干露对活品虾夷扇贝闭壳肌SOD酶活性及其性质的影响
|
摘要
为探究活品贝类在流通过程中的品质变化与胁迫代谢的关系,以虾夷扇贝Patinopecten yessoensis为研究对象,将采捕后的活品扇贝分为3组,进行冷却及室温干藏处理,其中冷却干藏温度为4℃,分充氧包装组(CDo)和充气包装组(CDa),常温环境温度20℃为充气包装组(ADa),对3个处理组进行跟踪观察,并每日采样直至扇贝死亡。探究其闭壳肌粗酶提取液中超氧化物歧化酶(SOD)的变化规律及其酶学性质。结果表明:缺氧和高温导致扇贝加速死亡,冷却充氧包装组(CDo)在第6天全部死亡,冷却充气包装组(CDa)在第4天全部死亡,而常温充气包装组(ADa)则在第2天就全部死亡;冷却干露贮藏期间,闭壳肌水溶性蛋白质含量出现轻微下降,SDS-PAGE电泳分析显示,卵清蛋白(相对分子质量44000)条带附近的蛋白组分出现降解;扇贝闭壳肌中SOD活性与胁迫出现应答关联,常温组(ADa)SOD活性高于冷却组(CDo、CDa),充气组CDa又高于充氧组CDo,干露期间所有处理组的SOD活性整体呈现上升趋势;扇贝闭壳肌中SOD同样具有较高的耐热性,耐热温度为60℃,其p H稳定范围为中性,Cu~(2+)、Zn~(2+)、Fe~(3+)、十二烷基硫酸钠(SDS)、柠檬酸和H_2O_2等对SOD活性有抑制作用,低浓度Mn~(2+)对SOD活性有促进作用,高浓度Mn~(2+)则有抑制作用。初步研究表明,干露与活品虾夷扇贝中SOD活性有一定相关性,温度、缺氧等胁迫均引起虾夷扇贝闭壳肌中SOD活性应答,可以进行深入研究。
Activity and characteristics of superoxide dismutase( SOD) in an adductor were determined in yesso scallop Patinopecten yessoensis exposed to chilled dry storage with oxygen packaged( CDo) and air packaged( CDa)at 4 ℃,and ambient dry storage with air packaged( ADa) at 20 ℃ to explore the quality mechanism of post-harvest live scallop and metabolic suppression during live dry storage. High mortality due to hypoxia and higher temperature were observed in the scallop in group ADa,while prolonged survival period was found in the scallop exposed to chilling treatment in group CDo. During chilling exposure,the water-soluble protein content of the adductor was decreased slightly,and SDS-PAGE showed that the protein nearby the ovalbumin( 44 000) was to some extent degraded. There was higher activity of SOD in the scallop adductor in group ADa than that in groups CDo and CDa,higher in group CDa than that in group CDo,though the SOD activity was shown to be increased in all groups during the dry storage. SOD activity was found to be heat stable in adductor of the scallop at heat-stable temperature of60 ℃ and inhibited by Cu~(2+),Zn~(2+),Fe~(3+),SDS,citric acid and H_2O_2. However,the SOD activity was promoted at low concentration of Mn~(2+)and inhibited at high concentration of Mn~(2+). The findings indicate that there is correlation between the anaerobic stress and the superoxide dismutase activity in live scallop adductor muscle,and that further investigation is required to clarify this mechanism.
Activity and characteristics of superoxide dismutase( SOD) in an adductor were determined in yesso scallop Patinopecten yessoensis exposed to chilled dry storage with oxygen packaged( CDo) and air packaged( CDa)at 4 ℃,and ambient dry storage with air packaged( ADa) at 20 ℃ to explore the quality mechanism of post-harvest live scallop and metabolic suppression during live dry storage. High mortality due to hypoxia and higher temperature were observed in the scallop in group ADa,while prolonged survival period was found in the scallop exposed to chilling treatment in group CDo. During chilling exposure,the water-soluble protein content of the adductor was decreased slightly,and SDS-PAGE showed that the protein nearby the ovalbumin( 44 000) was to some extent degraded. There was higher activity of SOD in the scallop adductor in group ADa than that in groups CDo and CDa,higher in group CDa than that in group CDo,though the SOD activity was shown to be increased in all groups during the dry storage. SOD activity was found to be heat stable in adductor of the scallop at heat-stable temperature of60 ℃ and inhibited by Cu~(2+),Zn~(2+),Fe~(3+),SDS,citric acid and H_2O_2. However,the SOD activity was promoted at low concentration of Mn~(2+)and inhibited at high concentration of Mn~(2+). The findings indicate that there is correlation between the anaerobic stress and the superoxide dismutase activity in live scallop adductor muscle,and that further investigation is required to clarify this mechanism.
