卵白蛋白低聚肽的体外抗氧化及ACE抑制作用
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摘要
以卵白蛋白低聚肽为研究对象,测定了理化成分,分子质量分布和氨基酸组成基础之上;通过DPPH·法、邻苯三酚自氧化法和水杨酸钠法测定对DPPH自由基(DPPH·)、超氧阴离子自由基(O_2~-·)和羟自由基(·OH)的清除能力,分析了其体外抗氧化作用;另外通过反向高效液相色谱法测定了对血管紧张素转化酶(angiotensin converting enzyme,ACE)的抑制作用。结果显示,分子质量低于1 000 u的卵白蛋白低聚肽占总蛋白的92.58%,并主要集中在140～500 u(占71.24%);体外抗氧化实验中,抗氧化能力与质量浓度呈现出良好的剂量关系,DPPH·、O_2~-·和·OH的半抑制浓度(IC_(50))分别为0.25、25、0.64 mg/m L;另外卵白蛋白低聚肽显示出较好的ACE抑制作用,IC_(50)值为1.5 mg/m L。结果表明,卵白蛋白低聚肽具有良好的抗氧化和ACE抑制作用,为相关抗氧化和降血压等保健类食品的开发利用提供一定的理论依据。
The ovalbumin oligopeptides were used as research object,and their physicochemical components,molecular weight distribution and amino acid composition were determined.In order to analyze their antioxidant activities in vitro,their abilities to scavenge DPPH free radicals,superoxide anion radicals and hydroxyl radicals were determined.In addition,their effects on inhibiting angiotensin converting enzyme(ACE)were analyzed by reverse high performance liquid chromatography.The results showed that the ovalbumin oligopeptides with a molecular weight of less than 1 000 u accounted for 92.58%of the total protein,and were mainly concentrated between 140 u and 500 u(71.24%).Moreover,their antioxidant capacities showed good dose relationships with their mass concentration.Their semi-inhibitory concentrations(IC_(50))of DPPH free radicals,superoxide anion radicals and hydroxyl radicals were 0.25 mg/m L,25 mg/m L,and 0.64 mg/m L,respectively.Furthermore,ovalbumin oligopeptides showed better ACE inhibitory effects with an IC_(50)value of 1.5 mg/m L.The results showed that ovalbumin oligopeptides have good antioxidative and ACE inhibition effects,which provides a theoretical basis for developing and utilizing health-care foods with antioxidative and blood pressure lowering effects.
The ovalbumin oligopeptides were used as research object,and their physicochemical components,molecular weight distribution and amino acid composition were determined.In order to analyze their antioxidant activities in vitro,their abilities to scavenge DPPH free radicals,superoxide anion radicals and hydroxyl radicals were determined.In addition,their effects on inhibiting angiotensin converting enzyme(ACE)were analyzed by reverse high performance liquid chromatography.The results showed that the ovalbumin oligopeptides with a molecular weight of less than 1 000 u accounted for 92.58%of the total protein,and were mainly concentrated between 140 u and 500 u(71.24%).Moreover,their antioxidant capacities showed good dose relationships with their mass concentration.Their semi-inhibitory concentrations(IC_(50))of DPPH free radicals,superoxide anion radicals and hydroxyl radicals were 0.25 mg/m L,25 mg/m L,and 0.64 mg/m L,respectively.Furthermore,ovalbumin oligopeptides showed better ACE inhibitory effects with an IC_(50)value of 1.5 mg/m L.The results showed that ovalbumin oligopeptides have good antioxidative and ACE inhibition effects,which provides a theoretical basis for developing and utilizing health-care foods with antioxidative and blood pressure lowering effects.
引文
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[21]黄薪蓓,许庆彪,刘建新.肠道氨基酸和小肽转运载体的基因表达、影响因素与分子调控机制[J].动物营养学报,2015,27(1):21-27.
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[23]彭孟圆,仓静,方芳.谷氨酸转运体-1相关核转录因子及其药物调控的研究进展[J].临床麻醉学杂志,2017,33(4):408-411.
[24]姜廷波,王莹,赵艳飞,等.肠道中赖氨酸转运载体的研究进展[J].中国畜牧杂志,2017,53(1):7-11.
[25]王良峡,刘若颖,林福鸿,等.支链氨基酸在运动中的作用研究进展[J].生物资源,2015,37(1):7-12.
[26]陈冠群.机体总抗氧化力的氨基酸和基因基础以及氧化对抗体活性及其它实验室指标的影响[D].大连:大连医科大学,2014.
[27]布冠好,杨国宇,李宏基,等.新型抗氧化物质---组氨酸二肽的研究现况和展望[J].中国动物保健,2005(6):28-30.
[28]KONG Baohua,Youlin Xiong.Antioxiunt activity of zein hydrolysates in a liposome system and the possible mode of action[J].J Agric Food Chem,2006,54(16):6 059-6 068.
[29]丁苗.发酵酸肉中ACE抑制肽的分离纯化、特性及其对血管内皮细胞功能因子的影响研究[D].重庆:西南大学,2015.
