采用复合酶控制水解青鳞鱼蛋白的研究
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摘要
本文以青鳞鱼蛋白为原料，用复合酶控制水解技术制备蛋白水解物并对其进行了食品功能特性及抗骨质疏松活性研究。
在原料特性的研究中，主要研究了原料蛋白质和分离组分氨基酸构成。结果表明青鳞鱼蛋白含18种氨基酸(呈味氨基酸占44.0％、疏水氨基酸占41.9％)，其组分分子大小为肌浆蛋白＞基质蛋白＞碱溶性蛋白＞肌原纤维蛋白。由于基质蛋白高达52.46％、肌原纤维蛋白仅为4.19％，故青鳞鱼不适合制作鱼糜制品，但适合制作功能食品基料、明胶、活性肽制品及海鲜调味料等。
在木瓜蛋白酶、风味酶等复合酶控制水解过程中，首先采用正交试验确立了水解时间为120min时酶对青鳞鱼蛋白质的最适水解条件。其次，研究了以水解时间为变量因子的三种酶的水解产物。结果表明，复合酶用量比其2种前体酶减少了22倍，最适宜pH接近鱼肉水悬液自然pH；它在最适宜条件下存在三组线性关系，即：DH=0.0261t+8.8486(t=10-210min，r=0.9834)、MLPL=-0.02411t+11.5616(t≠0，r=-0.9369)和TFAA=0.1558t+38.76(t=30~210min，r=0.9891)；它对胱氨酸和苯丙氨酸+酪氨酸表现出高底物特异性，而对2种前体酶都不能水解的色氨酸连接的肽键具有一定的水解能力。
按照研究和实验分析，得出复合酶控制水解动力学模型及相关参数，分别为：DH=5.8821n[1+(0.05362e_0／s_0-0.0032)t]和R=(0.3154e_0-0.0186s_0)exp(-0.170DH)，复合酶失活常数K_d=0.0536min~(-1)，复合酶水解反应能够顺利进行的条件：e_0／s_0＞c_0(其中常数c_0=5.90×10~(-2))。
采用复合酶控制水解可大大改善青鳞鱼蛋白的溶解性，水解产物的NSI为89.3％-98.5％而且在酸性pH范围NSI略高于碱性pH范围；复合酶可以显著提高蛋白质的乳化活力，水解产物在酸性范围具有相对恒定且最高乳化活力指标，在碱性范围则表现出较低的乳化活力；随着DH的增加，最高乳化活力指标表现的pH范围更宽广，但都保持在112m~2／g以上。复合酶对乳化稳定性有降低趋势，在酸性范围比碱性范围具有相对较高的乳化稳定性；同时，较低DH的水解产物比较高DH的具有相对较低的抗热处理能力。蛋白质浓度为1％~5％时，FP随浓度的增加而增加；泡沫形成后的1min内，1％蛋白质浓度的泡沫较3％和5％的稳定，而15min后，泡沫的稳定性随蛋白质浓度的增加而增加。浓度为3％、DH为19.9％的水解产物，pH4.0时具有最低的起泡性能，而pH10.0时具有最高的起泡能力；pH4.0和8.0是泡沫最不稳定的pH点，而pH10.0时泡沫最稳定。
In this paper, studies on preparation of hydrolysate by use of controlled complicated-enzymes hydrolysis of Harengula Zunasi Bleeker proteins, food functionality and anti- osteoporosis action were carried out.
The studies on material protein peculiarity and its protein fraction showed that the protein of Harengula Zunasi Bleeker can be used in manufacturing functional food, gelatin, bioactive peptides and sea fresh condiment except for fish filling because the ratios of the content of taste amino acids and the content of hydrophobic amino acids to the total content of amino acids in protein were 44.0% and 41.9%, respectively, and that the ratios of the content of stroma protein and the content of myofibrilar protein to the total protein were 52.46% and 4. 19%, respectively. Meanwhile the molecule size of protein fraction was myogen> stroma protein> alkali-soluble protein > myofibrilar protein.
In order to establish the process of the controlled-complicated enzymes hydrolysis, first the optimum conditions of papain, flavorease and complicated enzymes were established by orthogonal trial with the constant time of 2 hours, secondly, the obtained hydrolysates by them at varying hydrolysis time were studied. The results showed that the usage of the complicated enzymes obtained by papain and flavorease may be decreased 22 times than its pre-enzyme and its optimum pH approaches to the pH of fish meat dissolved in water; that the hydrolysis course for the complicated enzymes is described with 3 linear relationship being expressed as follows: DH=0. 0261t+8. 8486(t = 10-210min, r=0. 9834) , MLPL=-0. 0241H + 11. 5616 ( t≠0, r=-0. 9369) and TFAA=0. 1558t+38. 76 (t = 30-210min, r=0. 9891) ; and the complicated enzymes has the strongest hydrolysis specificity toward the peptide linkages supplied by cysteine, phenylalanine and tyrosine and it can break the bond supplied by tryptophane, but papain and flavorease can not.
