玉米蛋白水解物体外消化产物的抗氧化及ACE抑制活性研究
|
摘要
国内外研究表明,玉米醇溶蛋白水解物具有良好的体外抗氧化能力以及ACE抑制活性,但是有关玉米醇溶蛋白水解物体内抗氧化及ACE抑制活性的研究较少。本论文主要探讨玉米醇溶蛋白水解物抗氧化及ACE抑制活性的耐消化性,以及抗氧化能力与ACE抑制活性间的相关性。本论文的研究对于促进玉米蛋白在食品加工领域及保健食品领域的应用具有重要的理论和应用价值。
本论文首先研究了玉米醇溶蛋白的碱性蛋白酶解产物(ZH)的抗氧化性,利用体外两阶段消化模型(在37 oC,胃蛋白酶作用1 h后由胰蛋白酶继续消化2 h)研究ZH的体内抗氧化特性。以ABTS+·自由基清除能力,DPPH·自由基清除能力,还原能力和Cu2+螯合能力为抗氧化指标,并利用氨基酸自动分析仪和高效凝胶排阻色谱(HPSEC)来测定消化过程中游离氨基酸和分子量分布的变化。结果表明,ZH具有很强的ABTS+·自由基清除能力、Cu2+螯合能力以及还原能力。在5 mg/mL时,ZH的还原能力是0.01 mg/mL Vc的2倍左右,ABTS+·自由基清除能力是0.01 mg/mL Vc的7倍左右,其Cu2+螯合能力比0.1 mg/mL BHA的螯合能力高5倍多(P < 0.05)。经过体外消化,ZH的还原能力几乎提高了一倍,并且保留了原有的ABTS+·自由基清除能力以及Cu2+螯合能力(P < 0.05)。低浓度的ZH消化产物(1 mg/mL)对ABTS+·的清除能力比8 mg/mL的消化产物对DPPH·自由基的清除能力高出近50倍的现象表明,ABTS+·自由基清除能力的测定更适用于水溶性肽抗氧化的评价。
论文利用紫外分光光度法测定ZH及其体外消化产物的ACE抑制活性,通过SephadexG-15凝胶柱对ZH消化终产物进行分离,以ABTS+·自由基清除能力,还原能力,Cu2+螯合能力以及ACE抑制率为检测指标,研究了分离组分的抗氧化及ACE抑制活性,并探讨了氨基酸组成和分子量分布对于各组分抗氧化能力以及ACE抑制活性的影响。结果表明,ZH具有很好的ACE抑制活性(8 mg/mL蛋白浓度时抑制率达到80.6±2.67%)。经过体外消化,产物仍然具有良好的ACE抑制活性(83.8±2.03%)。ZH消化终产物经凝胶柱分离后得到四个组分,其中2号组分中分子量在120-568 Da的小肽占77.07%,具有抗氧化能力的氨基酸,碱性氨基酸,支链氨基酸以及疏水性氨基酸含量很高,具有最强的ABTS+·自由基清除能力,还原能力,金属离子螯合能力以及ACE抑制活性。
Zein protein hydrolysate showed a strong antioxidant activity and ACE-inhibitory activity, but little is known about the in vivo antioxidant activity and ACE-inhibitory activity. The objective of this study was to assess the survivability of antioxidative activity and ACE-inhibitory activity of Alcalase-treated zein hydrolysate (ZH), and the relationship between antioxidant activity and ACE-inhibitory activity.
Firstly, the antioxidative activity of zein hydrolysate treated by Alcalase (ZH) and the antioxidant potential of ZH during a two-stage (1 h pepsin→2 h pancreatin, 37 oC) in vitro digestion was studied. Free Amino acid composition, 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+·) and 1,1-diphenyl-2-picrylhydrazyl (DPPH?) free radical scavenging activity, reducing power, and Cu2+ chelation ability were tested to determine the antioxidant efficacy of ZH. High-performance size exclusion chromatography (HPSEC) was run to evaluate the molecular weight of ZH digests. Results showed that ZH exhibited a strong ABTS+·free radical scavenging activity, reducing power, and Cu2+ chelation ability. The antioxidant activity of ZH (5 mg/mL) was exceeded (P < 0.05) that of 0.01 mg/mL of ascorbic acid and 0.1 mg/mL BHA. After in vitro digestion, the reducing power of ZH increased one-fold (P < 0.05), the ABTS+·free radical scavenging activity and Cu2+ chelation ability were fully recovered when compared with nondigested ZH. The ZH samples tested in the ABTS+·system, though at a lower concentration (1 mg/mL), had a Trolox equivalence of approximately 50 times that of ZH sample (8 mg/mL) tested in the DPPH? system, suggested that the ABTS assay was an appropriate method for the measurement of antioxidant activity of water-soluble proteins and peptides in an aqueous solution.
The ACE-inhibitory activity of ZH and it’s digests was determined by Cushman Ultraviolet spectrophotometer. Sephadex G-15 gel filtration was used to separate the final digest of ZH into fractions. ABTS+·free radical scavenging activity, reducing power, Cu2+ chelation ability and ACE-inhibitory activity were tested to determine the antioxidant and ACE-inhibitory efficacy of each fractions, the effect of Amino acid composition and the molecular weight profile was also considered. Results showed that ZH can inhibit ACE activity by 80.6±2.67% at 8 mg/mL, and also exhibited a good ACE-inhibitory activity after in vitro digestion. Four fractions were yielded after Sephadex G-15 gel filtration, of which peptides with MW120-568 Da in fraction 2 representing 70.07% of the total mass, the composition of basic amino acid, branched-chain amino acid, hydrophobic amino acid and the amino acid with antioxidant activity was higher in fraction 2, showed the strongest antioxidant activity and ACE-inhibitory activity.
本论文首先研究了玉米醇溶蛋白的碱性蛋白酶解产物(ZH)的抗氧化性,利用体外两阶段消化模型(在37 oC,胃蛋白酶作用1 h后由胰蛋白酶继续消化2 h)研究ZH的体内抗氧化特性。以ABTS+·自由基清除能力,DPPH·自由基清除能力,还原能力和Cu2+螯合能力为抗氧化指标,并利用氨基酸自动分析仪和高效凝胶排阻色谱(HPSEC)来测定消化过程中游离氨基酸和分子量分布的变化。结果表明,ZH具有很强的ABTS+·自由基清除能力、Cu2+螯合能力以及还原能力。在5 mg/mL时,ZH的还原能力是0.01 mg/mL Vc的2倍左右,ABTS+·自由基清除能力是0.01 mg/mL Vc的7倍左右,其Cu2+螯合能力比0.1 mg/mL BHA的螯合能力高5倍多(P < 0.05)。经过体外消化,ZH的还原能力几乎提高了一倍,并且保留了原有的ABTS+·自由基清除能力以及Cu2+螯合能力(P < 0.05)。低浓度的ZH消化产物(1 mg/mL)对ABTS+·的清除能力比8 mg/mL的消化产物对DPPH·自由基的清除能力高出近50倍的现象表明,ABTS+·自由基清除能力的测定更适用于水溶性肽抗氧化的评价。
论文利用紫外分光光度法测定ZH及其体外消化产物的ACE抑制活性,通过SephadexG-15凝胶柱对ZH消化终产物进行分离,以ABTS+·自由基清除能力,还原能力,Cu2+螯合能力以及ACE抑制率为检测指标,研究了分离组分的抗氧化及ACE抑制活性,并探讨了氨基酸组成和分子量分布对于各组分抗氧化能力以及ACE抑制活性的影响。结果表明,ZH具有很好的ACE抑制活性(8 mg/mL蛋白浓度时抑制率达到80.6±2.67%)。经过体外消化,产物仍然具有良好的ACE抑制活性(83.8±2.03%)。ZH消化终产物经凝胶柱分离后得到四个组分,其中2号组分中分子量在120-568 Da的小肽占77.07%,具有抗氧化能力的氨基酸,碱性氨基酸,支链氨基酸以及疏水性氨基酸含量很高,具有最强的ABTS+·自由基清除能力,还原能力,金属离子螯合能力以及ACE抑制活性。
Zein protein hydrolysate showed a strong antioxidant activity and ACE-inhibitory activity, but little is known about the in vivo antioxidant activity and ACE-inhibitory activity. The objective of this study was to assess the survivability of antioxidative activity and ACE-inhibitory activity of Alcalase-treated zein hydrolysate (ZH), and the relationship between antioxidant activity and ACE-inhibitory activity.
Firstly, the antioxidative activity of zein hydrolysate treated by Alcalase (ZH) and the antioxidant potential of ZH during a two-stage (1 h pepsin→2 h pancreatin, 37 oC) in vitro digestion was studied. Free Amino acid composition, 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+·) and 1,1-diphenyl-2-picrylhydrazyl (DPPH?) free radical scavenging activity, reducing power, and Cu2+ chelation ability were tested to determine the antioxidant efficacy of ZH. High-performance size exclusion chromatography (HPSEC) was run to evaluate the molecular weight of ZH digests. Results showed that ZH exhibited a strong ABTS+·free radical scavenging activity, reducing power, and Cu2+ chelation ability. The antioxidant activity of ZH (5 mg/mL) was exceeded (P < 0.05) that of 0.01 mg/mL of ascorbic acid and 0.1 mg/mL BHA. After in vitro digestion, the reducing power of ZH increased one-fold (P < 0.05), the ABTS+·free radical scavenging activity and Cu2+ chelation ability were fully recovered when compared with nondigested ZH. The ZH samples tested in the ABTS+·system, though at a lower concentration (1 mg/mL), had a Trolox equivalence of approximately 50 times that of ZH sample (8 mg/mL) tested in the DPPH? system, suggested that the ABTS assay was an appropriate method for the measurement of antioxidant activity of water-soluble proteins and peptides in an aqueous solution.
The ACE-inhibitory activity of ZH and it’s digests was determined by Cushman Ultraviolet spectrophotometer. Sephadex G-15 gel filtration was used to separate the final digest of ZH into fractions. ABTS+·free radical scavenging activity, reducing power, Cu2+ chelation ability and ACE-inhibitory activity were tested to determine the antioxidant and ACE-inhibitory efficacy of each fractions, the effect of Amino acid composition and the molecular weight profile was also considered. Results showed that ZH can inhibit ACE activity by 80.6±2.67% at 8 mg/mL, and also exhibited a good ACE-inhibitory activity after in vitro digestion. Four fractions were yielded after Sephadex G-15 gel filtration, of which peptides with MW120-568 Da in fraction 2 representing 70.07% of the total mass, the composition of basic amino acid, branched-chain amino acid, hydrophobic amino acid and the amino acid with antioxidant activity was higher in fraction 2, showed the strongest antioxidant activity and ACE-inhibitory activity.
引文
1. Xiong YL, Decker EA. Alterations in muscle protein functionality by oxidative and antioxidative processes. J. Muscle Foods, 1995(6):139-160
2. Chen H M, Muramoto K, Yamauchi F, et al. Antioxidative properties of histidine-containing peptides designed from peptide fragments found in the digests of a soybean protein. J. Agric. Food Chem, 1998(46):49-53
3. Pena-Ramos E A, Xiong Y L. Antioxidative activity of soy protein hydrolysates in a liposomal system. J. Food Sci, 2002(67):2952-2956
4. Kong B, Xiong Y L. Antioxidant activity of zein hydrolysates in a liposome system and the possible mode of action. J. Agric. Food Chem, 2006(54):6059-6068
5. Xu L, Zhang X Z, GUO Y, et al. Preparation and Antioxidative Effects of Corn Peptides. J. CHEM. RES. CHINESE U, 2002,18(3):299-302
6. Xuli, Li Xiang-Lu, Wu Xiao-Xia et al. Preparation and Structural Characterization of a New Corn Antioxidative Peptide. J. CHEM. RES. CHINESE U,2004,25(3):466-469
7. Wang L, Xiong Y L. Inhibition of lipid oxidation in cooked beef patties by hydrolyzed potato protein is related to its reducing and radical scavenging ability. J. Agric. Food Chem, 2005(53): 9186-9192
8. Pena-Ramos E A, Xiong Y L. Antioxidative activity of whey protein hydrolysates in a liposomal system. J. Dairy Sci, 2001(84):2577-2583
9. Pena-Ramos E A, Xiong Y L, Arteaga G E. Fractionation and characterization for antioxidant activity of hydrolyzed whey protein. J. Sci. Food Agric, 2004(84):1908-1918
10. Zhu K X, Zhou H M, Qian H F. Antioxidant and free radical-scavenging activities of wheat germ protein hydrolysates (WGPH) prepared with alcalase. J. Process Biochem,2006(41): 1296-1302
11. Wang J S, Zhao M M, Zhao Q Z, et al. Antioxidant properties of papain hydrolysates of wheat gluten in different oxidation systems. J. Food Chem,2007 (101):1658-1663
12. Davalos A, Miguel M, Bartolome B, et al. Antioxidant activity of peptides derived from egg white proteins by enzymatic hydrolysis. J. Food Prot,2004(67):1939-1944
13. Mendis, E; Rajapakse, N; Byun, H.-G; et al. Investigation of jumbo squid (Dosidicus gigas) skin gelatin peptides for their in vitro antioxidant effects. J. Life Sci, 2005(77):2166-2178
14. Rajapakse N,Mendis E,Byun H G,et al. Purification and in vitro antioxidative effects of giant squid muscle peptides on free radical-mediated oxidative systems .J. Nutr Biochem, 2005, 16(9):562-569
15.夏向东,吕飞杰,台建祥.抗氧化剂的功效及体外抗氧化活性的评价[J].食品科学,2001,22(1):89-93
16.郑晶泉.抗氧化剂抗氧化实验研究进展[J].国外医学卫生学分册,2000,27(1):37-40
17.徐清萍,钟桂芳,孟君.抗氧化剂抗氧化方法研究进展[J].食品工程,2007(2):23-25
18. Block G, Patterson B H, Subar A F. Fruit, vegetables, and cancer prevention: a review of the epidemiological evidence. J. Nutr. Cancer, 1992 (18):11-29
19. Michel Soulié. What is the Role of Surgery for Locally Advanced Disease. J. European Urology Supplements, 2008(7):400-405
20.童荣生.自由基与心血管疾病[J].现代临床医学,2007,33(2):188-191
21. Willcox J K, Ash S, Catignani G. Antioxidants and prevention of chronic disease. J. Food Sci. Nutr, 2004 (44): 275-295
22.玄国东.米糟蛋白提取及酶法制备抗氧化活性肽及降血压活性肽的研究[D]:[博士学位论文],浙江:浙江大学,2005
23.吴亚梅,陈健,李维峰.玉米蛋白粉深加工应用的新进展[J].现代食品科技,2007,25(4): 97-100
24. Blanca Hernández-Ledesma,Lourdes Amigo, Isidra Recio, et al. ACE-inhibitory and radical-scavenging activity of peptides derived fromβ-lactoglobulin f(19-25). Interactions with ascorbic acid. J. Agric. Food Chem, 2007(55):3392-3397
25. Anne Pihlanto, Sari Akkanen, Hannu J. Korhonen. ACE-inhibitory and antioxidant properties of potato .J. Food Chemistry, 2008(109):104-112
26.张晓峰.腐乳发酵及其抗氧化和ACE抑制活性研究[D]:[博士学位论文].北京:中国农业大学,2006
27.郑喜群.玉米黄粉的酶解工艺与抗氧化活性肽的制备[D]:[博士学位论文].北京:中国农业大学, 2006
28. Shukla R, Cheryan M. Zein: the industrial protein from corn. J. Ind. Crops Prod,2001(13): 171–192
29.常亮,韩恩山,胡建修,等.玉米醇溶蛋白的研究及应用[J].化工中间体,2006(1):10-12
30.白艳菊,朱永军,代英杰,等.玉米醇溶蛋白的提取和应用[J].黑龙江农业科学,2001(1): 34-35
31陈石良,吴建平,谷文英.高F值寡肽的研究进展[J].食品与机械,1998(2):12-14
32.郭维静.玉米蛋白水解制取玉米肽的研究.[D]:[硕士学位论文],大连:大连理工大学
33.鲁晓翔,陈新华,唐津忠.酶法改性玉米蛋白功能特性的研究[J].食品科学,2000,21(12): 13-15
34.王遂,尤旭.微生物蛋白酶对玉米皮蛋白水解作用的研究[J].食品工业科技,2000,21(4): 22-24
35. Seiko Yano, Kazumasa Suzuki, Gunki Funatsu. lsolation from a-zein of themokysin peptides with Angiotensin1-converting Enzyme In hibitory Activity. J. Biosci. Biotech. Biochem,1996,60(4): 661-663
36.王梅,戴军.高效凝胶过滤色谱法测定玉米蛋白酶解物寡肽的分子量分布[J].食品与发酵工业,1998,24(5):25-27
37.王梅,谷文英.酶解玉米黄粉蛋白制备可溶性肽[J].粮油食品科技,1999,7(1):1-3
38.郑冬梅,孔保华,李升福.玉米蛋白及其水解肽的研究动态[J].食品与发酵工业,2002, 28(11):55-59
39.郑冬梅,李升福,孔保华.玉米蛋白水解条件的优化研究[J].食品科学,2002,23(3):52-56
40.赵金兰.用玉米渣生产降血压活性肽[J].食品与机械,1998(5):7-8
41.何慧,谢笔钧,杨卓.大豆蛋白和玉米蛋白酶解肽及其活性研究[J].粮油食品科技,2002,
10(1):14-16
42. Xu L, Zhang X Z, GUO Y, et al. A Study on Bioactivity of Corn Peptides with Low Molecular Weight Effect of an In take of them on Alcohol Metabolism in Rats. J. CHEM. RES. CH IN ESE U, 2003, 19(2): 180-182
43.李晶.玉米肽抗疲劳作用的实验研究[J].食品与机械, 2004, 20(1): 11-13
44.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(8):156-158
45.张强,阚国仕,陈红漫.玉米抗氧化肽的分离制备及其体外抗氧化活性的研究[J].中国粮油学报,2005,20(5):36-39
46.刘萍.酶解玉米蛋白提取ACE抑制肽的研究[D]:[硕士学位论文],江苏:江南大学,2006
47.吴炜亮,吴国杰,梁道双等. ACE抑制肽的生理功能和研究进展[J].现代食品技,2006, 22(3):251-254
48.罗郁中.台灣六種野生植物果實抗氧化功能評估[D]:[硕士学位论文],台湾:国立大,民国九十二年
49. [109]熊双丽,李安林,金征宇.猪喉软骨硫酸软骨素体外超氧阴离子自由基清除机理探讨[J].食品科学,2008,29(2):362-365
50.程云辉.麦胚蛋白酶解物的制备、结构及其生物活性功能的研究[D]:[博士学位论文].江苏:江南大学,2006
51.严群芳.大豆抗氧化肽的分离及其生物活性研究[D]:[硕士学位论文].江苏:南京农业大学,2006
52.张铁鹰,汪儆.单胃动物体外消化模拟技术研究进展[J].动物营养学报,2005,17(2):1-8
53.刘选珍,乐国伟,施用晖.饲料蛋白质体外消化率及其影响因素的研究[J].四川农业大学学报,1996(14):51-57
54. Arlene Wolzak, Ricardo Bressani, Roberto Gomez Brenes. A comparison of in vivo and in vitro estimates of protein digestibility of native and thermally processed vegetable proteins. J. Qual Plant Plant Foods ttum Nutr, 1981(31):31-43
55.王显生,杨晓泉,唐传核.大麻蛋白的营养评价[J].现代食品科技,2005,23(7):10-14
56.汤国营,戴军,蔡木易,等.大豆低聚肽分子质量分布的测定及体外消化实验[J].食品与发酵工业,2006,32(5):131-133
