干酪乳杆菌D400产ACE抑制肽的研究
|
摘要
ACE抑制肽具有降血压功能,采用酶解或微生物发酵是目前获取乳源性ACE抑制肽的主要手段。干酪乳杆菌属于同型发酵乳酸菌,有较好的产酸活性及蛋白水解活力,广泛用于多种乳制品生产上。本文从ACE抑制活性菌株的筛选和鉴定、发酵乳ACE抑制率的影响因素、ACE抑制肽的胃肠耐受性及乳酸菌产ACE抑制肽的机制等方面对干酪乳杆菌的ACE抑制活性进行了研究,旨在为应用干酪乳杆菌到相关食品中提供依据。主要研究结果如下:
1、D400和Y5-2b在经pH=2.5的人工胃液消化4 h后,活菌率仍能达到89.09%和86.28%,而经过人工肠液和0.3%胆盐消化后活菌率则仍能达到96%~98%,均表现为良好的耐胃肠环境性能。D400无论在乳中的生长状态,发酵性能还是蛋白水解活力方面都优于Y5-2b,ACE抑制活性试验证明, D400发酵乳的ACE抑制率高于Y5-2b。因此D400被选作本次研究的试验菌株。基于生理生化和16S rRNA基因序列的试验数据,乳杆菌D400被鉴定为干酪乳杆菌(Lactobacillus casei)。
2、随着L.casei D400细胞生物量和发酵乳滴定酸度的上升,菌株的蛋白水解活力以及对ACE的抑制率随之上升,表明随着菌株生长,含在乳蛋白中的ACE抑制肽逐渐被释放出来,菌株的蛋白水解活力与ACE抑制活性有着明显的相关性。3、单因素试验表明,发酵温度、发酵时间和无脂乳固形物浓度都会影响菌株的
蛋白水解活力,影响其ACE抑制效率。优化后的L.casei D400在乳中的发酵条件是:无脂乳固形物浓度为10%,发酵时间为24 h,发酵温度为37°C,在这样的条件下,L.casei D400的ACE抑制率是81.15%。
4、模拟胃肠环境试验结果表明,经胃蛋白酶和胰蛋白酶的作用后,L.casei D400的ACE抑制率由消化前的81.15%下降到消化后的67.83%,下降程度不是很大,说明L.casei D400产生的ACE抑制肽具有一定的耐受胃肠道内蛋白酶消化能力。
5、对L.casei D400发酵乳中的氨基酸组分与含量分析表明,疏水性氨基酸含量在总氨基酸中的比重也由发酵前的22.29%上升为48.50%,脯氨酸含量在总氨基酸中的比重大幅上升,由发酵前的1.67%上升为18.75%,且相对增加量高于其它氨基酸。亮氨酸和苏氨酸的含量相对增加较多,异亮氨酸、精氨酸、赖氨酸、缬氨酸其次,且新生成了蛋氨酸和苯丙氨酸,结合已知ACE抑制肽的序列结构,符合了产ACE抑制肽的条件。因此,L.casei D400产ACE抑制肽的原因是其具有水解蛋白产生脯氨酸及其他疏水性氨基酸和芳香族氨基酸的能力。
ACE inhibitory peptide has a function on lowering blood pressure. Enzymatic or microbial fermentation is the primary means of obtaining ACE inhibitory peptides from milk. Lactobacillus casei belong to the same type fermentation of lactic acid bacteria, has good acid production activity and protein hydrolysis activity, widely used in the various of dairy products. This paper studies screening and identification of the Lactobacillus casei, the determination of proteolytic activity of the strain, ACE inhibitory activity of the strain and contents of amino acids in fermentation milk whey and other aspects, which establishes screening model of Lactobacillus casei with ACE inhibitory activity. Major findings are as follows:
1. After 4 hours digestion by the artificial gastric juice( pH = 2.5), the viable rate of D400 and Y5-2b can reach 89.09% and 86.28%, and the viable rate of digestion is still 96% to 98% after the artificial intestinal juice and 0.3% concentration bile salt. Therefore, D400 and Y5-2b have good resistance to gastrointestinal environmental performance. Further more, D400 in terms of the growth state in the milk, acid production or proteolytic activity areas were higher than Y5-2b, ACE inhibitory activity of D400 fermented milk was higher than Y5-2b. therefore, selected L.casei D400 as the experimental strains. Based on experiment data of physiological, biochemical and 16S rRNA gene sequence, determins D400 and Y5-2b as Lactobacillus casei.
2. With the cell biomass and titratable acidity of L.casei D400 fermented milk increased, the proteolytic activity and the ACE inhibition rate rised, which indicated with the growth of cell biomass, ACE inhibitory peptide included in the milk protein gradually released, proteolytic activity and ACE inhibition has significant correlation.
3. Single factor test showed that the fermentation temperature, fermentation time and the concentration of non-fat milk solids will affect the proteolytic activity and the efficiency of ACE inhibition of the strain. Optimized L.casei D400 in milk fermentation conditions are: SNF concentration of 10%, fermentation time 24 h, fermentation temperature 37°C. In such conditions, ACE inhibition rate of L. casei D400 is 81.15%.
4. After the digestion of pepsin and trypsin, ACE inhibition rate of L.casei D400 decreased from 81.15% before to 67.83%, indicating ACE inhibitory peptides produced by L.casei D400 has a certain tolerance to the gastrointestinal tract protease digestion.
5. The amino acids content in the L.casei D400 fermented milk analysis show that the hydrophobic amino acids in the proportion of the total amino acids increased from the fermentation before 22.29% to 48.50%, proline proportion in the total amino acid has increased significantly from 1.67% before to 18.75%, and the relative increase contents was higher than other amino acids. Leucine and threonine content were relatively large increase, isoleucine, arginine, lysine, valine followed, and found methionine and phenylalanine. Combination structure of ACE inhibitory peptide sequence which has known, the conditions of producing ACE inhibitory peptides formed. Therefore, the capacity of L.casei D400 produce ACE inhibitory peptides is because of its hydrolyzed protein activity producing proline, other hydrophobic amino acids and aromatic amino acid.
1、D400和Y5-2b在经pH=2.5的人工胃液消化4 h后,活菌率仍能达到89.09%和86.28%,而经过人工肠液和0.3%胆盐消化后活菌率则仍能达到96%~98%,均表现为良好的耐胃肠环境性能。D400无论在乳中的生长状态,发酵性能还是蛋白水解活力方面都优于Y5-2b,ACE抑制活性试验证明, D400发酵乳的ACE抑制率高于Y5-2b。因此D400被选作本次研究的试验菌株。基于生理生化和16S rRNA基因序列的试验数据,乳杆菌D400被鉴定为干酪乳杆菌(Lactobacillus casei)。
2、随着L.casei D400细胞生物量和发酵乳滴定酸度的上升,菌株的蛋白水解活力以及对ACE的抑制率随之上升,表明随着菌株生长,含在乳蛋白中的ACE抑制肽逐渐被释放出来,菌株的蛋白水解活力与ACE抑制活性有着明显的相关性。3、单因素试验表明,发酵温度、发酵时间和无脂乳固形物浓度都会影响菌株的
蛋白水解活力,影响其ACE抑制效率。优化后的L.casei D400在乳中的发酵条件是:无脂乳固形物浓度为10%,发酵时间为24 h,发酵温度为37°C,在这样的条件下,L.casei D400的ACE抑制率是81.15%。
4、模拟胃肠环境试验结果表明,经胃蛋白酶和胰蛋白酶的作用后,L.casei D400的ACE抑制率由消化前的81.15%下降到消化后的67.83%,下降程度不是很大,说明L.casei D400产生的ACE抑制肽具有一定的耐受胃肠道内蛋白酶消化能力。
5、对L.casei D400发酵乳中的氨基酸组分与含量分析表明,疏水性氨基酸含量在总氨基酸中的比重也由发酵前的22.29%上升为48.50%,脯氨酸含量在总氨基酸中的比重大幅上升,由发酵前的1.67%上升为18.75%,且相对增加量高于其它氨基酸。亮氨酸和苏氨酸的含量相对增加较多,异亮氨酸、精氨酸、赖氨酸、缬氨酸其次,且新生成了蛋氨酸和苯丙氨酸,结合已知ACE抑制肽的序列结构,符合了产ACE抑制肽的条件。因此,L.casei D400产ACE抑制肽的原因是其具有水解蛋白产生脯氨酸及其他疏水性氨基酸和芳香族氨基酸的能力。
ACE inhibitory peptide has a function on lowering blood pressure. Enzymatic or microbial fermentation is the primary means of obtaining ACE inhibitory peptides from milk. Lactobacillus casei belong to the same type fermentation of lactic acid bacteria, has good acid production activity and protein hydrolysis activity, widely used in the various of dairy products. This paper studies screening and identification of the Lactobacillus casei, the determination of proteolytic activity of the strain, ACE inhibitory activity of the strain and contents of amino acids in fermentation milk whey and other aspects, which establishes screening model of Lactobacillus casei with ACE inhibitory activity. Major findings are as follows:
1. After 4 hours digestion by the artificial gastric juice( pH = 2.5), the viable rate of D400 and Y5-2b can reach 89.09% and 86.28%, and the viable rate of digestion is still 96% to 98% after the artificial intestinal juice and 0.3% concentration bile salt. Therefore, D400 and Y5-2b have good resistance to gastrointestinal environmental performance. Further more, D400 in terms of the growth state in the milk, acid production or proteolytic activity areas were higher than Y5-2b, ACE inhibitory activity of D400 fermented milk was higher than Y5-2b. therefore, selected L.casei D400 as the experimental strains. Based on experiment data of physiological, biochemical and 16S rRNA gene sequence, determins D400 and Y5-2b as Lactobacillus casei.
2. With the cell biomass and titratable acidity of L.casei D400 fermented milk increased, the proteolytic activity and the ACE inhibition rate rised, which indicated with the growth of cell biomass, ACE inhibitory peptide included in the milk protein gradually released, proteolytic activity and ACE inhibition has significant correlation.
3. Single factor test showed that the fermentation temperature, fermentation time and the concentration of non-fat milk solids will affect the proteolytic activity and the efficiency of ACE inhibition of the strain. Optimized L.casei D400 in milk fermentation conditions are: SNF concentration of 10%, fermentation time 24 h, fermentation temperature 37°C. In such conditions, ACE inhibition rate of L. casei D400 is 81.15%.
4. After the digestion of pepsin and trypsin, ACE inhibition rate of L.casei D400 decreased from 81.15% before to 67.83%, indicating ACE inhibitory peptides produced by L.casei D400 has a certain tolerance to the gastrointestinal tract protease digestion.
