秘鲁鱿鱼肌原纤维蛋白质凝胶特性的研究
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摘要
秘鲁鱿鱼是美洲大赤鱿的俗称,学名为Dosidicus gigas,亦称jumboflying squid,属于真鱿科(Ommastrephidae)美洲大赤鱿属(Dosidicus)。该鱿鱼体形较大,成长迅速,是迄今为止发现的个体最大、资源最丰富的鱿鱼种类之一。但是秘鲁鱿鱼水分含量高,还含有不受人欢迎的酸、苦、涩味,大大影响了它的利用价值。目前对秘鲁鱿鱼的开发已成为国内水产加工的一个重要方向。鱼糜是经切碎、漂洗处理的鱼肉产品,是海洋食品加工中的一种重要功能性原料,其主要成分为盐溶性肌原纤维蛋白。本文以秘鲁鱿鱼和市售鱿鱼为研究对象,主要从鱿鱼肌原纤维蛋白的凝胶特性入手,系统地研究了其凝胶的特点,并从肌原纤维凝胶机理出发,研究改善肌原纤维蛋白凝胶特性的方法,进而为利用秘鲁鱿鱼加工优质冷冻鱼糜和鱼糜制品提供理论基础和技术指导。
研究结果表明:大秘鲁鱿鱼、中秘鲁鱿鱼和市售小鱿鱼的水分含量、灰分含量、粗蛋白含量差异较显著,但中鱿鱼与大鱿鱼的营养成分更接近。秘鲁鱿鱼肉中含有17种氨基酸,8种人体必需氨基酸俱全,秘鲁大鱿鱼的氨含量特别高,而秘鲁中鱿鱼和市售鱿鱼的氨含量则比较低。秘鲁鱿鱼中钾元素含量高,钙、铜元素含量比市售鱿鱼低,秘鲁鱿鱼中其他微量元素含量与市售鱿鱼含量差异较小。
大、中秘鲁鱿鱼的蛋白质组分较为相似,与市售小鱿鱼的蛋白质有较大差别。秘鲁鱿鱼肌原纤维Ca~(2+)-ATPase活性随保温时间、保温温度、离子强度的增加而显著降低(p＜0.05),肌原纤维的Ca~(2+)-ATPase稳定性变差,肌原纤维变性速度增快。添加Ca~(2+)能抑制秘鲁鱿鱼肌原纤维Ca~(2+)-ATPase的失活,提高肌原纤维的稳定性。
蛋白质、淀粉和阿拉伯胶等添加物对鱿鱼肌原纤维凝胶强度有显著影响。其中大豆蛋白和阿拉伯胶对秘鲁鱿鱼肌原纤维蛋白凝胶的影响更显著。正交试验分析得出,秘鲁大鱿鱼肌原纤维中添加大豆蛋白的最优化条件是添加4%大豆蛋白,pH值等于7,50℃加热1h;中鱿鱼肌原纤维中添加大豆蛋白的最优化条件是添加4%大豆蛋白,pH值为8,40℃加热1.5hrs;小鱿鱼肌原纤维中添加大豆蛋白的最优化条件是添加4%大豆蛋白,pH值为8,40℃加热1.5hrs。鱿鱼肌原纤维添加阿拉伯胶凝胶的最优化条件是pH值7,阿拉伯胶添加添加量是0.5%,凝胶温度50℃,凝胶时间1h。
酶添加量、TG作用时间、TG作用温度、TG作用pH对鱿鱼肌原纤维凝胶强度有显著影响。正交试验分析得出,鱿鱼肌原纤维凝胶化的最优化条件是添加2%的TG,pH值为7,40℃加热2hrs。
添加大豆蛋白和阿拉伯胶后,鱿鱼肌原纤维凝胶形成的三维网状结构更致密更均匀,多孔性和均一性更好。添加了TG后,鱿鱼肌原纤维凝胶所形成的三维网状结构更加致密,成孔性更好,孔隙更小更多,分布更加均匀,蛋白颗粒更细小。
超高压诱导肌原纤维蛋白凝胶化,通过凝胶强度测定和SDS-PAGE电泳以及ESEM及盒维数分析可以发现超高压凝胶比热诱导凝胶的硬度小,强度接近。加热处理比超高压处理肌动蛋白(AC)含量要高,而肌球蛋白含量低,热诱导凝胶促使肌球蛋白重链(MHC)形成交联键,对肌动蛋白没有什么影响,而超高压对肌动蛋白、肌动球蛋白和副肌球蛋白有解聚作用或对蛋白质有聚集凝集等变性的影响。
对超高压加工秘鲁鱿鱼鱼糜条件进行了优化,结果如下:
随着超高压处理压力的增大,秘鲁鱿鱼鱼糜凝胶强度呈先增后减的变化趋势,在300MPa～600MPa的压力下,超高压处理的凝胶强度都比热处理的要高,说明超高压比热处理在凝胶的形成上有更大的优势。
压力和保压时间对鱼糜凝胶硬度具有显著性影响(P＜0.05),两者的交互作用对硬度值也具有显著性影响(P＜0.05);压力对鱿鱼鱼糜的水分活度值无显著影响。先超高压凝胶化再热处理鱼糕化对鱿鱼鱼糜水分活度的影响不显著(P＞0.05)。
鱼糜凝胶强度随着马铃薯淀粉和大豆蛋白添加量的增大而增大,同一添加量时,先超高压凝胶化再热处理鱼糕化的效果均优于两段式热处理的效果。
超高压加工的鱼糜白度随马铃薯淀粉和大豆蛋白添加量的增大呈现先增大,后减小的趋势,而两段式热加工鱼糜的白度随着马铃薯淀粉添加量的增大而下降。
Peru Squid is the informal name of Dosidicus gigas,also named jumbo flying squid.It belongs to Dosidicus Ommastrephidae.It grew rapidly and had quite big shape.The highest concentration occurs off the Peruvian coast in the southern hemisphere and in the Gulf of California in the northern hemisphere.Over recent years,D.gigas became one of the biggest