57. Cyrille Krul, Anja Luiten-Schuite, Aschwin Tenfelde, et al. Antimutagenic activity of green tea and black tea extracts studied in a dynamic in vitro gastrointestinal model. J. Mutation Research,2001(474):71-85
58. Blanca Hernández-Ledesma, Lourdes Amigo, Mercedes Ramos,et al. Release of angiotensin converting enzyme-inhibitory peptides by simulated gastrointestinal digestion of infant formulas. J. International Dairy Journal,2004(14):889-898
59. Blanca Hernández-Ledesma, Ana Quirós, Lourdes Amigo, et al. Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin. J. International Dairy Journal, 2007(17):42-49
60. Lo, W. M. Y; Li-Chan, E. C. Angiotensin I converting enzyme inhibitory peptides from in vitro pepsin-pancreatin digestion of soy protein. J. Agric. Food Chem, 2005(53):3369-3376
61. Marta Miguel, María J. Alonso, Mercedes Salaices, et al. Antihypertensive, ACE-inhibitory and vasodilator properties of an egg white hydrolysate: Effect of a simulated intestinal digestion. J. Food Chemistry, 2007 (104):163-168
62. Zhong-Ji Qian, Won-Kyo Jung, Hee-Guk Byun, et al. Protective effect of an antioxidative peptide purified from gastrointestinal digests of oyster, Crassostrea gigas against free radical induced DNA damage. J. Bioresource Technology, 2008(99):3365-3371
63.杨梅琳.蚕蛹蛋白的酶法水解及其产物的抗氧化性研究[D]:[硕士学位论文].江苏:江南大学,2006
64. Garrett, D. A; Failla, M. L; Sarama, R. J. Development of an in vitro digestive method to assess carotenoid bioavailability from meals. J. Agric. Food Chem, 1999(47):4301-4309
65. Adler-Nissen, J. Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. J. Agric. Food Chem, 1979(27):1256-1262
66. Spellman, D.; McEvoy, E.; O’Cuinn, G.; FitzGerald, R. J. Proteinase and exopeptidase hydrolysis of whey protein: Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis. J. Intl. Dairy, 2003(13):447-453
67.余冰宾,生物化学实验指导[M].第1版.北京:清华大学出版社,2004.133-136
68. Pellegrini, N. R; Proteggente, N; Pannala, A;et al. Antioxidant activity applying an improved ABTS radical cation decolorization assay. J. Free Radical Biol. Med, 1999(26):1231-1237
69. Wu, H.-C; Chen, H.-M; Shiau, C.-Y. Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus). J. Food Res. Intl, 2003(36): 949-957
70. Marcuse, R. Antioxidative effect of amino-acids. J. Nature, 1960(186):886-887
71. Saiga A, Tanabe S, Nishimura T. Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment. J. Agric. Food Chem, 2003(51):3661-3667
72. Decker, E. A; Crum, A. D; Calvert, J. T. Differences in the antioxidant mechanism of carnosine in the presence of copper and iron. J. Agric. Food Chem, 1992(40):756-759
73.布冠好,杨国宇,李宏基等.肌肽对猪肉匀浆的氧化抑制作用研究[J].河南工业大学学报(自然科学版),2006,27(3):62-65
74. Chen H M, Muramoto K, Yamauchi F. Structural analysis of antioxidative peptides from soybeanβ-conglycinin. J. Agric. Food Chem, 1995(43):574-578.
75.豆康宁.酶解大豆分离蛋白制取大豆肽及其抗氧化性研究[D]:[硕士学位论文].河南:河南工业大学,2007
76.姜瞻梅.酶解酪蛋白生产ACE抑制肽[D]:[硕士学位论文].哈尔滨:东北农业大学,2003
77.张宇昊.花生短肽制备及其功能活性研究[D]:[博士学位论文].北京:中国农业科学院. 2007
78.倪莉,陶冠军,戴军,等.血管紧张素转化酶活性抑制剂丝素肽的分离、纯化和结构鉴定[J].色谱,2001,19(3):222-225
79.于娅,杨瑞金,王璋.牡蛎功能短肽的制备及ACE抑制活性[J].无锡轻工大学学报,2004,23(2):49-52
80.管骁姚惠源.酶法制备燕麦麸蛋白ACE抑制肽的研究.食品与机械,2006, 22(6):12-15
81.刘志国,吴琼,吕玲肖,等.酶解米糠蛋白分离提取ACE抑制肽及其结构研究[J].食品科学, 2007,28(3):223-227
82.吴炜亮,吴国杰,梁道双等.ACE抑制肽的生理功能和研究进展[J].现代食品技,2006,22(3):251-254
83.任维栋.猪骨胶原蛋白酶解物中血管紧张素转换酶抑制剂的提纯[J].生物化学杂志,1996,l2(6):693-697
84.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(8):156-158
85.张莉莉,严群芳,王恬.大豆生物活性肽的分离及其抗氧化活性研究[J].食品科学,2007, 28(5):208-211
