超声波协同双酶复合酶解米渣蛋白制备ACE抑制肽工艺研究
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摘要
运用超声波协同双酶复合酶法水解米渣蛋白制备ACE抑制肽。超声波预处理后米渣蛋白水解物ACE抑制活性显著上升,碱性蛋白酶水解产物ACE抑制活性最强。通过单因素分析和响应面优化,得出最优水解条件为:超声功率1 000W,超声时间25min,酶解时间2.5h,料液比1∶8,加酶量3 000U/g。在此基础上复合中性蛋白酶水解,水解时间缩减至2.0h。水解产物通过超滤以及Sephadex G-25凝胶层析后,得到一分子量为338u,最强ACE抑制活性IC_(50)为116μg/ml组分P2。
Double enzyme hydrolysis assisted with ultrasonic was used to hydrolyze rice residue protein(RRP)to produce angiotensinⅠ-converting enzyme(ACE)inhibitory peptides.The ACE inhibitory effect was significantly improved when the RRP was pretreated with ultrasonic,and the ACE inhibitory activity of hydrolysates by alcalase was the highest.The hydrolysis conditions optimized by single factor experiment and response surface method were as follows:ultrasonic power 1 000 W,ultrasonic time 25 min,enzymatic hydrolysis time 2.5 h,the solid to liquid ratio 1∶8,and enzyme concentration 3 000 U/g.Pre-hydrolyzing RRP with neutrase resulted in the reduction of hydrolysis time to 2.0 h.The hydrolysate was then separated by ultrafiltration and Sephadex G-25 gel filtration.ACE inhibitory peptide(P2),which showed the highest ACE inhibitory activity(IC_(50)=116μg/ml)and a molecular weight of 338 u,was isolated.
Double enzyme hydrolysis assisted with ultrasonic was used to hydrolyze rice residue protein(RRP)to produce angiotensinⅠ-converting enzyme(ACE)inhibitory peptides.The ACE inhibitory effect was significantly improved when the RRP was pretreated with ultrasonic,and the ACE inhibitory activity of hydrolysates by alcalase was the highest.The hydrolysis conditions optimized by single factor experiment and response surface method were as follows:ultrasonic power 1 000 W,ultrasonic time 25 min,enzymatic hydrolysis time 2.5 h,the solid to liquid ratio 1∶8,and enzyme concentration 3 000 U/g.Pre-hydrolyzing RRP with neutrase resulted in the reduction of hydrolysis time to 2.0 h.The hydrolysate was then separated by ultrafiltration and Sephadex G-25 gel filtration.ACE inhibitory peptide(P2),which showed the highest ACE inhibitory activity(IC_(50)=116μg/ml)and a molecular weight of 338 u,was isolated.
引文
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[22] WU J P,ROTIMI E A,SHURYONAKAI.Structural requirements of angiotensinⅠ-converting enzyme inhibitory peptides:quantitative structure-activity relationship study of diand tripeptides[J].Journal of Agricultural and Food Chemistry,2006,54(3):732-738.
[23] CHEN J W,LIU S S,YE R,et al.AngiotensinⅠ-converting enzyme(ACE)inhibitory tripeptides from rice protein hydrolysate:Purification and characterization[J].Journal of Functional Foods,2013(5):1 684-1 692.
[24] WANG X M,CHEN H X,FU X G,et al.A novel antioxidant and ACE inhibitory peptide from rice bran protein:Biochemical characterization and molecular docking study[J].LWTFood Science and Technology,2017,75:93-99.
[2] CHRASTIL J,ZARINS Z M.Influence of storage on peptide subunit composition of rice oyrzenin[J].J Agric Food Chem,1992,40(6):927-930.
[3] ZARINS Z,CHRASTIL J.Separation and purification of rice oyrzenin subunits by anion exchange and gel permeation chromatography[J].J Agric Food Chem,1992,40(9):1 599-1 601.
[4]王章存,刘卫东,申瑞玲,等.大米蛋白水解机制研究——Ⅲ酶水解对大米蛋白-糖结合特性的影响[J].中国粮油学报,2007,22(6):5-7.