引文
[1]农业部渔业渔政管理局.中国渔业统计年鉴2017[M].北京:中国农业出版社,2017.
[2]刘金洋,刘俊荣,田元勇,等.捕后处置对活品底播虾夷扇贝生化代谢的影响[J].水产学报,2017,41(1):81-87.
[3]刘金洋.虾夷扇贝在活品流通过程中的代谢变化[D].大连:大连海洋大学,2016.
[4]杨婷婷,刘俊荣,俞微微,等.活品流通过程中虾夷扇贝风味品质的变化[J].水产学报,2015,39(1):136-146.
[5]傅润泽,沈建,王锡昌.底播虾夷扇贝活品流通前后挥发性成分的对比分析[J].食品科学,2015,36(2):110-113.
[6]张克烽,张子平,陈芸,等.动物抗氧化系统中主要抗氧化酶基因的研究进展[J].动物学杂志,2007,42(2):153-160.
[7]林庆斌,廖升荣,熊亚红,等.超氧化物歧化酶(SOD)的研究和应用进展[J].化学世界,2006,47(6):378-381.
[8]周作强,丁晴晴,刘其根,等.缺氧对贝类的胁迫效应及对其免疫系统的影响[J].生态毒理学报,2013,8(3):324-330.
[9]孙虎山,李光友.双壳贝类参与免疫防御的体液因子[J].海洋科学,2001,25(4):34-36.
[10]郑尧,刘俊荣,周晏琳,等.捕后虾夷扇贝闭壳肌免疫因子与活品品质评价初探[J].大连海洋大学学报,2017,32(2):217-223.
[11]路苹,于同泉,王淑英,等.蛋白质测定方法评价[J].北京农学院学报,2006,21(2):65-69.
[12]张龙,刘俊荣,田元勇,等.菲律宾蛤仔(Ruditapes philippinarum)软体部位含氮物的组成及其分布[J].食品科学,2015,36(24):95-100.
[13]桂兴芬,阎家麒,朱建梅.几种改进的邻苯三酚自氧化法测定超氧化物歧化酶活性的比较[J].中国兽医科学,1993,23(1):23-24.
[14]高若梅,邹洪,袁倬斌.用电化学方法研究邻苯三酚的自氧化机理[J].分析化学,1997,25(3):297-300.
[15] Rupp G S,Parsons G J.Effects of salinity and temperature on the survival and byssal attachment of the lion's paw scallop Nodipecten nodosus at its southern distribution limit[J].Journal of Experimental Marine Biology and Ecology,2004,309(2):173-198.
[16] Lee A C,Lee K T.The enzyme activities of opine and lactate dehydrogenases in the gills,mantle,foot,and adductor of the hard clam Meretrix lusoria[J].Journal of Marine Science and Technology,2011,19(4):361-367.
[17] De Almeida E A,Celso Dias Bainy A,De Melo Loureiro A P,et al.Oxidative stress in Perna perna and other bivalves as indicators of environmental stress in the Brazilian marine environment:antioxidants,lipid peroxidation and DNA damage[J]. Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,2007,146(4):588-600.
[18] Yamashita M,Konagaya S.High activities of cathepsins B,D,H,and L in the white muscle of chum salmon in spawning migration[J].Comparative Biochemistry and Physiology Part B:Comparative Biochemistry,1990,95(1):149-152.
[19]王跃军,孙谧,李民,等.扇贝超氧化物歧化酶的纯化及性质研究[J].海洋水产研究,1998,19(2):69-75.
[20]吴燕燕,李来好,郝志明,等.罗非鱼肝脏中超氧化物歧化酶的提取、纯化与分析[J].水产学报,2007,31(4):518-524.
[21]王晓宇.四角蛤蜊Mactra veneriformis对镉和汞污染胁迫的生理响应[D].北京:中国科学院研究生院(海洋研究所),2009.
[22]赵艳芳,吴继法,翟毓秀,等.镉胁迫对不同镉富集能力海水养殖贝类抗氧化能力的影响——以扇贝和菲律宾蛤仔为例[J].生态毒理学报,2014,9(2):224-232.
