Study on enzymatic hydrolysis process of fish collagen and antihypertensive peptides of collagen
-
摘要: 本研究通过单因素实验和正交实验优化胃蛋白酶水解鱼胶原蛋白制备胶原多肽的工艺。采用高效液相色谱法测定海水鱼与淡水鱼胶原多肽血管紧张素转换酶(ACE)的抑制活性来表征其降血压功能。研究表明:胃蛋白酶制备鱼胶原多肽的最佳酶解工艺条件是水解温度为37℃,初始p H为2.5,酶与底物比例为1∶125(w/w),底物质量分数为3%(w/v),水解时间为12 h,此时鱼胶原蛋白溶液的水解度为19.17%。研究发现海洋鱼胶原蛋白和淡水鱼胶原蛋白的酶解上清液均具有较高的ACE抑制活性,半抑制浓度IC50分别为1.34 mg/m L和1.52 mg/m L。利用超滤离心管获得的不同分子量组分中,分子量小于3 ku的海洋胶原多肽CP-Ⅰ和淡水胶原多肽CP-Ⅲ的ACE抑制活性的半抑制浓度IC50分别为0.43 mg/m L和0.60 mg/m L,而相应分子量大于3 ku的海洋胶原多肽CP-Ⅱ和淡水胶原多肽CP-Ⅳ的半抑制浓度IC50分别为1.78 mg/m L和1.69 mg/m L。Abstract: Single-factor and orthogonal experiments were employed to optimize the preparation process of collagen polypeptide through the hydrolysis of the fish collagen by pepsin.The angiotensin converting enzyme ( ACE) inhibitory activity of marine and freshwater fish collagen was determined by high performance liquid chromatography to characterize its blood pressure-lowering function.It showed that the optimal hydrolysis condition of pepsin was emzyme-to-substrate ratio of 1 ∶ 125, substrate concentration 3%, temperature 37 ℃, initial p H2.5 and reaction time 12 h while DH was 19.17%.The results showed that the supernatant of enzymatic hydrolysis of marine fish collagen and freshwater fish collagen possessed higher ACE inhibitory activity, while the IC50 of their hydrolysates to ACE were 1.34 mg/m L and 1.52 mg/m L. The hydrolysate was separated by Amicon Ultra-4 Centrifugal Filter Devices with the molecular weight lower than 3 ku. The inhibitory activities of the two fractions was determined.The results showed that the IC50 of CP-Ⅰ ( ≤3 ku) and CP-Ⅲ ( ≤3 ku) were 0.43 mg/m L and0.60 mg/m L. While the IC50 of CP-Ⅱ ( ≥3 ku) and CP-Ⅳ ( ≥3 ku) were 1.78 mg/m L and 1.69 mg/m L.
-
Keywords:
- angiotensin converting enzyme /
- fish /
- collagen protein /
- bioactive peptide
-
[1] 崔和平, 郭兴凤.食源性降血压肽制备研究进展[J].粮食与油脂.2013, 26 (02) :05-08. [2] 梁汉萦, 刘成梅, 刘伟.食源性ACE抑制肽的研究进展[J].食品研究与开发.2007, 28 (08) :156-158. [3] Guan-Hong Lia, Guo-Wei Lea, Yong-Hui Shi, et al.Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects.Nutrition Research, 2004, 24 (7) :469-486.
[4] 洪伟, 薛正莲, 陈玲.酪蛋白制备ACE抑制肽的酶解工艺优化[J].食品与发酵科技.2010, 46 (02) :37-40. [5] 杨湘华, 李江.食源性降血压肽构效关系与制备研究进展[J].食品研究与开发.2016, 37 (03) :217-220. [6] 刘赛赛, 伍善广, 冯学珍, 等.木瓜蛋白酶水解酪蛋白制备降血压肽的工艺优化[J].食品科技, 2014, 39 (10) :48-52. [7] 吴红洋, 姜太玲, 申光辉, 等.花椒籽蛋白降血压肽的加工适应性研究[J].中国油脂, 2015, 40 (2) :24-29. [8] 杨英士, 陈伟, 杨海燕, 等.昆仑雪菊中血管紧张素转化酶活性抑制成分的分离鉴定[J].南京农业大学学报, 2015, 38 (1) :146-151. [9] Saleh AS M, Zhang Q, Shen Q.Recent research in antihypertensive activity of food protein-derived hydrolyzates and peptides[J].Critical Reviews in Food Science and Nutrition, 2016, 56 (5) :760-787.
[10] Shu Hua Xia, Zhang Wang, Shi Ying Xu.Characteristics of Bellamya purificata snail foot protein and enzymatic hydrolysates[J].Food Chemistry, 2007, 101 (3) :1188-1196.
[11] Park Y W, Nam M S.Bioactive Peptides in Milk and Dairy Products:A Review[J].Korean journal for food science of animal resources, 2015, 35 (6) :831.
[12] Mirdhayati I, Hermanianto J, Wijaya C H, et al.Angiotensin converting enzyme (ACE) inhibitory and antihypertensive activities of protein hydrolysate from meat of Kacang goat (Capra aegagrus hircus) [J].Journal of the Science of Food and Agriculture, 2016.
[13] Cheung Randy Chi Fai, Ng Tzi Bun, Wong Jack Ho.Marine Peptides:Bioactivities and Applications[J].Marine Drugs, 2015, 13 (7) :4006-4043.
[14] Liu X, Zhang M, Shi Y, et al.Production of the angiotensin I converting enzyme inhibitory peptides and isolation of four novel peptides from jellyfish (Rhopilema esculentum) protein hydrolysate[J].Journal of the Science of Food and Agriculture, 2015.
[15] García-Moreno P J, Espejo-Carpio F J, Guadix A, et al.Production and identification of angiotensin I-converting enzyme (ACE) inhibitory peptides from Mediterranean fish discards[J].Journal of Functional Foods, 2015, 18:95-105.
[16] 吴靖娜, 许永安, 王茵, 等.罗非鱼鱼皮胶降血压肽的初步分离及性质研究[J].食品工业, 2012, 11:032. [17] 杨丽, 林福兰.食品检验与分析实验[M].广州:华南理工大学出版社, 2013. [18] Jianping Wu, Rotimi E Aluko, Alister D Muir.Improved method for direct high-performance liquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions.Journal of Chromatography A, 2002, 950 (1) :125-130.
[19] Xin Rui, Joyce I.Boye, Benjamin K.Simpson, Shiv O.Prasher.Purification and characterization of angiotensin I-converting enzyme inhibitory peptides of small red bean (Phaseolus vulgaris) hydrolysates[J].Journal of Functional Foods, 2013, 5 (3) :1116-1124.
[20] Rui-Zeng Gu, Chen-Yue Li, Wen-Ying Liu, Wei-Xue Yi, Mu-Yi Cai.Angiotensin I-converting enzyme inhibitory activity of low-molecular-weight peptides from Atlantic salmon (Salmo salar L.) skin.Food Research International 2011, 44 (5) :1536-1540.
计量
- 文章访问数:
- HTML全文浏览量:
- PDF下载量: