菠萝蛋白酶水解泥鳅蛋白制备ACE抑制肽的研究

食品科学 ›› 2012, Vol. 33 ›› Issue (1): 180-185.

菠萝蛋白酶水解泥鳅蛋白制备ACE抑制肽的研究

姚东瑞，盘赛昆，周鸣谦，王淑军，胡金玲

淮海工学院食品工程学院

收稿日期:2011-05-05 修回日期:2012-01-08 出版日期:2012-01-15 发布日期:2012-01-12
通讯作者: 盘赛昆 E-mail:pskgx@163.com
基金资助:
国家“十一五”科技支撑计划项目;江苏省农业科技自主创新基金项目;连云港市科技基础设施建设计划项目

Preparation of ACE Inhibitory Peptides by Bromelain Hydrolysis of Loach (Misgurnus anguillicaudatus) Protein

Received:2011-05-05 Revised:2012-01-08 Online:2012-01-15 Published:2012-01-12

摘要/Abstract

摘要： 为了探讨利用泥鳅蛋白制备功能性肽的可能性，采用高效液相色谱法测定泥鳅肉水解物对血管紧张素转换酶(ACE)的抑制作用，从胰蛋白酶、胃蛋白酶、菠萝蛋白酶、复合风味蛋白酶、复合蛋白酶5种酶中筛选出菠萝蛋白酶作为酶解泥鳅肉制备具有降血压活性水解物的适宜水解酶。在单因素试验的基础上，采用L9(34)正交试验设计对该酶的酶解条件进行优化。结果表明最佳水解条件为：温度55℃，固液比1:3，pH6.5，加酶量1000U/g pro，水解时间90min。在该条件下，水解物的ACE抑制率IC50值为0.0184mg/mL，ACE抑制肽的相对分子质量主要集中在924左右。

关键词: 泥鳅, 酶解, 降血压, ACE, 菠萝蛋白酶

Abstract: The present deals with the enzymatic preparation of angiotensin-converting enzyme (ACE) inhibitory active peptides from loach (Misgurnus anguillicaudatus) protein. An L9(34) orthogonal array design methods based on single factor experiments was used to optimize the hydrolysis conditions for achieving maximum ACE inhibitory activity of loach protein hydrolysates. HPLC was used to determine ACE inhibitory activity of loach protein hydrolysates. Bromelain was found to be more suitable to prepare ACE inhibitory active peptides than trypsin, pepsin, flavourzyme and protamex. The optimal hydrolysis conditions were temperature of 55 ℃, solid-to-liquid ratio of 1:3, pH of 6.5, bromelain dose of 1000 U/g protein and hydrolysis duration of 90 min. The IC50 of the loach protein hydrolysate obtained under these conditions was 0.0184 mg/mL and the relative molecular weight distribution of ACE inhibitory peptides in it was mainly concentrated around 924.

Key words: Misgurnus anguillicaudatus, enzymatic hydrolysis, antihypertensive activity, angiotensin I-converting enzyme (ACE), bromelain

中图分类号:

Q819

姚东瑞，盘赛昆，周鸣谦，王淑军，胡金玲. 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.