[23]毕研娇.白泥脱硫废水排海的生态风险评估[D].青岛:中国海洋大学,2012.
[24]王清.几种重金属和有机污染物对文蛤Meretrxi meretrix生态毒理效应的研究[D].北京:中国科学院研究生院(海洋研究所),2010.
[25]吕昊泽.缢蛏、光滑河蓝蛤和河蚬对盐度的适应性及碳、氮收支研究[D].上海:上海海洋大学,2014.
[26]肖湘,俞丽君,郑扬明,等.贻贝超氧化物歧化酶的分离纯化和部分性质[J].台湾海峡,1999,18(2):177-180.
[27]沈洪国,唐云明,韩晨霞.黄鳝内脏铁型超氧化物歧化酶的纯化[J].水产科学,2005,24(2):15-17.
[28]王伟伟,刘存歧,李道季,等.3种虾超氧化物歧化酶部分理化性质比较[J].水产科学,2009,28(4):200-204.
[29]孙虎山,李光友.栉孔扇贝血淋巴中超氧化物歧化酶和过氧化氢酶活性及其性质的研究[J].海洋与湖沼,2000,31(3):259-265.
[30]高梦祥,杨凤琴.冬瓜多酚氧化酶的特性及抑制剂研究[J].食品科技,2008,33(2):11-14.
[31]王凡,赵元凤,吕景才,等.铜污染对扇贝内脏团抗氧化酶活性的影响[J].水产科学,2008,27(12):622-624.
[32]石芳芳.栉孔扇贝和海湾扇贝免疫防御特性的比较研究[D].北京:中国科学院研究生院(海洋研究所),2006.
[33] Pan Luqing,Ren Jiayun,Zheng Debin.Effects of benzo(a)pyrene exposure on the antioxidant enzyme activity of scallop Chlamys farreri[J]. Chinese Journal of Oceanology and Limnology,2009,27(1):43-53.
[34] Wang Xingqiang,Wang Lingling,Zhang Hua,et al. Immune response and energy metabolism of Chlamys farreri under Vibrio anguillarum challenge and high temperature exposure[J]. Fish&Shellfish Immunology,2012,33(4):1016-1026.
[35]高振锟,张继红,李敏,等.温度波动对不同规格虾夷扇贝(Patinopecten yessoensis)生理和免疫指标的影响[J].渔业科学进展,2017,38(3):148-154.
[2]刘金洋,刘俊荣,田元勇,等.捕后处置对活品底播虾夷扇贝生化代谢的影响[J].水产学报,2017,41(1):81-87.
[3]刘金洋.虾夷扇贝在活品流通过程中的代谢变化[D].大连:大连海洋大学,2016.
[4]杨婷婷,刘俊荣,俞微微,等.活品流通过程中虾夷扇贝风味品质的变化[J].水产学报,2015,39(1):136-146.
[5]傅润泽,沈建,王锡昌.底播虾夷扇贝活品流通前后挥发性成分的对比分析[J].食品科学,2015,36(2):110-113.
[6]张克烽,张子平,陈芸,等.动物抗氧化系统中主要抗氧化酶基因的研究进展[J].动物学杂志,2007,42(2):153-160.
[7]林庆斌,廖升荣,熊亚红,等.超氧化物歧化酶(SOD)的研究和应用进展[J].化学世界,2006,47(6):378-381.
[8]周作强,丁晴晴,刘其根,等.缺氧对贝类的胁迫效应及对其免疫系统的影响[J].生态毒理学报,2013,8(3):324-330.
[9]孙虎山,李光友.双壳贝类参与免疫防御的体液因子[J].海洋科学,2001,25(4):34-36.
[10]郑尧,刘俊荣,周晏琳,等.捕后虾夷扇贝闭壳肌免疫因子与活品品质评价初探[J].大连海洋大学学报,2017,32(2):217-223.
[11]路苹,于同泉,王淑英,等.蛋白质测定方法评价[J].北京农学院学报,2006,21(2):65-69.
[12]张龙,刘俊荣,田元勇,等.菲律宾蛤仔(Ruditapes philippinarum)软体部位含氮物的组成及其分布[J].食品科学,2015,36(24):95-100.
[13]桂兴芬,阎家麒,朱建梅.几种改进的邻苯三酚自氧化法测定超氧化物歧化酶活性的比较[J].中国兽医科学,1993,23(1):23-24.
