响应面法优化猪血红蛋白制备ACE抑制肽的酶解工艺

食品科学 ›› 2012, Vol. 33 ›› Issue (23): 209-214.

响应面法优化猪血红蛋白制备ACE抑制肽的酶解工艺

郑炯1,2，邓惠玲1，阚建全1,2,*

1.西南大学食品科学学院 2.重庆市特色食品工程技术研究中心

收稿日期:2011-09-24 修回日期:2012-10-31 出版日期:2012-12-15 发布日期:2012-12-12
通讯作者: 郑炯 E-mail:zhengjiong248@163.com
基金资助:
西南大学基本科研业务费专项资金资助项目

Optimization of Enzymatic Hydrolysis Conditions for ACE Inhibitory Peptides Preparation from Porcine Hemoglobin by Response Surface Methodology

, ,

Received:2011-09-24 Revised:2012-10-31 Online:2012-12-15 Published:2012-12-12

摘要/Abstract

摘要： 以猪血红蛋白为原料，采用胃蛋白酶水解猪血红蛋白制备ACE抑制肽。以体外ACE抑制率和水解度为指标，通过单因素试验对酶解温度、酶解pH值、底物质量浓度、加酶量、酶解时间等酶解工艺参数进行研究，并用响应面法优化酶解工艺，建立二次多项数学模型。结果表明，胃蛋白酶水解猪血红蛋白制备ACE抑制肽的最佳工艺参数为酶解温度37.60℃、酶解pH 1.98、底物质量浓度4.98g/100mL、加酶量3.04%、酶解时间4h，酶解产物的最大ACE抑制率为70.09%。

关键词: 猪血红蛋白, ACE抑制肽, 酶解工艺, 响应面法

Abstract: The enzymatic preparation of angiotensin I-converting enzyme (ACE) inhibitory peptides from porcine hemoglobin using pepsin was optimized by response surface methodology. ACE inhibitory rate in vitro and degree of hydrolysis (DH) were investigated with respect to hydrolysis parameters including temperature, pH, substrate concentration, enzyme dosage and time. As a result, a quadratic polynomial model was developed. The optimum conditions for enzymatic preparation of ACE inhibitory peptides from porcine hemoglobin were found to be hydrolysis at pH 1.98 and 37.60 ℃ for 4 h with a substrate concentration of 4.98 g/100 mL and a enzyme dosage of 3.04%. Under these conditions, the prepared peptides showed an ACE inhibitory rate of 70.09%.

Key words: porcine hemoglobin, ACE inhibitory peptide, enzymatic hydrolysis, response surface methodology

中图分类号:

郑?炯1,2，邓惠玲1，阚建全1,2,*--. 响应面法优化猪血红蛋白制备ACE抑制肽的酶解工艺[J]. 食品科学, 2012, 33(23): 209-214.

参考文献

[1]刘力生, 王文, 姚崇华.中国高血压防治指南[J].中华高血压杂志,2010,18(1):11-29
[2] SKEGGS L,KAHN J,SHUMWAY N.The preparation and function of the angiotensin-I converting enzyme[J].Exp Med.1975,69:295-299
[3]CUSHMAN D,ONDETTI M.Design of potent competitive inhibition of angiotensin converting enzyme[J].Biochemistry,1977,16(4):5484-5488
[4] ATKINSON A, ROBERTSON J. Captopril in the treatment of clinical hypertension and cardiac failure[J]. Lancet.1979;2:836–9
[5]FERREIRA S,tBARTELT D,GREENE L.Isolation of bradykininpotentiating peptides from Bothrops jararaca venom[J].Biochemistry,1970,9:2583-2593
[6]SEKI E.nullOSAJIMA K.Quantitative analysis of digestion resistant ACE inhibitory peptides by small intestinal mucosa[J].. Nipon Skokukin Kagaku Kogaku Kaishi,1996,:-
[7]SHIN Z,Yu R,Park S,et a1.His-His-Leu,an angiotensin I converting enzyme inhibitory peptide derived from Korean Soybean Paste, exerts antihypertensive activity in vivo[J].Journal of Agricultural and Chemistry,2001,49:3004-3009
[8]管骁,刘静,王立等.高活性燕麦蛋白源ACE抑制肽的制备、纯化及结构鉴定[J].高等学校化学学报,2009,30,10:1992-1997
[9]WANG X,CHEN P,LIANG S.Liquid chromatography tandem mass spectrometry analysis of a synthestic nonapeptide and its main by products[J].Chinese Journal of Biochemistry and Molecular Biology,2003,19,5:576-581
[10]黄家音,朱宇洁,沈金玉.玉米蛋白制备降血压肽的研究[J].食品科学,2007,28,12:290-293
[11] MARUYAMA S,MITACH H,TANAKA H,et al. Studies on the active site and antihypertensive activity of angiotensin I–converting enzyme inhibitors derived from casein[J]. Agric Biol Chem,1987a,51:1581-1586
[12]KUNIO S,JIUN R.Identification of Antihypertensive Peptides from Peptic Digest of Two Microalgae,Chlorella vulgaris and Spirulina platensis[J].Marine Biotechnology,2001,3:305-314
[13]FUJITA.LKPNM: a prodrugtype ACE-inhibitory peptide derived from fish protein[J].Immunopharmacology,1999,44:123-
[14]FAHMI A,MORIMURA S,GUO H,et al.Production of angiotensin converting enzyme inhibitory peptides from sea bream scales[J].Process Biochem,2004,39:1195-2000
[15]LEE J,KWON D,SHIN H,et al.Purification and identification angiotensin-I-converting enzyme inhibitory peptides from kidney bean protein hydrolysate[J].Food Science and Biotechnology,1999,8:172-178
[16]MEANO M,YAMAMOTO N,TAKANO T.Identification of an antihypertensive peptide from casein hydrolysate produced by a proteinase from Lactobacillus helveticus CP790[J].Jour Dairy Sci,1996,79:1316-1321
[17]KAZUE M.nullMIKIO F,MASAYOSHI K ,et al. Antihypertensive effect of angiontensin I-converting enzyme inhibitory peptides derived from hemoglobin[J].. European Journal of Pharmacology,1996,:-
[18] YU Y,HU J,BAI X,et al. Preparation and function of oligopeptide-enriched hydrolysate from globin by pepsin.Process Biochem 2006:41(7):1589–93
[19]胡鑫,罗永康.猪血红蛋白酶解制备ＡＣＥ抑制肽的研究[J].肉类研究,2009,(12):21-24
[20]Zhao Qiuyu,Sannier F,Piot J M.Kinetics of appearance of four hemorphins from bovine hemoglobin peptic hydrolysates by HPLC coupled with photodiode array detection[J].Biochimica et Biophysica Acta,1996,1295:73-80
[21]余勃,陆兆新.微生物发酵法产大豆多肽液水解度的测定[J].食品科学,2005,26,4:104-107
[22]WU J,ALUKO R, MUIR A.Imp roved Method for Direct High-performance Liquid Chromatography Assay of Angiotensin-converting Enzyme-catalyzed Reaction[J].of Chromatogr A,2002,950:125-131
[23]ZHAO Y, LI B, LIU Z, et al.Antihypertensive effect and purification of an ACE inhibitory peptide from sea cucumber gelatin hydrolysate[J].Process Biochemistry,2007,42(12):1586-1591