and most abundant cephalopod fisheries in the Pacific Ocean, mainly in Peru and Mexico.But high moisture and disagreeable taste in the muscle of D.gigas utilization of this cephalopod resource.Surimi and surimi-based product are good choice because the disagreeable taste could be removed during the process.The bases of surimi and surimi-based product are myofibrils dissolved in salt solution.The objects of this research were Peru Squid and normal squid sold in market.Myofibrils of squid were extracted and the gel properties were studied to improve gelation capacity of myofibrils.From the concultion metioned above,Peru squid can be a new material for high quality surimi.
The result showed giant Peru squid(Carcass weight＞6kg),medium Peru squid(Carcass weight between 3～6kg)and little squid(normal squid sold in market,carcass weight＜1kg)had significance of difference in water content,ashes content and crude protein content.Ammonia content in giant Peru squid was much higher than the other which made giant Peru squid a strange astringent and acerbic flavour.There were more kalium content and less copper,calcium content in Peru squid than little squid and there were no significant difference in other trace element content among the three size squid.
Similar protein constitute were observed in giant and medium Peru squid and different with little squid.Time,incubation temperature and ionic intensity showed significant effect on the activity of Ca~(2+)-ATPase in Peru squid myofibrils(p＜0.05).Stability of Ca-ATPase could be improved by certain content of sodium chloride.
Additives such as nonmuscle protein,starch and Acacia Gum showed significant influence to squid myofibrils gel strength in which soy protein and Acacia Gum exhibited salient effect.Operational conditions assay achieved optimization of giant squid myofibrils gellation adding nonmuscle protein were 4%w/w soy protein,pH 7,50℃heated for 1hrs and 4%w/w soy protein,pH 8,40℃heated for 1.5hrs for both medium and little squid. Operational conditions assay achieved optimization of squid myofibrils gellation adding Acacia Gum protein were 0.5%w/w concentration,pH 7, 50℃heated for 1hrs.
TG concentration,time,incubation temperature and pH showed significant influence to squid myofibrils gel strength.Operational conditions assay achieved optimization of squid myofibrils gellation with TG were 2%w/w TG,pH 7,40℃heated for 2hrs.
The microstructure showed that the clusters network of squid myofibrils added with soy protein,Acacia Gum and TG were composed by smaller aggregates than those control.The microstructure showed that both the size of the granules and the cavities among clusters seemed to be larger with the TG concentration decreased.
Effect of high pressure on gel properties was studied.The result showed gels induced by 300MPa and 400MPa demonstrated better gel strength than other pressure treated gels.Microstructure of surimi gels induced by different high pressure showed significance of difference and exhibited a compact and homogeneous network at 300MPa and 400MPa.Gel hardness of high pressure-induced gels were less than heat-induced(40℃,2h and 90℃,30 min)gels and springness of high pressure-induced gels were higher than heat-induced(40℃,2h and 90℃, 30 min)gels and so gel formed by this two method showed similar gel strength.SDS-PAGE showed high pressure prompted more Actin and Paramyosin aggregating and cross-linked with Myosin or filled into the net-work formed by Myosin heavy chain than heat treated.
Operational conditions for Peru squid surimi processed by high pressure were studied.The result showed:
Surimi gel strength made from Peru squid was larger as the pressure increasing.Maximum of gel strength arrived at 500MPa,15min which was 7 times to heat-induced(40℃,2h)gels.Gel strength of surimi by high pressure between 300MPa～600MPa all higher than heat treated gels.
Surimi gel strength of surimi gels processed by high pressure(300 MPa～600 MPa,10～20min)followed by heat treated(90℃,30 min)were higher than heat treated gels(40℃,2h and 90℃,30 min).
Activity wet of giant squid surimi exhibited no significant difference as the pressure changed.
The more potato starch added into surimi the higher gel strength.Gel strength of high pressure-induced gels were obviously higher than heat-induced gels with 4%or 6%w/w potato starch.
Gel strength of surimi gels adding soy protein increaced along with the rising of pressure.Gel properties of surimi obtained from adding 6%or 8%w/w soy protein and treated by more than 350 MPa were satisfactory.
Whiteness of surimi treated by high pressure followed heat showed no significant difference between gels with 6%and 8%w/w potato starch (P＞0.05)while the whiteness of heat-induced gel dropped as the concentration of potato starch rised.