[30]付琴琴,王岸娜,吴立根.食醋抗氧化活性的研究进展[J].河南工业大学学报(自然科学版),2012,33(1):88-93.
[31]王宁丽,徐月敏,刘毅,等.鱼皮低聚肽氨基酸组成及抗氧化活性研究[J].食品研究与开发,2016,37(2):31-35.
[32]GLIGOROVSKI S,STREKOWSKI R,BARBATI S,et al.Environmental implications of hydroxyl radicals(·OH)[J].Chemical Reviews,2015,115(24):13 051-13 092.
[33]王福海,黄成华.活性氧自由基的研究进展[J].广州化工,2013,41(16):10-12.
[34]RAJAPAKSE N,MENDIS E,JUNG W K,et al.Purification of a radical scavenging peptide from fermented mussel sauce and its antioxiunt properties[J].Food Research International,2005,38(2):175-182.
[35]BOUGATEF A,NEDJAR-ARROUME N,RAVALLEC-PLR,et al.Angiotensin I-converting enzyme(ACE)inhibitory activities of sardinelle(Sardinella aurita)byproducts protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases[J].Food Chemistry,2008,111(2):350-356.
[36]刘文颖,林峰,金振涛,等.深海鲑鱼皮来源ACE抑制肽的分离及鉴定[J].现代食品科技,2016,32(6):170-176.
[37]HE H L,CHEN X L,WU H,et al.High throughput and rapid screening of marine protein hydrolysates enriched in peptides with angiotensin-I-converting enzyme inhibitory activity by capillary electrophoresis[J].Bioresource Technology,2007,98(18):3 499-3 505.
[38]翟学超,钟立成.鹿血抗氧化肽不同分子量肽段抗氧化活性的研究[J].经济动物学报,2015,19(3):144-146.
[39]ZHANG C H,CAO W H,HONG P Z,et al.Angiotensin I-converting enzyme inhibitory activity of peptic hydrolysate and its antihypertensive effect in spontaneously hypertensive rats[J].International Journal of Food Science&Technology,2010,44(10):2 042-2 048.
[40]WILSON J,HAYES M,CARNEY B.Angiotensin-I-converting enzyme and prolyl endopeptiuse inhibitory peptides from natural sources with a focus on marine processing byproducts[J].Food Chemistry,2011,129(2):235-244.
[41]左琦,吴秉宇,张建华,等.特异性蛋白酶酶解虾副产物制备ACE抑制肽[J].食品工业科技,2014,35(10):181-185.
[2]SUN C,SHEN W C,TU J,et al.Interaction between cell-penetrating peptides and acid-sensitive anionic oligopeptides as a model for the design of targeted drug carriers[J].Molecular Pharmaceutics,2014,11(5):1 583-1 590.
[3]潘兴昌,蔡木易.食源性低聚肽与特殊膳食食品的开发[C]//中国营养学会会员代表大会,2008.
[4]杨柳.含硫钠盐/钾盐抗氧化和清除自由基的活性研究及不同抗氧化剂配伍对抗氧化能力的影响[D].南宁:广西医科大学,2015.
[5]刘文颖,谷瑞增,鲁军,等.大豆低聚肽中抗氧化肽的分离纯化及结构鉴定[J].食品与发酵工业,2017,43(2):44-48.
[6]TOOPCHAM T,ROYTRAKUL S,YONGSAWATDIGULJ.Characterization and identification of angiotensin I-converting enzyme(ACE)inhibitory peptides derived from tilapia using Virgibacillus halodenitrificans SK1-3-7 proteinases[J].Journal of Functional Foods,2015,14:435-444.
[7]PIHLANTO A,JOHANSSON T,MKINEN S.Inhibition of angiotensin I-converting enzyme and lipid peroxiution by fermented rapeseed and flaxseed meal[J].Engineering in Life Sciences,2012,12(4):450-456.
[8]卫生部食品卫生监督检验所.GB/T 5009.5-2003食品中蛋白质的测定[S].北京:中国标准出版社,2003.
[9]中国食品发酵工业研究院.GB/T 22729-2008海洋鱼低聚肽粉[S].北京:中国标准出版社,2003.
[10]卫生部食品卫生监督检验所.GB/T 5009.6-2003食品中脂肪的测定[S].北京:中国标准出版社,2003.
[11]卫生部食品卫生监督检验所.GB/T 5009.4-2003食品中灰分的测定[S].北京:中国标准出版社,2003.
[12]卫生部食品卫生监督检验所.GB/T 5009.3-2003食品中水分的测定[S].北京:中国标准出版社,2003.
[13]卫生部食品卫生监督检验所.GB/T 5009.124-2003食品中氨基酸的测定[S].北京:中国标准出版社,2003.
[14]黄开华,高新华,陈伟,等.鹿血抗氧化活性肽分子量测定及其氨基酸组成分析[J].应用技术学报,2017,17(1):37-40.
[15]刘文颖,徐姗姗,谷瑞增,等.海洋胶原低聚肽中抗氧化肽的分离及鉴定[J].食品工业科技,2017,38(6):101-105.
[16]张强,辛秀兰,杨富民,等.红树莓果醋酿造过程中抗氧化性能的变化[J].食品科学,2016,37(3):6-11.
[17]席利莎,孙红男,木泰华,等.甘薯茎叶多酚的体外抗氧化活性与加工稳定性研究[J].中国食品学报,2015,15(10):147-156.
[18]WANG Jiapei,HU Jianen,CUI Jinzhe,et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats[J].Food Chemistry,2008,111(2):302-308.
[19]刘文颖,林峰,金振涛,等.玉米低聚肽的体外抗氧化作用[J].食品科学,2011,32(5):22-26.
[20]刘文颖,谷瑞增,鲁军,等.小麦低聚肽中抗氧化肽的分离纯化及结构鉴定[J].食品工业,2016(6):106-108.
[21]黄薪蓓,许庆彪,刘建新.肠道氨基酸和小肽转运载体的基因表达、影响因素与分子调控机制[J].动物营养学报,2015,27(1):21-27.
[22]LUNDQUIST P,ARTURSSON P.Oral absorption of peptides and nanoparticles across the human intestine:Opportunities,limitations and studies in human tissues[J].Adv Drug Deliv Rev,2016,106(Pt B):256-276.
[23]彭孟圆,仓静,方芳.谷氨酸转运体-1相关核转录因子及其药物调控的研究进展[J].临床麻醉学杂志,2017,33(4):408-411.
[24]姜廷波,王莹,赵艳飞,等.肠道中赖氨酸转运载体的研究进展[J].中国畜牧杂志,2017,53(1):7-11.
[25]王良峡,刘若颖,林福鸿,等.支链氨基酸在运动中的作用研究进展[J].生物资源,2015,37(1):7-12.
[26]陈冠群.机体总抗氧化力的氨基酸和基因基础以及氧化对抗体活性及其它实验室指标的影响[D].大连:大连医科大学,2014.
[27]布冠好,杨国宇,李宏基,等.新型抗氧化物质---组氨酸二肽的研究现况和展望[J].中国动物保健,2005(6):28-30.
[28]KONG Baohua,Youlin Xiong.Antioxiunt activity of zein hydrolysates in a liposome system and the possible mode of action[J].J Agric Food Chem,2006,54(16):6 059-6 068.
[29]丁苗.发酵酸肉中ACE抑制肽的分离纯化、特性及其对血管内皮细胞功能因子的影响研究[D].重庆:西南大学,2015.
[30]付琴琴,王岸娜,吴立根.食醋抗氧化活性的研究进展[J].河南工业大学学报(自然科学版),2012,33(1):88-93.
[31]王宁丽,徐月敏,刘毅,等.鱼皮低聚肽氨基酸组成及抗氧化活性研究[J].食品研究与开发,2016,37(2):31-35.
[32]GLIGOROVSKI S,STREKOWSKI R,BARBATI S,et al.Environmental implications of hydroxyl radicals(·OH)[J].Chemical Reviews,2015,115(24):13 051-13 092.
[33]王福海,黄成华.活性氧自由基的研究进展[J].广州化工,2013,41(16):10-12.
[34]RAJAPAKSE N,MENDIS E,JUNG W K,et al.Purification of a radical scavenging peptide from fermented mussel sauce and its antioxiunt properties[J].Food Research International,2005,38(2):175-182.
[35]BOUGATEF A,NEDJAR-ARROUME N,RAVALLEC-PLR,et al.Angiotensin I-converting enzyme(ACE)inhibitory activities of sardinelle(Sardinella aurita)byproducts protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases[J].Food Chemistry,2008,111(2):350-356.
[36]刘文颖,林峰,金振涛,等.深海鲑鱼皮来源ACE抑制肽的分离及鉴定[J].现代食品科技,2016,32(6):170-176.
[37]HE H L,CHEN X L,WU H,et al.High throughput and rapid screening of marine protein hydrolysates enriched in peptides with angiotensin-I-converting enzyme inhibitory activity by capillary electrophoresis[J].Bioresource Technology,2007,98(18):3 499-3 505.
[38]翟学超,钟立成.鹿血抗氧化肽不同分子量肽段抗氧化活性的研究[J].经济动物学报,2015,19(3):144-146.
[39]ZHANG C H,CAO W H,HONG P Z,et al.Angiotensin I-converting enzyme inhibitory activity of peptic hydrolysate and its antihypertensive effect in spontaneously hypertensive rats[J].International Journal of Food Science&Technology,2010,44(10):2 042-2 048.
[40]WILSON J,HAYES M,CARNEY B.Angiotensin-I-converting enzyme and prolyl endopeptiuse inhibitory peptides from natural sources with a focus on marine processing byproducts[J].Food Chemistry,2011,129(2):235-244.
[41]左琦,吴秉宇,张建华,等.特异性蛋白酶酶解虾副产物制备ACE抑制肽[J].食品工业科技,2014,35(10):181-185.
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