青鳞鱼蛋白酶解工艺及产物组分分析
|
摘要
以青鳞鱼鱼肉为原料,用风味蛋白酶对其进行水解处理制取水解蛋白质,通过单因素试验考察了酶解温度、加酶量、酶解时间、底物浓度、pH值等不同因素对青鳞鱼蛋白质水解物中游离氨基态氮含量和水解度的影响。在此基础上,采用Box-Behnken试验设计,优化了青鳞鱼水解蛋白质的制备工艺。结果表明:在水解温度50℃、pH值6.5、底物∶水=1.0∶1.9(质量比)、加酶量2 200 U/g、酶解时间300 min条件下,青鳞鱼蛋白质的水解度为14.82%,与理论预测值之间误差仅为0.29%,游离氨基态氮含量为1.946 mg/ml,表明采用响应面法优化得到的青鳞鱼蛋白质酶解工艺参数准确可靠。酶解产物主要由3～19肽构成,占总酶解产物的71.63%,其中2～4肽占总酶解产物的48.86%。表明青鳞鱼鱼肉经风味蛋白酶水解时主要生成聚合度为2～4的小分子寡肽,风味蛋白酶对青鳞鱼蛋白质具有较强水解作用。
Using Harengula zunast as raw material, the hydrolyzed protein was prepared by hydrolysis of fish meat with flavourzyme. The effects of hydrolysis temperature, enzyme dosage, enzymolysis time, substrate concentration and pH value on the content and hydrolysis degree of free amino nitrogen in hydrolysate of Harengula zunast protein were investigated by single-factor experiments. On this basis, the preparation process of hydrolyzed protein was optimized by Box-Behnken experimental design. The optimum hydrolysis parameters were as follows: temperature 50 ℃, pH 6.5, mass ratio of substrate concentration 1.0∶1.9, enzyme dosage 2 200 U/g, and reaction time 300 min. Under the optimized technology, the hydrolysis degree of Harengula zunast protein was 14.82%, and the relative error between predicted value and actual value was 0.29%, the content of free amino nitrogen was 1.946 mg/ml. The results showed that the parameters of protease hydrolysate obtained by response surface method were accurate and reliable. The enzymatic hydrolysates were mainly composed of 3-19 peptides, accounting for 71.63% of the total hydrolysates, and the 2-4 peptide accounted for 48.86%. These results indicanted that the small molecular oligopeptides with the degree of polymerization of 2-4 were mainly produced when Harengula zunast fish meat was hydrolyzed by flavourzyme, and flavourzyme had strong hydrolysis effect on Harengula zunast protein.
Using Harengula zunast as raw material, the hydrolyzed protein was prepared by hydrolysis of fish meat with flavourzyme. The effects of hydrolysis temperature, enzyme dosage, enzymolysis time, substrate concentration and pH value on the content and hydrolysis degree of free amino nitrogen in hydrolysate of Harengula zunast protein were investigated by single-factor experiments. On this basis, the preparation process of hydrolyzed protein was optimized by Box-Behnken experimental design. The optimum hydrolysis parameters were as follows: temperature 50 ℃, pH 6.5, mass ratio of substrate concentration 1.0∶1.9, enzyme dosage 2 200 U/g, and reaction time 300 min. Under the optimized technology, the hydrolysis degree of Harengula zunast protein was 14.82%, and the relative error between predicted value and actual value was 0.29%, the content of free amino nitrogen was 1.946 mg/ml. The results showed that the parameters of protease hydrolysate obtained by response surface method were accurate and reliable. The enzymatic hydrolysates were mainly composed of 3-19 peptides, accounting for 71.63% of the total hydrolysates, and the 2-4 peptide accounted for 48.86%. These results indicanted that the small molecular oligopeptides with the degree of polymerization of 2-4 were mainly produced when Harengula zunast fish meat was hydrolyzed by flavourzyme, and flavourzyme had strong hydrolysis effect on Harengula zunast protein.
引文
[1] 王杏珠.青鳞鱼加工和利用的研究[J].水产科技情报,1995,16(1):28-32.
[2] 邓尚贵,杨萍.青鳞鱼肌肉蛋白复合酶水解机制研究[J].食品科学,2004,25(3):67-75.
[3] 范蕴莹.不同种类低值鱼酶解效果的比较及优化[J].现代食品科技,2012,28(7):814-818.
[4] 段蕊,张俊杰.海洋低值小杂鱼的加工方向和现状[J].食品与发酵工业,2000,26(6):53-56.
[5] 张海莲.浅谈海洋低值小杂鱼的综合利用[J].河北渔业,2005 (4):7-8.