5. The amino acids content in the L.casei D400 fermented milk analysis show that the hydrophobic amino acids in the proportion of the total amino acids increased from the fermentation before 22.29% to 48.50%, proline proportion in the total amino acid has increased significantly from 1.67% before to 18.75%, and the relative increase contents was higher than other amino acids. Leucine and threonine content were relatively large increase, isoleucine, arginine, lysine, valine followed, and found methionine and phenylalanine. Combination structure of ACE inhibitory peptide sequence which has known, the conditions of producing ACE inhibitory peptides formed. Therefore, the capacity of L.casei D400 produce ACE inhibitory peptides is because of its hydrolyzed protein activity producing proline, other hydrophobic amino acids and aromatic amino acid.
引文
[1]何海伦,陈秀兰,孙彩云,等.血管紧张素转化酶抑制肽的研究进展[J].中国生物工程杂志,2004, 24(9):7-11.
[2] Julian Tudor Hart,Tom Fahey,Wendy Savage.高血压权威指南[M].石油工业出版社,2001,7.
[3]《中国高血压防治指南2005年修订版》,中国高血压防治指南起草委员会.
[4]郭胜才,冯友根.抗高血压药物研究新进展.中国药学杂志,2001, 36:583-587.
[5]徐贵成,霍保民,王秋风,等.通心络胶囊治疗风湿性心瓣膜病心力衰竭临床观察[J].浙江中西医结合志,2002,12(1):5-7.
[6]安茂竹,孙静,高景芳,等.血管紧张素转换酶抑制剂的不良反应及防治[J].中国新药杂志,1999, 8(5):403-404.
[7]吴建平.大豆降压肽的研制[D].博士学位论文.无锡:江南大学,1998.
[8]靳姝杰,袭荣刚,王晓波.血管紧张素转换酶抑制剂的不良反应和禁忌症[J].药学实践杂志,2000, 18(3):166-168.
[9]赵海珍,陆兆新,刘战民.天然食品来源的血管紧张素转换酶抑制肽的研究进展[J].中国生化药物杂志,2004,25(5):315-317.
[10]辛志宏,马海乐.食品蛋白质中降血压肽的功能与应用[J].食品与发酵工业,2003,29(8),32-36.
[11] Johnston JI,Franz VL.Renin-angiotensin system: a dual tissue and hormonal system for cardiovascular control[J]. J Hypertens,1992,10:S13-S26.
[12] Ferreira SH. Abradykinin-potentiating factor(BPF) present in the venom of Bothrops jararaca. Brit J Pharmacol,1965,24:163-169.
[13] Bakhle Y S.Conversion of angiotensin-Ⅰto angiotensin-Ⅱby cell free extracts of dog lung[J]. Nature 1968, 220 (5170):919-921.
[14] Ondetti M A, Williams N J, SaboEF, et al. Angiotensin-converting enzyme inhibitors from the Venom of Bothrops Jararaca.Isolation, elucidation of structure, and synthesis.[J] Biochemisty 1971,10(22):4033-4039.
[15] Cheung H S, Cushman D W. Inhibition of homogeneous angiotensin-converting enzyme of rabbit lung by synthetic venom peptides of Bothrops Jararaca[J]. Biochim Biophys Acta 1973, 293(2):451-463.
[16] Oshima G, Shinabukuro H, Nagasawa K. Peptide inhibitors of angiotensin-converting enzyme in digests of gelatin by bacterial collagenase [J]. Biochim Biophys Acta 1979, 566 (1): 128-137.
[17]吴建平,丁霄霖.食品蛋白质降血压肽的研究进展[J].中国粮油学报,1998,13(5):10-14.
[18] Maruyama S, Miyoshi S, Tanaka H. Angiotensin Iconverting enzyme inhibitor derived from Ficus carica[J]. Agric Biol Chem 1989, 53: 2763-2769.
[19] Kohama Y., Oka H., Yamamoto K., et al. Induction of angiotensin-converting enzyme inhibitory activity by acid-limited proteolysis of glyceraldehyde 3-phosphate dehydrogenase[J].Biochem Biophys Tes Commun.1989,161,456-460.
[20] Saito Y, Wanezaki K, Kawato A, et al. Structure and activity of angiotensin I- converting enzyme inhibitory peptides from sake and sake lees[J]. Biosci Biotech Biochem 1994, 58: 1767-1771.
[21] Okamoto A, Hanagata H, Kawamura Y, et al. Anti-hypertensive substances in fermented soybean, natto[J]. Plant Foods for Human Nutrition,1995,47(1): 39-47.
[22] Astawan M, Wahyuni M, Yasuhara T, Yamada K, Tadokoro T, Maekawa A. Effects of Angiotensin I-Converting Enzyme Inhibitory Substances Derived from Indonesian Dried-salted fish on Blood Pressure of Rats. Biosci Biotech Biochem 1995,59(3), 425-429.
[23] Rhyu MR, Nam YJ, Lee HY.Screening of angiotensin converting enzyme inhibitors in cereals and legumes. Foods Biotechnol 1996, 5: 334-337.
[24] Maeno M, Yamanoto N, Takano T. Identification of an Antihypertensive Peptide from Casein Hydrolysate Produced by a Proteinase from Lactobacillus helveticus CP790[J].Journal of Dairy Science,1996,79(8):1316-1321.
[25] Suetsuna K. Isolation and characterization of angiotensin I-converting enzyme inhibitor dipeptides derived from Allium sativum L (garlic)[J]. Journal of Nutritional Biochenmistry, 1998,9(7): 415-419.
[26] Arihara K. Peptide inhibitors for angiotensin I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle proteins.Meat Sci 2001,57:319-324.