[5] SHIN Z I,AHN C W,NAM H S,et al.Fractionation of angiotensin converting enzyme(ACE)inhibitory peptides soybean paste[J].Korean J Food Sc Technol,1995,27:230-234.
[6] DASKAYA-DIKMEN C,YUCETEPE A,KARBANCIOGLUGULER F,et al.AngiotensinⅠ-converting enzyme(ace)-inhibitory peptides from plants[J].Nutrients,2017(9):1-19.
[7] HE H L,LIU D,MA C B.Review on the angiotensinⅠ-converting enzyme(ACE)inhibitor peptides from marine proteins[J].Applied Biochemistry and Biotechnology,2013(3):738-749.
[8] HISHAM R I,AHMED S A,TAKESHI M.Novel angiotensinⅠ-converting enzyme inhibitory peptides from caseins and whey proteins of goat milk[J].Jornal of Advanced research,2017,8(1):63-71.
[9] LUCA A,JONATHAN O,ALAN L K,et al.Composition and protein profile analysis of rice protein ingredients[J].Journal of Food Composition and Analysis,2017,59:18-26.
[10] FANG H,LUO M,SHENG Y,et al.The antihypertensive effect of peptides:A novel alternative to drugs[J].Peptides,2008,29(6):1 062–1 071.
[11]贾俊强,马海乐,曲文娟,等.超声预处理大米蛋白制备抗氧化肽[J].农业工程学报,2008,24(8):288-293.
[12]王申,周佳,徐晶,等.超声、微波联合预处理大米蛋白制备ACE抑制肽工业优化[J].食品与机械,2014,36(3):159-162.
[13]魏康,李云亮,杨雪,等.逆流超声辅助酶解法制备大米ACE抑制肽[J].食品工业科技,2018,39(4):100-105.
[14] YAN Q J,HUANG L H,SUN Q,et al.Isolation,identification and synthesis of four novel antioxidant peptides from rice residue protein hydrolyzed by multiple proteases[J].Food Chemistry,2015,179:290-295.
[15] ADLER-NISSEN J.Enzymic hydrolysis of food proteins[M].Amsterdam:Elsevier Applied Science Publisher,1986.
[16]李汉洋,李建杰,王帅,等.核桃多肽的抗氧化活性及其分子量、氨基酸组成特性研究[J].食品工业科技,2018,39(13):1-7.
[17]阎震,郭淑,张金宝,等.酶解牡丹籽粕蛋白制备抗氧化肽的工艺优化[J].食品工业科技,2018,39(7):168-174.
[18]张龙翔,张庭芳,李令媛.生化实验方法和技术[M].北京:高等教育出版社,1981:124-132.
[19] CUSHMAN D W,CHEUNG H S.Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung[J].Biochemistry Pharmacology,1971,20:1 637-1 648.
[20] LI S Y,YANG X,ZHANG Y Y,et al.Effects of ultrasound and ultrasound assited alkaline pretrements on the enzymolysis and structural characteristic of rice protein[J].UltrasonicSonochemistry,2016,31:20-28.
[21] YAO J,CHEN Q S,GONG S S.Research status of probioctics antihypertensive peptides and trends of domestic and international new production development[J].Journal of Food Science,2007(9):590-594.
[22] WU J P,ROTIMI E A,SHURYONAKAI.Structural requirements of angiotensinⅠ-converting enzyme inhibitory peptides:quantitative structure-activity relationship study of diand tripeptides[J].Journal of Agricultural and Food Chemistry,2006,54(3):732-738.
[23] CHEN J W,LIU S S,YE R,et al.AngiotensinⅠ-converting enzyme(ACE)inhibitory tripeptides from rice protein hydrolysate:Purification and characterization[J].Journal of Functional Foods,2013(5):1 684-1 692.
[24] WANG X M,CHEN H X,FU X G,et al.A novel antioxidant and ACE inhibitory peptide from rice bran protein:Biochemical characterization and molecular docking study[J].LWTFood Science and Technology,2017,75:93-99.