Whiteness of surimi achieved maximum treated at 400MPa.Surimi properties were satisfied in these processing conditions.
研究结果表明:大秘鲁鱿鱼、中秘鲁鱿鱼和市售小鱿鱼的水分含量、灰分含量、粗蛋白含量差异较显著,但中鱿鱼与大鱿鱼的营养成分更接近。秘鲁鱿鱼肉中含有17种氨基酸,8种人体必需氨基酸俱全,秘鲁大鱿鱼的氨含量特别高,而秘鲁中鱿鱼和市售鱿鱼的氨含量则比较低。秘鲁鱿鱼中钾元素含量高,钙、铜元素含量比市售鱿鱼低,秘鲁鱿鱼中其他微量元素含量与市售鱿鱼含量差异较小。
大、中秘鲁鱿鱼的蛋白质组分较为相似,与市售小鱿鱼的蛋白质有较大差别。秘鲁鱿鱼肌原纤维Ca~(2+)-ATPase活性随保温时间、保温温度、离子强度的增加而显著降低(p＜0.05),肌原纤维的Ca~(2+)-ATPase稳定性变差,肌原纤维变性速度增快。添加Ca~(2+)能抑制秘鲁鱿鱼肌原纤维Ca~(2+)-ATPase的失活,提高肌原纤维的稳定性。
蛋白质、淀粉和阿拉伯胶等添加物对鱿鱼肌原纤维凝胶强度有显著影响。其中大豆蛋白和阿拉伯胶对秘鲁鱿鱼肌原纤维蛋白凝胶的影响更显著。正交试验分析得出,秘鲁大鱿鱼肌原纤维中添加大豆蛋白的最优化条件是添加4%大豆蛋白,pH值等于7,50℃加热1h;中鱿鱼肌原纤维中添加大豆蛋白的最优化条件是添加4%大豆蛋白,pH值为8,40℃加热1.5hrs;小鱿鱼肌原纤维中添加大豆蛋白的最优化条件是添加4%大豆蛋白,pH值为8,40℃加热1.5hrs。鱿鱼肌原纤维添加阿拉伯胶凝胶的最优化条件是pH值7,阿拉伯胶添加添加量是0.5%,凝胶温度50℃,凝胶时间1h。
酶添加量、TG作用时间、TG作用温度、TG作用pH对鱿鱼肌原纤维凝胶强度有显著影响。正交试验分析得出,鱿鱼肌原纤维凝胶化的最优化条件是添加2%的TG,pH值为7,40℃加热2hrs。
添加大豆蛋白和阿拉伯胶后,鱿鱼肌原纤维凝胶形成的三维网状结构更致密更均匀,多孔性和均一性更好。添加了TG后,鱿鱼肌原纤维凝胶所形成的三维网状结构更加致密,成孔性更好,孔隙更小更多,分布更加均匀,蛋白颗粒更细小。
超高压诱导肌原纤维蛋白凝胶化,通过凝胶强度测定和SDS-PAGE电泳以及ESEM及盒维数分析可以发现超高压凝胶比热诱导凝胶的硬度小,强度接近。加热处理比超高压处理肌动蛋白(AC)含量要高,而肌球蛋白含量低,热诱导凝胶促使肌球蛋白重链(MHC)形成交联键,对肌动蛋白没有什么影响,而超高压对肌动蛋白、肌动球蛋白和副肌球蛋白有解聚作用或对蛋白质有聚集凝集等变性的影响。
对超高压加工秘鲁鱿鱼鱼糜条件进行了优化,结果如下:
随着超高压处理压力的增大,秘鲁鱿鱼鱼糜凝胶强度呈先增后减的变化趋势,在300MPa～600MPa的压力下,超高压处理的凝胶强度都比热处理的要高,说明超高压比热处理在凝胶的形成上有更大的优势。
压力和保压时间对鱼糜凝胶硬度具有显著性影响(P＜0.05),两者的交互作用对硬度值也具有显著性影响(P＜0.05);压力对鱿鱼鱼糜的水分活度值无显著影响。先超高压凝胶化再热处理鱼糕化对鱿鱼鱼糜水分活度的影响不显著(P＞0.05)。
鱼糜凝胶强度随着马铃薯淀粉和大豆蛋白添加量的增大而增大,同一添加量时,先超高压凝胶化再热处理鱼糕化的效果均优于两段式热处理的效果。
超高压加工的鱼糜白度随马铃薯淀粉和大豆蛋白添加量的增大呈现先增大,后减小的趋势,而两段式热加工鱼糜的白度随着马铃薯淀粉添加量的增大而下降。
Peru Squid is the informal name of Dosidicus gigas,also named jumbo flying squid.It belongs to Dosidicus Ommastrephidae.It grew rapidly and had quite big shape.The highest concentration occurs off the Peruvian coast in the southern hemisphere and in the Gulf of California in the northern hemisphere.Over recent years,D.gigas became one of the biggest and most abundant cephalopod fisheries in the Pacific Ocean, mainly in Peru and Mexico.But high moisture and disagreeable taste in the muscle of D.gigas utilization of this cephalopod resource.Surimi and surimi-based product are good choice because the disagreeable taste could be removed during the process.The bases of surimi and surimi-based product are myofibrils dissolved in salt solution.The objects of this research were Peru Squid and normal squid sold in market.Myofibrils of squid were extracted and the gel properties were studied to improve gelation capacity of myofibrils.From the concultion metioned above,Peru squid can be a new material for high quality surimi.