[27] Wu J, Ding X. Hypotensive and Physiological Effect of Angiotensin Converting Enzyme Inhibitory Peptides Derived from Soy Protein on Spontaneously Hypertensive Rats[J]. Agric. Food Chem, 2001, 49 (1): 501-506.
[28] Marczak ED, Usui A, Fujiata H , et al. New Antihypertensive Peptides Isolated from Rapeseed. Peptides. 2003,24:791-798.
[29] Kuba M, Tawata S, et al. Angiotensin I-Converting Enzyme Inhibitory Peptides Isolated from Tofuyo Fermented Soybean Food[J]. Bioscience, Biotechnology, and Biochemistry. 2003, 67(6): 1278-1283.
[30] Hyoung Lee D, Ho Kim J, Sik Park J,et al. Isolation and characterization of a novel angiotensin I-converting enzyme inhibitory peptide derived from the edible mushroom Tricholoma giganteur[J]. Peptides,2004,25(4):621-627.
[31] He HL, Chen XL, Sun CY, Zhang YZ, Gao PJ. Preparation and functional evaluation of oligopeptide-enriched hydrolysate from shrimp (Acetes chinensis) treated with crude protease from Bacillus sp. SM98011. Bioresource Technology 2006,97:385-390.
[32] Yu,Y, Hu,J, Miyaguchi,Y, Bai,X, Du,Y, Lin,B. Isolation and characterization of angiotensin I-converting enzyme inhibitory peptides derived from porcine hemoglobin.Peptides 27,2950-2956.
[33]黄艳春,刘张虎,熊善柏.酶解鲢肉制取ACE抑制肽工艺条件的优化.食品科学, 2007,28 (2):181-184.
[34] Otte J, Shalaby M.A.S, Zakora M, Nielsen M.S. Fractionation and identification of ACE-inhibitory peptides fromα-lactalbumin andβ-casein produced by thermolysin-catalysed hydrolysis.[J] Int.Dairy.17,1460-1472.
[35] Lieselot Vercruysse, Guy Smagghe, Tanja Beckers, John Van Camp. Purification and identification of an angiotensin I converting enzyme (ACE) inhibitory peptide from the gastrointestinal hydrolysate of the cotton leafworm, Spodoptera littoralis. Process Biochemistry, 2008, 43(8): 900-904.
[36] Nakamura Y, Yamamoto N, Sakai K, et al. Purification and characterization of angiotensin I - converting enzyme inhibitory from sour milk. Journal of Dairy Science,1995,78:777-783.
[37] Nakamura Y, Yamamoto N, Sakai K, et al. Antihypertensive effect of sour milk and peptides isolated from it that are inhibitors to angiotensin I-converting enzyme. Journal of Dairy Science,1995,78:1253-1257.
[38] Nakamura Y, Masuda O, Takano T. Decrease of tissue angiotensin-Ⅰ-converting enzyme activity upon feeding sourmilk in spontaneously hypertensive rats[J]. Biosci Biotechnol Biochem,1996, 60(3): 488-489.
[39] TIINA JAUHIAINEN, RIITTA KORPELA, ANNIKA MAYRA MAKINEN. Information About the Evolus fermented milk[P]. Valio LTD.
[40] Maruyama S,Suzuki H. A peptide inhibitor ofangiotensinⅠ-converting enzyme in the tryptic hydrolysate of casein[J].Agric.Biol.Chem.,1982,46(5):1393-1394.
[41] Maruyama S,Nakagomi K,Tomizuka N, et al. AngiotenismⅠ-converting enzyme inhibitors derived from an enzymatic hydrolysate of casein.Ⅱ.Isolation of bradykinin-potentiating activity on the uterus and ileum of rats [J].Agric.Biol.Chem.,1985,49(5):1405-1409.
[42] Maruyama S,Mitachi H,Awaya J et al. AngiotenismⅠ-converting enzyme inhibitory activity of the C-terminal hexapeptide ofαs1-casein [J]. Agric.Biol.Chem.,1987,51(9):2557-2561
[43] Yamamoto N, Akino A, Takano T. Purification and specificity of a cell - wall associated proteinase from Lactobacillus helveticus CP790. Journal of Biochemistry,1993,114:740-745.
[44] Yamamoto N, MaenoM, Takano T. Purification and characterization of an antihypertensive peptide from a yogurt-like product fermented by Lactobacillus helveticus CPN4[J]. Journal of Dairy Science,1999,82:1388-1393.
[45] Kawase M,Hashimoto H,Hosoda M,et al.Effect of administration of fermented milk containing whey protein concentrate to rats and healthy men on serum lipids and blood pressure[J].J Dairy Sci,2000,83:255-263.
[46] Gobbetti M,Ferranti P,Smacchi E, et al..Production of angiotensin converting enzyme inhibitory peptides in fermented milks started by lactobacillus delbrueckii subsp.bulgaricus SS1 and lactococcus lactis subsp.cremoris FT4[J].Appl Environ Microbiol,2000,66:3898-3904.
[47] Saito T, Nakamura T, KitaZawa H, et al. Isolation and Structural Analysis of AntihyPertensive PePtides That Exist Naturally in Gouda Cheese[J].J.Dairry Sci.2000,83:1434-1440.
[48] Flambard,Benedicte.Peptides with antihypertensive properties[P].U.S.20050142166,2005.
[49] Muguerza B,Ramos M,Sanchez E, et al. Antihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk[J].International Dairy Journal, 2006,16: 61-69.
[50] Bütikofer U, Meyer J, Sieber R, et al. Quantification of the angiotensin-converting enzyme- inhibiting tripeptides Val-Pro-Pro and Ile-Pro-Pro in hard, semi-hard and soft cheeses[J]. International Dairy Journal, 2007, 17:968-975.
