乳源锌螯合肽酶解制备及其螯合特性分析
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摘要
为研究乳源锌螯合肽酶解制备最佳工艺条件及其螯合特性,从而为促锌吸收活性物质的研究提供基础,以牛乳酪蛋白为原料,以锌螯合率为指标,确定胰蛋白酶酶解制备锌螯合肽的最佳工艺条件,并对最优条件下酶解物的分子质量分布及氨基酸组成进行测定,同时通过傅里叶变换红外扫描和紫外扫描手段对酶解物和肽锌复合物进行结构表征对比分析。结果显示,最佳酶解条件为时间3 h,温度50℃,p H 8. 5,底物质量浓度为70 g/L,加酶量0. 6%(质量分数)。最佳条件下,酶解物的锌螯合率为87. 78%,分子肽分子质量小,人体必需氨基酸含量丰富。螯合前后光谱存在明显差异,Zn~(2+)与羧基等进行了配位结合。乳源锌螯合肽具有较高的锌螯合活性,实验结果为补锌剂的生产应用奠定了基础。
This research aimed to study the condition for enzymatically preparing zinc chelating peptides from dairy sources and to illustrate their chelating characteristics. Trypsin was used for enzymolysis,and the molecular weight distribution and amino acid composition of the hydrolysates,as well as their structures and peptide zinc compounds were determined under the optimal preparation condition. The results showed that the optimal hydrolysis condition was at 50 ℃ and p H = 8. 5 for 3 h with 70 g/L casein and 0. 6% trypsin. Under the optimized condition,the zinc chelating rates of hydrolysates were 87. 78%. Moreover,the hydrolysates had small molecular peptides and were rich in essential amino acids. Furthermore,significant changes in spectra were observed before and after chelation,as Zn2 +could coordinate with carboxyl and other groups of peptides. In conclusion,dairy zinc chelating peptides have high zinc chelating activity. This study lays a foundation for producing and applying zinc supplements,and also for further studies on promoted absorption of active substances by zinc.
This research aimed to study the condition for enzymatically preparing zinc chelating peptides from dairy sources and to illustrate their chelating characteristics. Trypsin was used for enzymolysis,and the molecular weight distribution and amino acid composition of the hydrolysates,as well as their structures and peptide zinc compounds were determined under the optimal preparation condition. The results showed that the optimal hydrolysis condition was at 50 ℃ and p H = 8. 5 for 3 h with 70 g/L casein and 0. 6% trypsin. Under the optimized condition,the zinc chelating rates of hydrolysates were 87. 78%. Moreover,the hydrolysates had small molecular peptides and were rich in essential amino acids. Furthermore,significant changes in spectra were observed before and after chelation,as Zn2 +could coordinate with carboxyl and other groups of peptides. In conclusion,dairy zinc chelating peptides have high zinc chelating activity. This study lays a foundation for producing and applying zinc supplements,and also for further studies on promoted absorption of active substances by zinc.
引文
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[17]杨叶昆,李雪梅,唐自文,等.酪蛋白的酸水解及其棕色化反应产物在卷烟加香中的应用[J].烟草科技,2001(1):8-10.
[18]徐红梅,左凌波,高立娣.胰蛋白酶酶解酪蛋白工艺条件的优化[J].化工时刊,2015,29(3):1-5.
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[20]王晓萍.小麦胚芽蛋白源锌螯合肽的分离纯化、表征及生物活性研究[D].无锡:江南大学,2014.
[21]李雪芬,杜斌,丁轲,等.金属螯合肽分离纯化及其抗氧化活性的研究进展[J].中国食物与营养,2016,22(3):35-39.
[22]周亮.鱼鳞胶原小肽螯合锌的制备研究[D].武汉:湖北工业大学,2014.
[23]陈紫红,花朋朋,周文斌,等.多肽-矿质元素螯合物的研究进展[J].食品工业科技,2017,38(8):350-355.
[24]玄依凡.乳清蛋白肽—锌离子螯合物的制备及特性研究[D].哈尔滨:哈尔滨工业大学,2017.
[25] STORCKSDIECK S,BONSMANN G,HURRELL R F. Iron-binding properties, amino acid composition, and structure of muscle tissue peptides from in vitro digestion of different meat sources[J]. Journal of Food Science,2007,72(1):S019-S029.
[26] HUANG G,REN Z,JIANG J. Separation of iron-binding peptidesfrom shrimp processing by-products hydrolysates[J]. Food and Bioprocess Technology,2011,4(8):1 527-1 532.
[27] 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]. Journal of Agricultural and Food Chemistry,1998,46(1):49-53.
[28] PARK P J,JUNG W K,NAM K S,et al. Purification and characterization of antioxidative peptides from protein hydrolysate of lecithin-free egg yolk[J]. Journal of the American Oil Chemists'Society,2001,78(6):651-656.
[29] ZHANG L,LI J R,ZHOU K. Chelating and radical scavenging activities of soy protein hydrolysates prepared from microbial proteases and their effect on,eat lipid peroxidation[J]. Bioresource Technology,2010,101(7):2 084-2 089.
[30]左瑞雅. L-丙氨酰-L-谷氨酰胺-Zn2+螯合物制备及表征[D].重庆:重庆大学,2007.
[31]高素蕴.大豆多肽锌螯合盐的制备及生理活性研究[D].杨凌:西北农林科技大学,2003.
[32]张智,刘慧,刘奇,等.玉米肽-锌螯合物对小鼠免疫功能的影响[J].食品科技,2016,41(12):45-49.
[33]中本一雄.无机及配位化合物的红外和拉曼光谱(中译本)[M].北京:化学工业出版社,1981:257.