86.徐力,赵忠岩,李鸿梅等.大豆蛋白的小分子酶解产物抗氧化活性研究.吉林农业大学学报, 2007,29(1):48-52
87.郭红,李茂辉,阎萍,等.大豆小分子肽抗氧化效应的初步研究[J].中华医学研究杂志,2007,7(3):196-198
88. Chen H M, Muramoto K, Yamauchi F, et al. Antioxidant activity of designed peptides based on the antioxidative peptide isolated from digests of a soybean protein. J. Agric. Food Chem, 1996(44):2619-2623
89. JE J Y,PARK P J,KIM S K. Antioxidant activity of a peptide isolated from alaska pollack (Theragra chalcogramma) frame protein hydrolysate.J.Food Res Int, 2005,38(1):45-50
90.徐明生.鸡蛋卵白蛋白酶解物抗氧化肽研究[D]:[博士学位论文].陕西:陕西师范大学, 2006
2. Chen H M, Muramoto K, Yamauchi F, et al. Antioxidative properties of histidine-containing peptides designed from peptide fragments found in the digests of a soybean protein. J. Agric. Food Chem, 1998(46):49-53
3. Pena-Ramos E A, Xiong Y L. Antioxidative activity of soy protein hydrolysates in a liposomal system. J. Food Sci, 2002(67):2952-2956
4. Kong B, Xiong Y L. Antioxidant activity of zein hydrolysates in a liposome system and the possible mode of action. J. Agric. Food Chem, 2006(54):6059-6068
5. Xu L, Zhang X Z, GUO Y, et al. Preparation and Antioxidative Effects of Corn Peptides. J. CHEM. RES. CHINESE U, 2002,18(3):299-302
6. Xuli, Li Xiang-Lu, Wu Xiao-Xia et al. Preparation and Structural Characterization of a New Corn Antioxidative Peptide. J. CHEM. RES. CHINESE U,2004,25(3):466-469
7. Wang L, Xiong Y L. Inhibition of lipid oxidation in cooked beef patties by hydrolyzed potato protein is related to its reducing and radical scavenging ability. J. Agric. Food Chem, 2005(53): 9186-9192
8. Pena-Ramos E A, Xiong Y L. Antioxidative activity of whey protein hydrolysates in a liposomal system. J. Dairy Sci, 2001(84):2577-2583
9. Pena-Ramos E A, Xiong Y L, Arteaga G E. Fractionation and characterization for antioxidant activity of hydrolyzed whey protein. J. Sci. Food Agric, 2004(84):1908-1918
10. Zhu K X, Zhou H M, Qian H F. Antioxidant and free radical-scavenging activities of wheat germ protein hydrolysates (WGPH) prepared with alcalase. J. Process Biochem,2006(41): 1296-1302
11. Wang J S, Zhao M M, Zhao Q Z, et al. Antioxidant properties of papain hydrolysates of wheat gluten in different oxidation systems. J. Food Chem,2007 (101):1658-1663
12. Davalos A, Miguel M, Bartolome B, et al. Antioxidant activity of peptides derived from egg white proteins by enzymatic hydrolysis. J. Food Prot,2004(67):1939-1944
13. Mendis, E; Rajapakse, N; Byun, H.-G; et al. Investigation of jumbo squid (Dosidicus gigas) skin gelatin peptides for their in vitro antioxidant effects. J. Life Sci, 2005(77):2166-2178
14. Rajapakse N,Mendis E,Byun H G,et al. Purification and in vitro antioxidative effects of giant squid muscle peptides on free radical-mediated oxidative systems .J. Nutr Biochem, 2005, 16(9):562-569
15.夏向东,吕飞杰,台建祥.抗氧化剂的功效及体外抗氧化活性的评价[J].食品科学,2001,22(1):89-93
16.郑晶泉.抗氧化剂抗氧化实验研究进展[J].国外医学卫生学分册,2000,27(1):37-40
17.徐清萍,钟桂芳,孟君.抗氧化剂抗氧化方法研究进展[J].食品工程,2007(2):23-25
18. Block G, Patterson B H, Subar A F. Fruit, vegetables, and cancer prevention: a review of the epidemiological evidence. J. Nutr. Cancer, 1992 (18):11-29
19. Michel Soulié. What is the Role of Surgery for Locally Advanced Disease. J. European Urology Supplements, 2008(7):400-405
20.童荣生.自由基与心血管疾病[J].现代临床医学,2007,33(2):188-191
21. Willcox J K, Ash S, Catignani G. Antioxidants and prevention of chronic disease. J. Food Sci. Nutr, 2004 (44): 275-295