[6] 师晓栋,何海伦,王运涛,等.酶法进行海洋低值蛋白资源高值化利用初探[J].海洋科学,2001,25(3):4-7.
[7] 杨萍,邓尚贵,吴玉廉.蛋白酶及其作用方式对制取青鳞鱼肉水解蛋白的影响[J].食品与发酵工业,2003,29(4):91-94.
[8] 杨萍,邓尚贵.木瓜蛋白酶在制取青鳞鱼可溶性蛋白中的应用[J].广东海洋大学学报,2002,22(3):38-41.
[9] DENG S G,YANG P,YANG Z J,et al.Conditions and hydrolysate of the enzymolysis of harengula zunasi B.muscle proteins by flavorease[J].Journal of Zhanjiang Ocean University,2004,24 (1):20-27.
[10] DENG S G,PENG Z Y,CHEN F,et al.Amino acid composition and anti-anaemia action of hydrolyzed offal protein from Harengula Zunasi,Bleeker[J].Food Chemistry,2004,87(1):97-102.
[11] 常志娟,邓尚贵.青鳞鱼水解蛋白复合果味酸奶的研制[J].饮料工业,2008,11(2):35-38.
[12] KAWAGUTI S,KAMISHIMA Y.Electron microscopy on the blue back of a clupeoid fish,harengula zunasi[J].Proceedings of the Japan Academy,2006,42(4):389-393.
[13] 曹文红,冯思涛,莫敏婷,等.青鳞鱼酶解工艺正交优化及其产物分子量分布的研究[J].食品科技,2013(4):130-135.
[14] 郭晶晶,马海乐,程宇,等.风味蛋白酶水解草鱼肉的试验研究[J].食品工业科技,2014,35(13):248-252.
[15] 邵俊杰,朱昱璇,黄鸿兵,等.不同蛋白酶酶解河蚬蛋白的比较[J].江苏农业学报,2017,33(4):921-926.
[16] 候曼玲.食品分析[M].北京:化学工业出版社,2004:76-77.
[17] WALSH D J,CLEARY D,MCCARTHY E,et al.Modification of the nitrogen solubility properties of soy protein isolate following proteolysis and transglutaminase cross-linking[J].Food Research International,2003,36(7):677-683.
[18] 陈杰,马美湖.风味蛋白酶水解蛋清工艺条件的研究[J].现代食品科技,2007,23(7):43-44.
[19] 刘爽,夏杨毅,尚永彪.鸡蛋蛋清蛋白酶解产物的研究进展[J].食品科学,2013,34(15):409-414.
[20] 汪怀建,谭文津,丁雪杉,等.超声波辅助提取玉米芯中木聚糖条件优化研究[J].中国粮油学报,2009,24(7):50-54.
[21] 陈爱瑛,贾建萍,叶小弟,等.鱼皮胶原蛋白水解液中小分子寡肽的分子量分布测定[J].中国现代应用药学,2011,28(2):160-162.
[22] 张俊杰,段蕊.高蛋白鱼粉的加工[J].淮海工学院学报(然科学版),2000,9 (2):44-46.
[23] 吉挺,张焕新,殷玲.油菜蜂花粉多肽饮料的研制及抗氧化活性评价[J].食品与发酵工业,2016,42(8):135-141.
[24] 常志娟.采用复合酶水解青鳞鱼蛋白及水解产物特性研究[D].湛江:广东海洋大学,2005.
[25] 李莉,张赛,何强,等.响应面法在试验设计与优化中的应用[J].实验室研究与探索,2015,34(8):41-45.
[26] KUMAR A,PRASAD B,MISHRA I M.Optimization of process parameters for acrylonitrile removal by a low-cost adsorbent using Box-Behnken design.[J].Journal of Hazardous Materials,2008,150(1):174-182.
[27] 姚东瑞,盘赛昆,周鸣谦,等.菠萝蛋白酶水解泥鳅蛋白制备ACE抑制肽的研究[J].食品科学,2012,33(1):180-185.
[28] 马永强,赵毅.大豆蛋白水解物中肽分子分布的研究[J].中国粮油学报,2001,16(3):15-17.
[29] 刘文斌,尹君,王恬.棉粕蛋白及其枯草蛋白酶酶解产物的组分分析[J].食品与发酵工业,2005,31(11):105-108.
[30] WANG J,HU J,CUI J,et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats[J].Food Chemistry,2008,111(2):302-308.
[2] 邓尚贵,杨萍.青鳞鱼肌肉蛋白复合酶水解机制研究[J].食品科学,2004,25(3):67-75.
[3] 范蕴莹.不同种类低值鱼酶解效果的比较及优化[J].现代食品科技,2012,28(7):814-818.