[51] Quirós A,Ramosa M,Muguerza B,et al.Identification of novel antihypertensive peptides in milk fermented with Enterococcus faecalis[J].International Dairy Joumal,2007,17(1):33-41.
[52]张佳程,王海燕,褚庆环.发酵乳中ACE抑制剂的超滤分离[J].中国乳品工业,2004,32(1):32-34.
[53]潘道东.酸乳中抗高血压肽的分离及其特性研究[J].食品科学,2005,26(1):205-210.
[54]王立平.瑞士乳杆菌酪蛋白源活性肽制备及其生理功效研究[D].北京:北京林业大学,2008.
[55] Muguerza B, Ramos M, Sanchez E, et al. Anrihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk [J].International Dairy Journal, 2006,16: 61-69.
[56] Seppo L K O, Suomalainen T, Korpela R. The effect of a Lactobacillus helveticus LBK-16H fermented milk on hypertension-a pilot study on humans [J].Milchwissenschaft, 2002,57: 124-127
[57]商靓舰,陈卫,田丰伟,等.具有血管紧张素转换酶抑制活性的乳酸菌筛选及特性研究[J].中国乳品工业,2006,34(4):24-28
[58]黄文利,陈卫,田丰伟,等.干酪乳杆菌Lc-15发酵乳血管紧张素转换酶体外抑制活性的研究[J].食品研究与开发,2006,27(10);17-21.
[59] Cheung H S,Wang F L,Ondetti M A,et al.Binding of peptide substrates and inhibitors of the angiotensin-converting enzyme[J]. Journal of Biology Chemistry,1980, 255:401-407.
[60] Moskowitz D W. Is‘somatic’angiotensin-I-converting enzyme a mechanosensor?[J].Diabetes Technology Therety,2003,4:841-858.
[61] PEDERSEN C , JONSSON H , LINDBERGJ E , et al . Microbiological characterization of wet wheat distillers’grain , with focus on isolation of lactobacilli with potential as probiotics[J] . Appl Environ Microbiol , 2004 , 70 (3) :1522-1527.
[62] CASEY P G, CASEY G D , GARDINER G E , et al . Isolation and characterization of anti- Salmonella lactic acid bacteria from the porcine gast rointestinal tract [J]. Lett Appl Microbiol ,2004 , 39 :431-438.
[63]吕兵,张国农,杨瑞欢.嗜酸乳杆菌生物学特性及其发酵乳的研究[J].中国乳品工业, 2002, 30 (5):37-39.
[64]张扬,袁杰利.模拟消化环境对益生菌制剂的影响[J].中国微生态学杂志,2003,15(5):253-255.
[65]中华人民共和国卫生部药典委员会.中华人民共和国药典[M].广州:广东科技出版社,1995:附录.54-65.
[66]张和平,张列兵.现代乳品工业手册[M].北京:中国轻工业出版社2005. 217.
[67]吕加平,董晓波,肖锐.发酵乳品中乳酸菌生物量测定方法的研究[J].肉品卫生1997,(2):3-7.
[68]赵新淮,冯志彪.蛋白质水解物水解度的测定[J].食品科学,1994,11:65-67.
[69] Church F C S H E, Proter D H,et al. Spectrophotometric assay using o-phthalaldyhyde for determination of proteolysis in milk and isolated milk protein [J]. Dairy Sci, 1983, 66:1219-1227.
[70]凌代文,东秀珠.乳酸细菌分类鉴定及试验方法[M].北京:中国轻工业出版社, 1999.
[71]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].科学出版社, 2001.
[72] R.E.布坎南, N.E.吉本斯,等.伯杰细菌鉴定手册(第八版)[M].北京:科学出版社, 1984.
[73]潘道东.酸乳中抗高血压肽的分离及其特性研究[J].食品科学,2005,26(1):205-209.
[74]潘道东.抗ACE活性之瑞士乳杆菌的筛选[J].食品科学,2007,28(2):145-148.
[75] Cushman D W C H S. Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung [J]. Biochem Pharmacol, 1971,20:1637-1648.
[76] Parrot S, Degraewe P, Cuna C, et al. In vitro study on digestion of peptides in Emmental cheese Analytical evaluation and influence on angiotensin I converting enzyme inhibitory peptides Nahrung/Food, 2003, 47 :87-94.
[77] Vermeirssen VV, camp JV, Decroos h, et al. The impact of fermentation and in vitro digestion on the formation of angiotensin-I-converting enzyme inhibitory activity from pea and whey protein [J]. Dairy Sci,2003, 86: 429-438.
[78] Hernandez-Ledesma B, Amigo L, Ramos M, et al Release of angiotensin converting enzyme-inhibitory peptides by simulated gastrointestinal digestion of infant formulas Int Dairy J, 2004,14 889-898.
[79]黎观红.食物蛋白源血管紧张素转化酶抑制肽的研究[D].博士学位论文.江南大学,2005.
[80] Virtanen T, Pihlanto A, Akkanen S, et al.Development of antioxidant activity in milk whey during fermentation with lactic acid bacteria[J]. Applied Microbiology,2007,102:106-115.
[81] Oomen AG, Hack A, Minkus M, et al. Comparison of fine in vitro digestion models to study the bioaccessibility of soil contaminants Emviron Sci Technol,2002, 36 3326-3334
[82]FitzGerald R J,Murray B A, Walsh G J.Hypotensive peptides from milk proteins[J].Journal of Nutrition,2004,134:980s-988s.
