ACE- inhibitory peptides derived from milk protein by continuous coupling of enzymatic hydrolysis and membrane separation without material feeding
-
摘要: 为了克服传统酶解技术的不足,提高酶解反应效率,采用不补料酶膜耦合反应制备牛乳蛋白ACE抑制肽。考察超滤膜对牛乳蛋白及酶的截留、膜通量及滤出液的ACE抑制率和IC50的影响,确定出超滤膜最佳的截留分子量为5000 u;研究了反应时间、底物浓度、加酶量和循环泵转速对蛋白转化率的影响,确定最优酶解工艺条件:底物浓度7%（W/W）、加酶量2000 U/g、反应时间100 min、循环泵转速100 r/min,该条件下蛋白转化率、单位酶产肽量、ACE抑制率分别达到53.16%、39.59 g肽/g酶、76.81%,与对照组相比蛋白转化率、单位酶产肽量、ACE抑制率分别提高了16.99%、16.61%、18.55%;采用高效凝胶过滤色谱法测定超滤液中ACE抑制肽的分子量分布,发现样品中分子量≤2253 u组分质量分数为94.62%,超滤膜对截留分子量大小控制准确。因此不补料酶膜耦合反应可为酶法制备牛乳蛋白ACE抑制肽提供一种更为高效的方法。Abstract: To overcome disadvantage of traditional enzymatic hydrolysis and improve reaction effect,ACE- inhibitory peptides from milk protein were prepared by coupling of enzymatic hydrolysis and membrane separation.According to the comparison of protein retention,protease retention,membrane flux,permeates' ACE- inhibitory activity and IC50,the molecular weight of ultra-filtration membrane was determined as 5000 u.The effects of reaction time,substrate concentration,enzymatic dosage and cycle pump speed on conversion ratio of protein were investigated respectively.The optimal conditions obtained from the single factor tests were: substrate concentration7%( W/ W),enzymatic dosage 2000 U / g,reaction time 100 min and cycle pump speed 100 r / min. Under the optimal conditions,conversion ratio of protein reached 53.16% with productivity of 39.59 g peptide·( g protease)-1 and the inhibitory rate of ACE was 76.81%.Compared with the control group,the three contents were increased by16.99%,16.61% and 18.55% respectively. The relative molecular weight of permeates was measured by high performance size exclusion chromatography( HPSEC). The results showed that the molecular weight could be effectively controlled by ultra- filtration and 94.62% of sample was less than 2253 u. Therefore,it is suggested that the reaction of coupling of enzymatic hydrolysis and membrane separation can provide a more effective method for the preparation of ACE- inhibitory peptides from milk protein.
-
[1] 于晓庆,王雄,司佳.酶法制备乳源ACE抑制肽[J].食品研究与开发,2011,32(9):134-136. [2] Fitzgerald R J,Meisel H.Milk protein-derived peptide inhibitors of angiotensin-I-converting enzyme[J].British Journal of Nutrition,2000,84:33-37.
[3] 洪伟.酪蛋白中血管紧张素转换酶(ACE)抑制肽的研究[D].芜湖:安徽工程大学,2010. [4] 魏春燕.酪蛋白非磷肽酶膜耦合法制备ACE抑制肽[D].无锡:江南大学,2011. [5] 崔洁,孔祥珍,周惠明,等.小麦面筋蛋白预处理及酶膜耦合连续反应研究[J].食品工业科技,2011,32(6):188-191. [6] 熊治清,徐志宏,魏振承,等.酶膜反应器在蛋白酶解过程中的研究进展[J].食品科技,2010,35(1):88-92. [7] QU Wenjuan,MA Haile,ZHAO Weirui,et al.ACE-inhibitory peptides production from defatted wheat germ protein by continuous coupling of enzymatic hydrolysis and membrane separation:Modeling and experimental studies[J].Chemical Engineering Journal,2013,226:139-145.
[8] 丁青芝,马海乐,骆琳,等.酶解-膜分离耦合技术制备米糠蛋白活性多肽的研究[J].高校化学工程学报,2009,23(4):632-638. [9] 姜瞻梅,田波,吴刚,等.酶解牛乳酪蛋白制备ACE抑制肽的研究[J].中国食品学报,2007,7(6):39-43. [10] Seronei C C,ZHANG Wang,ZHANG Wang.Hydrolysis of whey protein isolate in a tangential flowfilter membrane reactorⅡ[J].Journal of Membrane Science,2006,286:322-332.