The kinetic model and relative parameter of
在原料特性的研究中，主要研究了原料蛋白质和分离组分氨基酸构成。结果表明青鳞鱼蛋白含18种氨基酸(呈味氨基酸占44.0％、疏水氨基酸占41.9％)，其组分分子大小为肌浆蛋白＞基质蛋白＞碱溶性蛋白＞肌原纤维蛋白。由于基质蛋白高达52.46％、肌原纤维蛋白仅为4.19％，故青鳞鱼不适合制作鱼糜制品，但适合制作功能食品基料、明胶、活性肽制品及海鲜调味料等。
在木瓜蛋白酶、风味酶等复合酶控制水解过程中，首先采用正交试验确立了水解时间为120min时酶对青鳞鱼蛋白质的最适水解条件。其次，研究了以水解时间为变量因子的三种酶的水解产物。结果表明，复合酶用量比其2种前体酶减少了22倍，最适宜pH接近鱼肉水悬液自然pH；它在最适宜条件下存在三组线性关系，即：DH=0.0261t+8.8486(t=10-210min，r=0.9834)、MLPL=-0.02411t+11.5616(t≠0，r=-0.9369)和TFAA=0.1558t+38.76(t=30~210min，r=0.9891)；它对胱氨酸和苯丙氨酸+酪氨酸表现出高底物特异性，而对2种前体酶都不能水解的色氨酸连接的肽键具有一定的水解能力。
按照研究和实验分析，得出复合酶控制水解动力学模型及相关参数，分别为：DH=5.8821n[1+(0.05362e_0／s_0-0.0032)t]和R=(0.3154e_0-0.0186s_0)exp(-0.170DH)，复合酶失活常数K_d=0.0536min~(-1)，复合酶水解反应能够顺利进行的条件：e_0／s_0＞c_0(其中常数c_0=5.90×10~(-2))。
采用复合酶控制水解可大大改善青鳞鱼蛋白的溶解性，水解产物的NSI为89.3％-98.5％而且在酸性pH范围NSI略高于碱性pH范围；复合酶可以显著提高蛋白质的乳化活力，水解产物在酸性范围具有相对恒定且最高乳化活力指标，在碱性范围则表现出较低的乳化活力；随着DH的增加，最高乳化活力指标表现的pH范围更宽广，但都保持在112m~2／g以上。复合酶对乳化稳定性有降低趋势，在酸性范围比碱性范围具有相对较高的乳化稳定性；同时，较低DH的水解产物比较高DH的具有相对较低的抗热处理能力。蛋白质浓度为1％~5％时，FP随浓度的增加而增加；泡沫形成后的1min内，1％蛋白质浓度的泡沫较3％和5％的稳定，而15min后，泡沫的稳定性随蛋白质浓度的增加而增加。浓度为3％、DH为19.9％的水解产物，pH4.0时具有最低的起泡性能，而pH10.0时具有最高的起泡能力；pH4.0和8.0是泡沫最不稳定的pH点，而pH10.0时泡沫最稳定。
In this paper, studies on preparation of hydrolysate by use of controlled complicated-enzymes hydrolysis of Harengula Zunasi Bleeker proteins, food functionality and anti- osteoporosis action were carried out.
The studies on material protein peculiarity and its protein fraction showed that the protein of Harengula Zunasi Bleeker can be used in manufacturing functional food, gelatin, bioactive peptides and sea fresh condiment except for fish filling because the ratios of the content of taste amino acids and the content of hydrophobic amino acids to the total content of amino acids in protein were 44.0% and 41.9%, respectively, and that the ratios of the content of stroma protein and the content of myofibrilar protein to the total protein were 52.46% and 4. 19%, respectively. Meanwhile the molecule size of protein fraction was myogen> stroma protein> alkali-soluble protein > myofibrilar protein.
In order to establish the process of the controlled-complicated enzymes hydrolysis, first the optimum conditions of papain, flavorease and complicated enzymes were established by orthogonal trial with the constant time of 2 hours, secondly, the obtained hydrolysates by them at varying hydrolysis time were studied. The results showed that the usage of the complicated enzymes obtained by papain and flavorease may be decreased 22 times than its pre-enzyme and its optimum pH approaches to the pH of fish meat dissolved in water; that the hydrolysis course for the complicated enzymes is described with 3 linear relationship being expressed as follows: DH=0. 0261t+8. 8486(t = 10-210min, r=0. 9834) , MLPL=-0. 0241H + 11. 5616 ( t≠0, r=-0. 9369) and TFAA=0. 1558t+38. 76 (t = 30-210min, r=0. 9891) ; and the complicated enzymes has the strongest hydrolysis specificity toward the peptide linkages supplied by cysteine, phenylalanine and tyrosine and it can break the bond supplied by tryptophane, but papain and flavorease can not.
The kinetic model and relative parameter of
引文
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