[83] Prior R L,Cao G.H.In vivo tatal antioxidant capacity:comparison of different analytical methods[J]. Free Radical Biology&Medicine,1999,27:1173-1181.
[84] Takano T.Milk derived peptides and hypertension reduction[J].International Dairy Journal,1998, 8:375-381.
[2] Julian Tudor Hart,Tom Fahey,Wendy Savage.高血压权威指南[M].石油工业出版社,2001,7.
[3]《中国高血压防治指南2005年修订版》,中国高血压防治指南起草委员会.
[4]郭胜才,冯友根.抗高血压药物研究新进展.中国药学杂志,2001, 36:583-587.
[5]徐贵成,霍保民,王秋风,等.通心络胶囊治疗风湿性心瓣膜病心力衰竭临床观察[J].浙江中西医结合志,2002,12(1):5-7.
[6]安茂竹,孙静,高景芳,等.血管紧张素转换酶抑制剂的不良反应及防治[J].中国新药杂志,1999, 8(5):403-404.
[7]吴建平.大豆降压肽的研制[D].博士学位论文.无锡:江南大学,1998.
[8]靳姝杰,袭荣刚,王晓波.血管紧张素转换酶抑制剂的不良反应和禁忌症[J].药学实践杂志,2000, 18(3):166-168.
[9]赵海珍,陆兆新,刘战民.天然食品来源的血管紧张素转换酶抑制肽的研究进展[J].中国生化药物杂志,2004,25(5):315-317.
[10]辛志宏,马海乐.食品蛋白质中降血压肽的功能与应用[J].食品与发酵工业,2003,29(8),32-36.
[11] Johnston JI,Franz VL.Renin-angiotensin system: a dual tissue and hormonal system for cardiovascular control[J]. J Hypertens,1992,10:S13-S26.
[12] Ferreira SH. Abradykinin-potentiating factor(BPF) present in the venom of Bothrops jararaca. Brit J Pharmacol,1965,24:163-169.
[13] Bakhle Y S.Conversion of angiotensin-Ⅰto angiotensin-Ⅱby cell free extracts of dog lung[J]. Nature 1968, 220 (5170):919-921.
[14] Ondetti M A, Williams N J, SaboEF, et al. Angiotensin-converting enzyme inhibitors from the Venom of Bothrops Jararaca.Isolation, elucidation of structure, and synthesis.[J] Biochemisty 1971,10(22):4033-4039.
[15] Cheung H S, Cushman D W. Inhibition of homogeneous angiotensin-converting enzyme of rabbit lung by synthetic venom peptides of Bothrops Jararaca[J]. Biochim Biophys Acta 1973, 293(2):451-463.
[16] Oshima G, Shinabukuro H, Nagasawa K. Peptide inhibitors of angiotensin-converting enzyme in digests of gelatin by bacterial collagenase [J]. Biochim Biophys Acta 1979, 566 (1): 128-137.
[17]吴建平,丁霄霖.食品蛋白质降血压肽的研究进展[J].中国粮油学报,1998,13(5):10-14.
[18] Maruyama S, Miyoshi S, Tanaka H. Angiotensin Iconverting enzyme inhibitor derived from Ficus carica[J]. Agric Biol Chem 1989, 53: 2763-2769.
[19] Kohama Y., Oka H., Yamamoto K., et al. Induction of angiotensin-converting enzyme inhibitory activity by acid-limited proteolysis of glyceraldehyde 3-phosphate dehydrogenase[J].Biochem Biophys Tes Commun.1989,161,456-460.
[20] Saito Y, Wanezaki K, Kawato A, et al. Structure and activity of angiotensin I- converting enzyme inhibitory peptides from sake and sake lees[J]. Biosci Biotech Biochem 1994, 58: 1767-1771.
[21] Okamoto A, Hanagata H, Kawamura Y, et al. Anti-hypertensive substances in fermented soybean, natto[J]. Plant Foods for Human Nutrition,1995,47(1): 39-47.
[22] Astawan M, Wahyuni M, Yasuhara T, Yamada K, Tadokoro T, Maekawa A. Effects of Angiotensin I-Converting Enzyme Inhibitory Substances Derived from Indonesian Dried-salted fish on Blood Pressure of Rats. Biosci Biotech Biochem 1995,59(3), 425-429.
[23] Rhyu MR, Nam YJ, Lee HY.Screening of angiotensin converting enzyme inhibitors in cereals and legumes. Foods Biotechnol 1996, 5: 334-337.
[24] Maeno M, Yamanoto N, Takano T. Identification of an Antihypertensive Peptide from Casein Hydrolysate Produced by a Proteinase from Lactobacillus helveticus CP790[J].Journal of Dairy Science,1996,79(8):1316-1321.
[25] Suetsuna K. Isolation and characterization of angiotensin I-converting enzyme inhibitor dipeptides derived from Allium sativum L (garlic)[J]. Journal of Nutritional Biochenmistry, 1998,9(7): 415-419.
[26] Arihara K. Peptide inhibitors for angiotensin I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle proteins.Meat Sci 2001,57:319-324.
[27] Wu J, Ding X. Hypotensive and Physiological Effect of Angiotensin Converting Enzyme Inhibitory Peptides Derived from Soy Protein on Spontaneously Hypertensive Rats[J]. Agric. Food Chem, 2001, 49 (1): 501-506.
[28] Marczak ED, Usui A, Fujiata H , et al. New Antihypertensive Peptides Isolated from Rapeseed. Peptides. 2003,24:791-798.
[29] Kuba M, Tawata S, et al. Angiotensin I-Converting Enzyme Inhibitory Peptides Isolated from Tofuyo Fermented Soybean Food[J]. Bioscience, Biotechnology, and Biochemistry. 2003, 67(6): 1278-1283.
[30] Hyoung Lee D, Ho Kim J, Sik Park J,et al. Isolation and characterization of a novel angiotensin I-converting enzyme inhibitory peptide derived from the edible mushroom Tricholoma giganteur[J]. Peptides,2004,25(4):621-627.
[31] He HL, Chen XL, Sun CY, Zhang YZ, Gao PJ. Preparation and functional evaluation of oligopeptide-enriched hydrolysate from shrimp (Acetes chinensis) treated with crude protease from Bacillus sp. SM98011. Bioresource Technology 2006,97:385-390.
[32] Yu,Y, Hu,J, Miyaguchi,Y, Bai,X, Du,Y, Lin,B. Isolation and characterization of angiotensin I-converting enzyme inhibitory peptides derived from porcine hemoglobin.Peptides 27,2950-2956.
[33]黄艳春,刘张虎,熊善柏.酶解鲢肉制取ACE抑制肽工艺条件的优化.食品科学, 2007,28 (2):181-184.
[34] Otte J, Shalaby M.A.S, Zakora M, Nielsen M.S. Fractionation and identification of ACE-inhibitory peptides fromα-lactalbumin andβ-casein produced by thermolysin-catalysed hydrolysis.[J] Int.Dairy.17,1460-1472.
[35] Lieselot Vercruysse, Guy Smagghe, Tanja Beckers, John Van Camp. Purification and identification of an angiotensin I converting enzyme (ACE) inhibitory peptide from the gastrointestinal hydrolysate of the cotton leafworm, Spodoptera littoralis. Process Biochemistry, 2008, 43(8): 900-904.
[36] Nakamura Y, Yamamoto N, Sakai K, et al. Purification and characterization of angiotensin I - converting enzyme inhibitory from sour milk. Journal of Dairy Science,1995,78:777-783.
[37] Nakamura Y, Yamamoto N, Sakai K, et al. Antihypertensive effect of sour milk and peptides isolated from it that are inhibitors to angiotensin I-converting enzyme. Journal of Dairy Science,1995,78:1253-1257.
[38] Nakamura Y, Masuda O, Takano T. Decrease of tissue angiotensin-Ⅰ-converting enzyme activity upon feeding sourmilk in spontaneously hypertensive rats[J]. Biosci Biotechnol Biochem,1996, 60(3): 488-489.
[39] TIINA JAUHIAINEN, RIITTA KORPELA, ANNIKA MAYRA MAKINEN. Information About the Evolus fermented milk[P]. Valio LTD.
[40] Maruyama S,Suzuki H. A peptide inhibitor ofangiotensinⅠ-converting enzyme in the tryptic hydrolysate of casein[J].Agric.Biol.Chem.,1982,46(5):1393-1394.
[41] Maruyama S,Nakagomi K,Tomizuka N, et al. AngiotenismⅠ-converting enzyme inhibitors derived from an enzymatic hydrolysate of casein.Ⅱ.Isolation of bradykinin-potentiating activity on the uterus and ileum of rats [J].Agric.Biol.Chem.,1985,49(5):1405-1409.
[42] Maruyama S,Mitachi H,Awaya J et al. AngiotenismⅠ-converting enzyme inhibitory activity of the C-terminal hexapeptide ofαs1-casein [J]. Agric.Biol.Chem.,1987,51(9):2557-2561
[43] Yamamoto N, Akino A, Takano T. Purification and specificity of a cell - wall associated proteinase from Lactobacillus helveticus CP790. Journal of Biochemistry,1993,114:740-745.
[44] Yamamoto N, MaenoM, Takano T. Purification and characterization of an antihypertensive peptide from a yogurt-like product fermented by Lactobacillus helveticus CPN4[J]. Journal of Dairy Science,1999,82:1388-1393.
[45] Kawase M,Hashimoto H,Hosoda M,et al.Effect of administration of fermented milk containing whey protein concentrate to rats and healthy men on serum lipids and blood pressure[J].J Dairy Sci,2000,83:255-263.
[46] Gobbetti M,Ferranti P,Smacchi E, et al..Production of angiotensin converting enzyme inhibitory peptides in fermented milks started by lactobacillus delbrueckii subsp.bulgaricus SS1 and lactococcus lactis subsp.cremoris FT4[J].Appl Environ Microbiol,2000,66:3898-3904.
[47] Saito T, Nakamura T, KitaZawa H, et al. Isolation and Structural Analysis of AntihyPertensive PePtides That Exist Naturally in Gouda Cheese[J].J.Dairry Sci.2000,83:1434-1440.
[48] Flambard,Benedicte.Peptides with antihypertensive properties[P].U.S.20050142166,2005.
[49] Muguerza B,Ramos M,Sanchez E, et al. Antihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk[J].International Dairy Journal, 2006,16: 61-69.
[50] Bütikofer U, Meyer J, Sieber R, et al. Quantification of the angiotensin-converting enzyme- inhibiting tripeptides Val-Pro-Pro and Ile-Pro-Pro in hard, semi-hard and soft cheeses[J]. International Dairy Journal, 2007, 17:968-975.
[51] Quirós A,Ramosa M,Muguerza B,et al.Identification of novel antihypertensive peptides in milk fermented with Enterococcus faecalis[J].International Dairy Joumal,2007,17(1):33-41.