[14]高若梅,邹洪,袁倬斌.用电化学方法研究邻苯三酚的自氧化机理[J].分析化学,1997,25(3):297-300.
[15] Rupp G S,Parsons G J.Effects of salinity and temperature on the survival and byssal attachment of the lion's paw scallop Nodipecten nodosus at its southern distribution limit[J].Journal of Experimental Marine Biology and Ecology,2004,309(2):173-198.
[16] Lee A C,Lee K T.The enzyme activities of opine and lactate dehydrogenases in the gills,mantle,foot,and adductor of the hard clam Meretrix lusoria[J].Journal of Marine Science and Technology,2011,19(4):361-367.
[17] De Almeida E A,Celso Dias Bainy A,De Melo Loureiro A P,et al.Oxidative stress in Perna perna and other bivalves as indicators of environmental stress in the Brazilian marine environment:antioxidants,lipid peroxidation and DNA damage[J]. Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,2007,146(4):588-600.
[18] Yamashita M,Konagaya S.High activities of cathepsins B,D,H,and L in the white muscle of chum salmon in spawning migration[J].Comparative Biochemistry and Physiology Part B:Comparative Biochemistry,1990,95(1):149-152.
[19]王跃军,孙谧,李民,等.扇贝超氧化物歧化酶的纯化及性质研究[J].海洋水产研究,1998,19(2):69-75.
[20]吴燕燕,李来好,郝志明,等.罗非鱼肝脏中超氧化物歧化酶的提取、纯化与分析[J].水产学报,2007,31(4):518-524.
[21]王晓宇.四角蛤蜊Mactra veneriformis对镉和汞污染胁迫的生理响应[D].北京:中国科学院研究生院(海洋研究所),2009.
[22]赵艳芳,吴继法,翟毓秀,等.镉胁迫对不同镉富集能力海水养殖贝类抗氧化能力的影响——以扇贝和菲律宾蛤仔为例[J].生态毒理学报,2014,9(2):224-232.
[23]毕研娇.白泥脱硫废水排海的生态风险评估[D].青岛:中国海洋大学,2012.
[24]王清.几种重金属和有机污染物对文蛤Meretrxi meretrix生态毒理效应的研究[D].北京:中国科学院研究生院(海洋研究所),2010.
[25]吕昊泽.缢蛏、光滑河蓝蛤和河蚬对盐度的适应性及碳、氮收支研究[D].上海:上海海洋大学,2014.
[26]肖湘,俞丽君,郑扬明,等.贻贝超氧化物歧化酶的分离纯化和部分性质[J].台湾海峡,1999,18(2):177-180.
[27]沈洪国,唐云明,韩晨霞.黄鳝内脏铁型超氧化物歧化酶的纯化[J].水产科学,2005,24(2):15-17.
[28]王伟伟,刘存歧,李道季,等.3种虾超氧化物歧化酶部分理化性质比较[J].水产科学,2009,28(4):200-204.
[29]孙虎山,李光友.栉孔扇贝血淋巴中超氧化物歧化酶和过氧化氢酶活性及其性质的研究[J].海洋与湖沼,2000,31(3):259-265.
[30]高梦祥,杨凤琴.冬瓜多酚氧化酶的特性及抑制剂研究[J].食品科技,2008,33(2):11-14.
[31]王凡,赵元凤,吕景才,等.铜污染对扇贝内脏团抗氧化酶活性的影响[J].水产科学,2008,27(12):622-624.
[32]石芳芳.栉孔扇贝和海湾扇贝免疫防御特性的比较研究[D].北京:中国科学院研究生院(海洋研究所),2006.
[33] Pan Luqing,Ren Jiayun,Zheng Debin.Effects of benzo(a)pyrene exposure on the antioxidant enzyme activity of scallop Chlamys farreri[J]. Chinese Journal of Oceanology and Limnology,2009,27(1):43-53.
[34] Wang Xingqiang,Wang Lingling,Zhang Hua,et al. Immune response and energy metabolism of Chlamys farreri under Vibrio anguillarum challenge and high temperature exposure[J]. Fish&Shellfish Immunology,2012,33(4):1016-1026.
[35]高振锟,张继红,李敏,等.温度波动对不同规格虾夷扇贝(Patinopecten yessoensis)生理和免疫指标的影响[J].渔业科学进展,2017,38(3):148-154.