参考文献

[1] OTTE J., SHALABY S.M.A., ZAKORA M., et al. Fractionation and identification of ACE-inhibitory peptides from a-lactalbumin and b-casein produced by thermolysin-catalysed hydrolysis[J]. International Dairy Journal 2007, 17: 1460-1472.
[2] 王金玲, 李恒星, 董晓美, 等. 酶解法生产降血压肽的水产蛋白原料筛选研究[J]. 中国食品学报, 2009, 9(3): 111-116.
[3] 王瑛瑶, 王璋,陈尚卫. 花生ACE 抑制活性肽的分离纯化与结构鉴定[J]. 食品科学, 2008, 29(10): 341-344.
[4] RAGHAVAN S.,KRISTINSSON H.G. ACE-inhibitory activity of tilapia protein hydrolysates[J]. Food Chemistry, 2009, 117: 582-588.
[5] CHIANG W.-D., TSOU M.-J., Tsai Z.-Y., et al. Angiotensin I-converting enzyme inhibitor derived from soy protein hydrolysate and produced by using membrane reactor[J]. Food Chemistry, 2006, 98: 725-732.
[6] WU H., HE H., CHEN，X., et al. Purification and identification of novel angiotensin-I-converting enzyme inhibitory peptides from shark meat hydrolysate[J]. Process Biochemistry, 2008, 43: 457-461.
[7] 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: 302-308.
[8] LEE S.H., QIAN Z.J., KIM S.K. A novel angiotensin I converting enzyme inhibitory peptide from tuna frame protein hydrolysate and its antihypertensive effect in spontaneously hypertensive rats[J]. Food Chemistry, 2010, 118: 96-102.
[9] 钦传光, 黄开勋,徐辉碧. 泥鳅中的生物活性分子及其药理作用[J]. 中国生化药物杂志, 2002, 23(1): 47-48.
[10] YOU L., ZHAO M., CUI C., at al. Effect of degree of hydrolysis on the antioxidant activity of loach (Misgurnus anguillicaudatus) protein hydrolysates[J]. Innovative Food Science and Emerging Technologies, 2009 10: 235-240.
[11] 凌先. 泥鳅的食用价值与药用价值[J]. 种养门诊, 2009, 16(5): 116.
[12] 赵振山, 高贵琴, 印杰, 等. 泥鳅和大鳞副泥鳅营养成分分析[J]. 水利渔业, 1999, 19(2): 16-17.
[13] 刘孝华. 泥鳅的生物学特性及养殖技术[J]. 湖北农业科学, 2008, 47(1): 93-95.
[14] GUO, Y., PAN D.,Tanokura M. Optimisation of hydrolysis conditions for the production of the angiotensin-I converting enzyme (ACE) inhibitory peptides from whey protein using response surface methodology[J]. Food Chemistry, 2009, 114: 328-333.
[15] 诺维信(中国)生物技术有限公司, 张家港市金源生物化工有限公司, 中国食品发酵工业研究所, 等. 蛋白酶制剂[S]. 北京: 中国标准化出版社, 2009.
[16] 卫生部食品卫生监督检验所. GB 5009.5-2010 食品中蛋白质的测定[S]. 北京: 中国标准出版社, 2010.
[17] 候曼玲. 食品分析[M]. 北京: 化学工业出版社, 2004: 128-129.
[18] 刘宁, 仇农学, 朱振宝, 等. 杏仁蛋白水解物对血管紧张素转化酶抑制作用的研究[J]. 食品科学, 2009, 30(5): 249-252.
[19] 刘宁,仇农学. 碱性蛋白酶Alcalase水解杏仁蛋白制备ACE抑制肽[J]. 农产品加工?学刊, 2009, (166): 149-152.
[20] 朱迎春, 许小琴,马俪珍. 鲶鱼骨酶解物的降血压肽活性研究[J]. 青岛农业大学学报(自然科学版) 2009, 26(1): 61-65.
[21] 徐娟,吕嘉枥. 乳蛋白水解液中多肽含量测定方法的研究[J]. 食品科技, 2010, 35(12): 275-277.
[22] 盘赛昆, 顾小红, 汤坚, 等. 鲤鱼肉酶解物清除羟自由基的研究[J]. 食品研究与开发, 2009, 30(9): 23-27.
[23] 姚东瑞, 王淑军, 李圆圆, 等. 泥鳅肉酶解物对羟自由基的清除作用[J]. 食品科学, 2010, 31(21): 29-34.
[24] 许庆陵,曾庆祝. 血管紧张素转换酶(ACE)抑制肽的酶法制备[J]. 广州大学学报(自然科学版), 2007, 6(6): 41-44.
[25] 黄艳春, 熊善柏, 赵思明, 等. 鲢肉酶解工艺及其产物对大鼠ACE抑制活性的研究[J]. 食品科学, 2006, 27(1): 203-206.
[26] 吴建平,丁霄霖. 大豆降血压肽的研制(I)--生产高活性ACE I肽酶系的筛选[J]. 中国油脂, 1998, 23(2): 49-51.
[27] MIGUEL M., CONTRERAS M.M., RECIO I., 等. ACE-inhibitory and antihypertensive properties of a bovine casein hydrolysate[J]. Food Chemistry, 2009, 112: 211-214.
[28] NAKAJIMA, K., YOSHIE-STARK Y., OGUSHI M. Comparison of ACE inhibitory and DPPH radical scavenging activities of fish muscle hydrolysates[J]. Food Chemistry, 2009, 114: 844-851.