22.玄国东.米糟蛋白提取及酶法制备抗氧化活性肽及降血压活性肽的研究[D]:[博士学位论文],浙江:浙江大学,2005
23.吴亚梅,陈健,李维峰.玉米蛋白粉深加工应用的新进展[J].现代食品科技,2007,25(4): 97-100
24. Blanca Hernández-Ledesma,Lourdes Amigo, Isidra Recio, et al. ACE-inhibitory and radical-scavenging activity of peptides derived fromβ-lactoglobulin f(19-25). Interactions with ascorbic acid. J. Agric. Food Chem, 2007(55):3392-3397
25. Anne Pihlanto, Sari Akkanen, Hannu J. Korhonen. ACE-inhibitory and antioxidant properties of potato .J. Food Chemistry, 2008(109):104-112
26.张晓峰.腐乳发酵及其抗氧化和ACE抑制活性研究[D]:[博士学位论文].北京:中国农业大学,2006
27.郑喜群.玉米黄粉的酶解工艺与抗氧化活性肽的制备[D]:[博士学位论文].北京:中国农业大学, 2006
28. Shukla R, Cheryan M. Zein: the industrial protein from corn. J. Ind. Crops Prod,2001(13): 171–192
29.常亮,韩恩山,胡建修,等.玉米醇溶蛋白的研究及应用[J].化工中间体,2006(1):10-12
30.白艳菊,朱永军,代英杰,等.玉米醇溶蛋白的提取和应用[J].黑龙江农业科学,2001(1): 34-35
31陈石良,吴建平,谷文英.高F值寡肽的研究进展[J].食品与机械,1998(2):12-14
32.郭维静.玉米蛋白水解制取玉米肽的研究.[D]:[硕士学位论文],大连:大连理工大学
33.鲁晓翔,陈新华,唐津忠.酶法改性玉米蛋白功能特性的研究[J].食品科学,2000,21(12): 13-15
34.王遂,尤旭.微生物蛋白酶对玉米皮蛋白水解作用的研究[J].食品工业科技,2000,21(4): 22-24
35. Seiko Yano, Kazumasa Suzuki, Gunki Funatsu. lsolation from a-zein of themokysin peptides with Angiotensin1-converting Enzyme In hibitory Activity. J. Biosci. Biotech. Biochem,1996,60(4): 661-663
36.王梅,戴军.高效凝胶过滤色谱法测定玉米蛋白酶解物寡肽的分子量分布[J].食品与发酵工业,1998,24(5):25-27
37.王梅,谷文英.酶解玉米黄粉蛋白制备可溶性肽[J].粮油食品科技,1999,7(1):1-3
38.郑冬梅,孔保华,李升福.玉米蛋白及其水解肽的研究动态[J].食品与发酵工业,2002, 28(11):55-59
39.郑冬梅,李升福,孔保华.玉米蛋白水解条件的优化研究[J].食品科学,2002,23(3):52-56
40.赵金兰.用玉米渣生产降血压活性肽[J].食品与机械,1998(5):7-8
41.何慧,谢笔钧,杨卓.大豆蛋白和玉米蛋白酶解肽及其活性研究[J].粮油食品科技,2002,
10(1):14-16
42. Xu L, Zhang X Z, GUO Y, et al. A Study on Bioactivity of Corn Peptides with Low Molecular Weight Effect of an In take of them on Alcohol Metabolism in Rats. J. CHEM. RES. CH IN ESE U, 2003, 19(2): 180-182
43.李晶.玉米肽抗疲劳作用的实验研究[J].食品与机械, 2004, 20(1): 11-13
44.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(8):156-158
45.张强,阚国仕,陈红漫.玉米抗氧化肽的分离制备及其体外抗氧化活性的研究[J].中国粮油学报,2005,20(5):36-39
46.刘萍.酶解玉米蛋白提取ACE抑制肽的研究[D]:[硕士学位论文],江苏:江南大学,2006
47.吴炜亮,吴国杰,梁道双等. ACE抑制肽的生理功能和研究进展[J].现代食品技,2006, 22(3):251-254
48.罗郁中.台灣六種野生植物果實抗氧化功能評估[D]:[硕士学位论文],台湾:国立大,民国九十二年
49. [109]熊双丽,李安林,金征宇.猪喉软骨硫酸软骨素体外超氧阴离子自由基清除机理探讨[J].食品科学,2008,29(2):362-365
50.程云辉.麦胚蛋白酶解物的制备、结构及其生物活性功能的研究[D]:[博士学位论文].江苏:江南大学,2006
51.严群芳.大豆抗氧化肽的分离及其生物活性研究[D]:[硕士学位论文].江苏:南京农业大学,2006
52.张铁鹰,汪儆.单胃动物体外消化模拟技术研究进展[J].动物营养学报,2005,17(2):1-8
53.刘选珍,乐国伟,施用晖.饲料蛋白质体外消化率及其影响因素的研究[J].四川农业大学学报,1996(14):51-57
54. Arlene Wolzak, Ricardo Bressani, Roberto Gomez Brenes. A comparison of in vivo and in vitro estimates of protein digestibility of native and thermally processed vegetable proteins. J. Qual Plant Plant Foods ttum Nutr, 1981(31):31-43
55.王显生,杨晓泉,唐传核.大麻蛋白的营养评价[J].现代食品科技,2005,23(7):10-14
56.汤国营,戴军,蔡木易,等.大豆低聚肽分子质量分布的测定及体外消化实验[J].食品与发酵工业,2006,32(5):131-133
57. Cyrille Krul, Anja Luiten-Schuite, Aschwin Tenfelde, et al. Antimutagenic activity of green tea and black tea extracts studied in a dynamic in vitro gastrointestinal model. J. Mutation Research,2001(474):71-85
58. Blanca Hernández-Ledesma, Lourdes Amigo, Mercedes Ramos,et al. Release of angiotensin converting enzyme-inhibitory peptides by simulated gastrointestinal digestion of infant formulas. J. International Dairy Journal,2004(14):889-898
59. Blanca Hernández-Ledesma, Ana Quirós, Lourdes Amigo, et al. Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin. J. International Dairy Journal, 2007(17):42-49