[11] 姚喜梅.连续式酶膜耦合反应制备魔芋葡甘露寡糖[D].北京:中国农业科学院,2011. [12] 齐崴,何志敏,何明霞.酶解反应与膜分离耦合连续制备酪蛋白磷酸肽[J].化学工程,2006,34(4):43-46. [13] 刘妍妍,张丽萍.牛乳酪蛋白源降血压肽的分离及其分子量分布研究[J].中国乳品工业,2006,34(5):26-28. [14] 徐鑫,蔡丽丽,刘国艳,等.响应曲面法优化酶解条件制备乳源ACE抑制肽[J].食品科学,2012,33(5):208-212. [15] 张艳,胡志和,闫星,等.胃蛋白酶水解酪蛋白制备ACE抑制肽的条件[J].食品科学,2010,31(14):42-46. [16] OTTE J,SHALABY S M,ZAKORA M,et al.Angiotensin converting enzyme inhibitory activity of milk protein hydrolysates:Effect of substrate,enzyme and time of hydrolysis[J].International Dairy Journal,2007,17:488-503.
[17] 陈钧辉,陶立,李俊,等.生物化学实验[M].北京:科学出版社,2003:59-61. [18] 吴琼英,马海乐,骆琳,等.高效液相色谱法测定血管紧张素转化酶抑制剂的活性[J].色谱,2005,23(1):79-81. [19] 陈耿华,王磊,王强,等.用于连续制备乳清蛋白ACE抑制肽的酶膜反应器运行条件优化[J].农产品加工,2011(9):4-7. [20] YANG Sen,DING Wenyong,CHEN Hongzhang.Enzymatic hydrolysis of corn stalk in a hollow fiber ultrafiltration membrane reactor[J].Biomass and Bioenergy,2009,33:332-336.
[21] 黄文浩.连续化酶膜耦合法制备高活性ACE抑制玉米肽研究[D].武汉:华中农业大学,2011. [22] WU Shufen,QI Wei,LI Tonghe,et al.Simultaneous production of multi-functional peptides by pancreatic hydrolysis of bovine casein in an enzymatic membrane reactor via combinational chromatography[J].Food Chemistry,2013,141:2944-2951.
[23] 蒋菁莉,任发政,蔡华伟.牛乳酪蛋白降血压肽的超滤分离[J].食品科学,2006,27(7):124-128. [24] 夏镇波.酪蛋白源血管紧张素转化酶抑制肽制备及分离纯化研究[D].南京:南京农业大学,2008. [25] 李文,马海乐,王金斌.酶解反应与膜分离耦合连续制备紫菜降血压肽[J].食品开发与机械,2008(10):71-75. -
期刊类型引用(8)
1. 高扬，李家磊，严松，管立军，王崑仑，李波，周野，姚鑫淼，陈凯新，任传英，卢淑雯. 低温高剪切挤压重组制备工程糙米工艺优化及其品质特性分析. 中国粮油学报. 2025(01): 23-32 . 
百度学术
2. 汪佳琦，刘静雪，张力. 挤出处理对糙米的营养特性研究进展. 粮食问题研究. 2023(01): 41-44 . 百度学术
3. 李甜，丁志刚，陈雅南，陈磊. 富铬发芽糙米粉性质分析及其在馒头中的应用研究. 食品安全质量检测学报. 2023(11): 121-129 . 百度学术
4. 林华杏，覃笛根，陈强，孟晓雪，郑煜东，谭北平，迟淑艳. 挤压膨化对大豆浓缩蛋白物理特性和蛋白组分的影响. 饲料工业. 2023(19): 72-77 . 百度学术
5. 李斌，庄坤，丁文平，王国珍. 制粉工艺对糯糙米粉理化特性的影响. 食品科技. 2022(09): 125-130 . 百度学术
6. 方浩标，郑经绍，余宏达，许立益，陆梓洋，黄苇. 挤压膨化对紫糙米粉营养品质及理化性质的影响. 食品工业科技. 2021(19): 70-77 . 本站查看
7. 关桦楠，吴永存，张娜. 糙米蒸煮品质改良加工技术研究进展. 粮食与油脂. 2021(12): 26-28+40 . 百度学术
8. 戚明明，郭鹏，汪陈洁，董爽，马成业. 挤压对淀粉微观结构和理化性质影响的研究进展. 食品工业科技. 2020(05): 305-310 . 本站查看
其他类型引用(13)
计量
- 文章访问数:
- HTML全文浏览量:
- PDF下载量:
- 被引次数: 21
下载:
