刘嘉莉,蓝蔚青,刘大勇,等.壳聚糖在水产品保鲜中应用研究进展[J].食品与机械,2019,35(3):231-236.DOI:10.13652/j.issn.1003-5788.2019.03.041.
[32]NAKAI S.Structure-function relationships of food proteins:with an emphasis on the importance of protein hydrophobicity[J].Journal of Agricultural and Food Chemistry,1983,31(4):676-683.
[33]吕卫金,赵进,汪金林,等.茶多酚延缓冷藏大黄鱼肌原纤维蛋白变性降解机理研究[J].中国食品学报,2014,14(1):60-67.DOI:10.16429/j.1009-7848.2014.01.017.
[34]李向红,王发祥,张付兰,等.冷藏过程中草鱼肌原纤维蛋白结构的变化[J].食品与发酵工业,2015,41(10):177-181.DOI:10.13995/j.cnki.11-1802/ts.201510033.
[35]BENJAKUL S,SEYMOUR T A,MORRISSEY M T,et al.Physicochemical changes in Pacific whiting muscle proteins during iced storage[J].Journal of Food Science,1997,62(4):729-733.