60. Lo, W. M. Y; Li-Chan, E. C. Angiotensin I converting enzyme inhibitory peptides from in vitro pepsin-pancreatin digestion of soy protein. J. Agric. Food Chem, 2005(53):3369-3376
61. Marta Miguel, María J. Alonso, Mercedes Salaices, et al. Antihypertensive, ACE-inhibitory and vasodilator properties of an egg white hydrolysate: Effect of a simulated intestinal digestion. J. Food Chemistry, 2007 (104):163-168
62. Zhong-Ji Qian, Won-Kyo Jung, Hee-Guk Byun, et al. Protective effect of an antioxidative peptide purified from gastrointestinal digests of oyster, Crassostrea gigas against free radical induced DNA damage. J. Bioresource Technology, 2008(99):3365-3371
63.杨梅琳.蚕蛹蛋白的酶法水解及其产物的抗氧化性研究[D]:[硕士学位论文].江苏:江南大学,2006
64. Garrett, D. A; Failla, M. L; Sarama, R. J. Development of an in vitro digestive method to assess carotenoid bioavailability from meals. J. Agric. Food Chem, 1999(47):4301-4309
65. Adler-Nissen, J. Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. J. Agric. Food Chem, 1979(27):1256-1262
66. Spellman, D.; McEvoy, E.; O’Cuinn, G.; FitzGerald, R. J. Proteinase and exopeptidase hydrolysis of whey protein: Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis. J. Intl. Dairy, 2003(13):447-453
67.余冰宾,生物化学实验指导[M].第1版.北京:清华大学出版社,2004.133-136
68. Pellegrini, N. R; Proteggente, N; Pannala, A;et al. Antioxidant activity applying an improved ABTS radical cation decolorization assay. J. Free Radical Biol. Med, 1999(26):1231-1237
69. Wu, H.-C; Chen, H.-M; Shiau, C.-Y. Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus). J. Food Res. Intl, 2003(36): 949-957
70. Marcuse, R. Antioxidative effect of amino-acids. J. Nature, 1960(186):886-887
71. Saiga A, Tanabe S, Nishimura T. Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment. J. Agric. Food Chem, 2003(51):3661-3667
72. Decker, E. A; Crum, A. D; Calvert, J. T. Differences in the antioxidant mechanism of carnosine in the presence of copper and iron. J. Agric. Food Chem, 1992(40):756-759
73.布冠好,杨国宇,李宏基等.肌肽对猪肉匀浆的氧化抑制作用研究[J].河南工业大学学报(自然科学版),2006,27(3):62-65
74. Chen H M, Muramoto K, Yamauchi F. Structural analysis of antioxidative peptides from soybeanβ-conglycinin. J. Agric. Food Chem, 1995(43):574-578.
75.豆康宁.酶解大豆分离蛋白制取大豆肽及其抗氧化性研究[D]:[硕士学位论文].河南:河南工业大学,2007
76.姜瞻梅.酶解酪蛋白生产ACE抑制肽[D]:[硕士学位论文].哈尔滨:东北农业大学,2003
77.张宇昊.花生短肽制备及其功能活性研究[D]:[博士学位论文].北京:中国农业科学院. 2007
78.倪莉,陶冠军,戴军,等.血管紧张素转化酶活性抑制剂丝素肽的分离、纯化和结构鉴定[J].色谱,2001,19(3):222-225
79.于娅,杨瑞金,王璋.牡蛎功能短肽的制备及ACE抑制活性[J].无锡轻工大学学报,2004,23(2):49-52
80.管骁姚惠源.酶法制备燕麦麸蛋白ACE抑制肽的研究.食品与机械,2006, 22(6):12-15
81.刘志国,吴琼,吕玲肖,等.酶解米糠蛋白分离提取ACE抑制肽及其结构研究[J].食品科学, 2007,28(3):223-227
82.吴炜亮,吴国杰,梁道双等.ACE抑制肽的生理功能和研究进展[J].现代食品技,2006,22(3):251-254
83.任维栋.猪骨胶原蛋白酶解物中血管紧张素转换酶抑制剂的提纯[J].生物化学杂志,1996,l2(6):693-697
84.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(8):156-158
85.张莉莉,严群芳,王恬.大豆生物活性肽的分离及其抗氧化活性研究[J].食品科学,2007, 28(5):208-211
86.徐力,赵忠岩,李鸿梅等.大豆蛋白的小分子酶解产物抗氧化活性研究.吉林农业大学学报, 2007,29(1):48-52
87.郭红,李茂辉,阎萍,等.大豆小分子肽抗氧化效应的初步研究[J].中华医学研究杂志,2007,7(3):196-198
88. Chen H M, Muramoto K, Yamauchi F, et al. Antioxidant activity of designed peptides based on the antioxidative peptide isolated from digests of a soybean protein. J. Agric. Food Chem, 1996(44):2619-2623
89. JE J Y,PARK P J,KIM S K. Antioxidant activity of a peptide isolated from alaska pollack (Theragra chalcogramma) frame protein hydrolysate.J.Food Res Int, 2005,38(1):45-50
90.徐明生.鸡蛋卵白蛋白酶解物抗氧化肽研究[D]:[博士学位论文].陕西:陕西师范大学, 2006