[52]张佳程,王海燕,褚庆环.发酵乳中ACE抑制剂的超滤分离[J].中国乳品工业,2004,32(1):32-34.
[53]潘道东.酸乳中抗高血压肽的分离及其特性研究[J].食品科学,2005,26(1):205-210.
[54]王立平.瑞士乳杆菌酪蛋白源活性肽制备及其生理功效研究[D].北京:北京林业大学,2008.
[55] Muguerza B, Ramos M, Sanchez E, et al. Anrihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk [J].International Dairy Journal, 2006,16: 61-69.
[56] Seppo L K O, Suomalainen T, Korpela R. The effect of a Lactobacillus helveticus LBK-16H fermented milk on hypertension-a pilot study on humans [J].Milchwissenschaft, 2002,57: 124-127
[57]商靓舰,陈卫,田丰伟,等.具有血管紧张素转换酶抑制活性的乳酸菌筛选及特性研究[J].中国乳品工业,2006,34(4):24-28
[58]黄文利,陈卫,田丰伟,等.干酪乳杆菌Lc-15发酵乳血管紧张素转换酶体外抑制活性的研究[J].食品研究与开发,2006,27(10);17-21.
[59] Cheung H S,Wang F L,Ondetti M A,et al.Binding of peptide substrates and inhibitors of the angiotensin-converting enzyme[J]. Journal of Biology Chemistry,1980, 255:401-407.
[60] Moskowitz D W. Is‘somatic’angiotensin-I-converting enzyme a mechanosensor?[J].Diabetes Technology Therety,2003,4:841-858.
[61] PEDERSEN C , JONSSON H , LINDBERGJ E , et al . Microbiological characterization of wet wheat distillers’grain , with focus on isolation of lactobacilli with potential as probiotics[J] . Appl Environ Microbiol , 2004 , 70 (3) :1522-1527.
[62] CASEY P G, CASEY G D , GARDINER G E , et al . Isolation and characterization of anti- Salmonella lactic acid bacteria from the porcine gast rointestinal tract [J]. Lett Appl Microbiol ,2004 , 39 :431-438.
[63]吕兵,张国农,杨瑞欢.嗜酸乳杆菌生物学特性及其发酵乳的研究[J].中国乳品工业, 2002, 30 (5):37-39.
[64]张扬,袁杰利.模拟消化环境对益生菌制剂的影响[J].中国微生态学杂志,2003,15(5):253-255.
[65]中华人民共和国卫生部药典委员会.中华人民共和国药典[M].广州:广东科技出版社,1995:附录.54-65.
[66]张和平,张列兵.现代乳品工业手册[M].北京:中国轻工业出版社2005. 217.
[67]吕加平,董晓波,肖锐.发酵乳品中乳酸菌生物量测定方法的研究[J].肉品卫生1997,(2):3-7.
[68]赵新淮,冯志彪.蛋白质水解物水解度的测定[J].食品科学,1994,11:65-67.
[69] Church F C S H E, Proter D H,et al. Spectrophotometric assay using o-phthalaldyhyde for determination of proteolysis in milk and isolated milk protein [J]. Dairy Sci, 1983, 66:1219-1227.
[70]凌代文,东秀珠.乳酸细菌分类鉴定及试验方法[M].北京:中国轻工业出版社, 1999.
[71]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].科学出版社, 2001.
[72] R.E.布坎南, N.E.吉本斯,等.伯杰细菌鉴定手册(第八版)[M].北京:科学出版社, 1984.
[73]潘道东.酸乳中抗高血压肽的分离及其特性研究[J].食品科学,2005,26(1):205-209.
[74]潘道东.抗ACE活性之瑞士乳杆菌的筛选[J].食品科学,2007,28(2):145-148.
[75] Cushman D W C H S. Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung [J]. Biochem Pharmacol, 1971,20:1637-1648.
[76] Parrot S, Degraewe P, Cuna C, et al. In vitro study on digestion of peptides in Emmental cheese Analytical evaluation and influence on angiotensin I converting enzyme inhibitory peptides Nahrung/Food, 2003, 47 :87-94.
[77] Vermeirssen VV, camp JV, Decroos h, et al. The impact of fermentation and in vitro digestion on the formation of angiotensin-I-converting enzyme inhibitory activity from pea and whey protein [J]. Dairy Sci,2003, 86: 429-438.
[78] Hernandez-Ledesma B, Amigo L, Ramos M, et al Release of angiotensin converting enzyme-inhibitory peptides by simulated gastrointestinal digestion of infant formulas Int Dairy J, 2004,14 889-898.
[79]黎观红.食物蛋白源血管紧张素转化酶抑制肽的研究[D].博士学位论文.江南大学,2005.
[80] Virtanen T, Pihlanto A, Akkanen S, et al.Development of antioxidant activity in milk whey during fermentation with lactic acid bacteria[J]. Applied Microbiology,2007,102:106-115.
[81] Oomen AG, Hack A, Minkus M, et al. Comparison of fine in vitro digestion models to study the bioaccessibility of soil contaminants Emviron Sci Technol,2002, 36 3326-3334
[82]FitzGerald R J,Murray B A, Walsh G J.Hypotensive peptides from milk proteins[J].Journal of Nutrition,2004,134:980s-988s.
[83] Prior R L,Cao G.H.In vivo tatal antioxidant capacity:comparison of different analytical methods[J]. Free Radical Biology&Medicine,1999,27:1173-1181.
[84] Takano T.Milk derived peptides and hypertension reduction[J].International Dairy Journal,1998, 8:375-381.