[4] 段蕊,张俊杰.海洋低值小杂鱼的加工方向和现状[J].食品与发酵工业,2000,26(6):53-56.
[5] 张海莲.浅谈海洋低值小杂鱼的综合利用[J].河北渔业,2005 (4):7-8.
[6] 师晓栋,何海伦,王运涛,等.酶法进行海洋低值蛋白资源高值化利用初探[J].海洋科学,2001,25(3):4-7.
[7] 杨萍,邓尚贵,吴玉廉.蛋白酶及其作用方式对制取青鳞鱼肉水解蛋白的影响[J].食品与发酵工业,2003,29(4):91-94.
[8] 杨萍,邓尚贵.木瓜蛋白酶在制取青鳞鱼可溶性蛋白中的应用[J].广东海洋大学学报,2002,22(3):38-41.
[9] DENG S G,YANG P,YANG Z J,et al.Conditions and hydrolysate of the enzymolysis of harengula zunasi B.muscle proteins by flavorease[J].Journal of Zhanjiang Ocean University,2004,24 (1):20-27.
[10] DENG S G,PENG Z Y,CHEN F,et al.Amino acid composition and anti-anaemia action of hydrolyzed offal protein from Harengula Zunasi,Bleeker[J].Food Chemistry,2004,87(1):97-102.
[11] 常志娟,邓尚贵.青鳞鱼水解蛋白复合果味酸奶的研制[J].饮料工业,2008,11(2):35-38.
[12] KAWAGUTI S,KAMISHIMA Y.Electron microscopy on the blue back of a clupeoid fish,harengula zunasi[J].Proceedings of the Japan Academy,2006,42(4):389-393.
[13] 曹文红,冯思涛,莫敏婷,等.青鳞鱼酶解工艺正交优化及其产物分子量分布的研究[J].食品科技,2013(4):130-135.
[14] 郭晶晶,马海乐,程宇,等.风味蛋白酶水解草鱼肉的试验研究[J].食品工业科技,2014,35(13):248-252.
[15] 邵俊杰,朱昱璇,黄鸿兵,等.不同蛋白酶酶解河蚬蛋白的比较[J].江苏农业学报,2017,33(4):921-926.
[16] 候曼玲.食品分析[M].北京:化学工业出版社,2004:76-77.
[17] WALSH D J,CLEARY D,MCCARTHY E,et al.Modification of the nitrogen solubility properties of soy protein isolate following proteolysis and transglutaminase cross-linking[J].Food Research International,2003,36(7):677-683.
[18] 陈杰,马美湖.风味蛋白酶水解蛋清工艺条件的研究[J].现代食品科技,2007,23(7):43-44.
[19] 刘爽,夏杨毅,尚永彪.鸡蛋蛋清蛋白酶解产物的研究进展[J].食品科学,2013,34(15):409-414.
[20] 汪怀建,谭文津,丁雪杉,等.超声波辅助提取玉米芯中木聚糖条件优化研究[J].中国粮油学报,2009,24(7):50-54.
[21] 陈爱瑛,贾建萍,叶小弟,等.鱼皮胶原蛋白水解液中小分子寡肽的分子量分布测定[J].中国现代应用药学,2011,28(2):160-162.
[22] 张俊杰,段蕊.高蛋白鱼粉的加工[J].淮海工学院学报(然科学版),2000,9 (2):44-46.
[23] 吉挺,张焕新,殷玲.油菜蜂花粉多肽饮料的研制及抗氧化活性评价[J].食品与发酵工业,2016,42(8):135-141.
[24] 常志娟.采用复合酶水解青鳞鱼蛋白及水解产物特性研究[D].湛江:广东海洋大学,2005.
[25] 李莉,张赛,何强,等.响应面法在试验设计与优化中的应用[J].实验室研究与探索,2015,34(8):41-45.
[26] KUMAR A,PRASAD B,MISHRA I M.Optimization of process parameters for acrylonitrile removal by a low-cost adsorbent using Box-Behnken design.[J].Journal of Hazardous Materials,2008,150(1):174-182.
[27] 姚东瑞,盘赛昆,周鸣谦,等.菠萝蛋白酶水解泥鳅蛋白制备ACE抑制肽的研究[J].食品科学,2012,33(1):180-185.
[28] 马永强,赵毅.大豆蛋白水解物中肽分子分布的研究[J].中国粮油学报,2001,16(3):15-17.
[29] 刘文斌,尹君,王恬.棉粕蛋白及其枯草蛋白酶酶解产物的组分分析[J].食品与发酵工业,2005,31(11):105-108.
[30] WANG J,HU J,CUI J,et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats[J].Food Chemistry,2008,111(2):302-308.