The result showed giant Peru squid(Carcass weight＞6kg),medium Peru squid(Carcass weight between 3～6kg)and little squid(normal squid sold in market,carcass weight＜1kg)had significance of difference in water content,ashes content and crude protein content.Ammonia content in giant Peru squid was much higher than the other which made giant Peru squid a strange astringent and acerbic flavour.There were more kalium content and less copper,calcium content in Peru squid than little squid and there were no significant difference in other trace element content among the three size squid.
Similar protein constitute were observed in giant and medium Peru squid and different with little squid.Time,incubation temperature and ionic intensity showed significant effect on the activity of Ca~(2+)-ATPase in Peru squid myofibrils(p＜0.05).Stability of Ca-ATPase could be improved by certain content of sodium chloride.
Additives such as nonmuscle protein,starch and Acacia Gum showed significant influence to squid myofibrils gel strength in which soy protein and Acacia Gum exhibited salient effect.Operational conditions assay achieved optimization of giant squid myofibrils gellation adding nonmuscle protein were 4%w/w soy protein,pH 7,50℃heated for 1hrs and 4%w/w soy protein,pH 8,40℃heated for 1.5hrs for both medium and little squid. Operational conditions assay achieved optimization of squid myofibrils gellation adding Acacia Gum protein were 0.5%w/w concentration,pH 7, 50℃heated for 1hrs.
TG concentration,time,incubation temperature and pH showed significant influence to squid myofibrils gel strength.Operational conditions assay achieved optimization of squid myofibrils gellation with TG were 2%w/w TG,pH 7,40℃heated for 2hrs.
The microstructure showed that the clusters network of squid myofibrils added with soy protein,Acacia Gum and TG were composed by smaller aggregates than those control.The microstructure showed that both the size of the granules and the cavities among clusters seemed to be larger with the TG concentration decreased.
Effect of high pressure on gel properties was studied.The result showed gels induced by 300MPa and 400MPa demonstrated better gel strength than other pressure treated gels.Microstructure of surimi gels induced by different high pressure showed significance of difference and exhibited a compact and homogeneous network at 300MPa and 400MPa.Gel hardness of high pressure-induced gels were less than heat-induced(40℃,2h and 90℃,30 min)gels and springness of high pressure-induced gels were higher than heat-induced(40℃,2h and 90℃, 30 min)gels and so gel formed by this two method showed similar gel strength.SDS-PAGE showed high pressure prompted more Actin and Paramyosin aggregating and cross-linked with Myosin or filled into the net-work formed by Myosin heavy chain than heat treated.
Operational conditions for Peru squid surimi processed by high pressure were studied.The result showed:
Surimi gel strength made from Peru squid was larger as the pressure increasing.Maximum of gel strength arrived at 500MPa,15min which was 7 times to heat-induced(40℃,2h)gels.Gel strength of surimi by high pressure between 300MPa～600MPa all higher than heat treated gels.
Surimi gel strength of surimi gels processed by high pressure(300 MPa～600 MPa,10～20min)followed by heat treated(90℃,30 min)were higher than heat treated gels(40℃,2h and 90℃,30 min).
Activity wet of giant squid surimi exhibited no significant difference as the pressure changed.
The more potato starch added into surimi the higher gel strength.Gel strength of high pressure-induced gels were obviously higher than heat-induced gels with 4%or 6%w/w potato starch.
Gel strength of surimi gels adding soy protein increaced along with the rising of pressure.Gel properties of surimi obtained from adding 6%or 8%w/w soy protein and treated by more than 350 MPa were satisfactory.
Whiteness of surimi treated by high pressure followed heat showed no significant difference between gels with 6%and 8%w/w potato starch (P＞0.05)while the whiteness of heat-induced gel dropped as the concentration of potato starch rised.
Whiteness of surimi achieved maximum treated at 400MPa.Surimi properties were satisfied in these processing conditions.
引文
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