[34] LIAO W Z,LAI T,CHEN L Y,et al. Synthesis and characterization of a walnut peptides-zinc complex and its antiproliferative activity against human breast carcinoma cells through the induction of apoptosis[J]. Journal of Agricultural and Food Chemistry,2016,64(7):1 509-1 519.
[35]张智,刘慧,刘奇,等.玉米肽-锌螯合物结构表征及抗氧化活性分析[J].食品科学,2017,38(3):131-135.
[2]刘珊珊,敖静,谢宁宁,等.酪蛋白抗氧化肽的胃肠消化稳定性研究[J].中国食品学报,2014,14(2):47-54.
[3]高芸芳,王金水,苏银杰.酪蛋白结构及其酶解特性研究进展[J].农产品加工(学刊),2013(5):1-4; 9.
[4]于洋,祁艳霞,靳艳.乳源生物活性肽研究进展[J].食品与发酵工业,2017,43(9):259-266.
[5]张虎刚,张志翔,万端极.酪蛋白酶解制活性肽的工艺条件[J].食品研究与开发,2010,31(11):201-203.
[6]王战勇.牛乳酪蛋白ACE抑制肽的Caco-2细胞水平转运吸收机制研究[D].西安:陕西科技大学,2017.
[7]闫萍.大豆小分子多肽的抗疲劳作用及其机制研究[D].长春:吉林大学,2008
[8] PRASAD A S. Discovery of human zinc deficiency:50years later[J]. Journal of Trace Elements in Medicine and Biology,2012,26(2-3):66-69.
[9] XIE Ningning,HUANG Jingjing,CHENG Jianghua,et al.Affinity purification and characterization of zinc chelating peptides from rapeseed protein hydrolysates:Possible contribution of characteristic amino acid residues[J]. Food Chemistry,2015,173:210-217.
[10]高林,翟成杰,吴继红,等.改善人类锌营养途径的研究进展[J].食品安全质量检测学报,2013,4(5):1 550-1 555.
[11]曹继琼,何长华.锌缺乏对人体健康的影响[J].现代医药卫生,2014,30(7):1 016-1 019.
[12]陈丽丽,余江稳,潘博,等.克氏原螯虾壳肽螯合锌的制备其性质研究[J].江西科技师范大学学报,2017(6):65-70.
[13]林谢凤.鱼类副产物胶原肽螯合锌的制备及其性质研究[D].厦门:集美大学,2015.
[14]闫伟.真鲷鱼鳞皮胶原肽锌的制取、结构和性质研究[D].天津:天津科技大学,2013.
[15] HUSTINX J C A,SINGH T K,FOX P F. Heat-induced hydrolysis of sodium caseinate[J]. International Dairy Journal,1997,7(4):207-212.
[16] VAN BOEKEL M A J S. Heat-induced deamidation,dephosphorylation and breakdown of caseinate[J]. International Dairy Journal,1999,9(36):237-241.
[17]杨叶昆,李雪梅,唐自文,等.酪蛋白的酸水解及其棕色化反应产物在卷烟加香中的应用[J].烟草科技,2001(1):8-10.
[18]徐红梅,左凌波,高立娣.胰蛋白酶酶解酪蛋白工艺条件的优化[J].化工时刊,2015,29(3):1-5.
[19]荆莹.酪蛋白的酶水解改性研究[D].呼和浩特:内蒙古农业大学,2010.
[20]王晓萍.小麦胚芽蛋白源锌螯合肽的分离纯化、表征及生物活性研究[D].无锡:江南大学,2014.
[21]李雪芬,杜斌,丁轲,等.金属螯合肽分离纯化及其抗氧化活性的研究进展[J].中国食物与营养,2016,22(3):35-39.
[22]周亮.鱼鳞胶原小肽螯合锌的制备研究[D].武汉:湖北工业大学,2014.
[23]陈紫红,花朋朋,周文斌,等.多肽-矿质元素螯合物的研究进展[J].食品工业科技,2017,38(8):350-355.
[24]玄依凡.乳清蛋白肽—锌离子螯合物的制备及特性研究[D].哈尔滨:哈尔滨工业大学,2017.
[25] STORCKSDIECK S,BONSMANN G,HURRELL R F. Iron-binding properties, amino acid composition, and structure of muscle tissue peptides from in vitro digestion of different meat sources[J]. Journal of Food Science,2007,72(1):S019-S029.
[26] HUANG G,REN Z,JIANG J. Separation of iron-binding peptidesfrom shrimp processing by-products hydrolysates[J]. Food and Bioprocess Technology,2011,4(8):1 527-1 532.
[27] 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]. Journal of Agricultural and Food Chemistry,1998,46(1):49-53.
[28] PARK P J,JUNG W K,NAM K S,et al. Purification and characterization of antioxidative peptides from protein hydrolysate of lecithin-free egg yolk[J]. Journal of the American Oil Chemists'Society,2001,78(6):651-656.
[29] ZHANG L,LI J R,ZHOU K. Chelating and radical scavenging activities of soy protein hydrolysates prepared from microbial proteases and their effect on,eat lipid peroxidation[J]. Bioresource Technology,2010,101(7):2 084-2 089.
[30]左瑞雅. L-丙氨酰-L-谷氨酰胺-Zn2+螯合物制备及表征[D].重庆:重庆大学,2007.
[31]高素蕴.大豆多肽锌螯合盐的制备及生理活性研究[D].杨凌:西北农林科技大学,2003.
[32]张智,刘慧,刘奇,等.玉米肽-锌螯合物对小鼠免疫功能的影响[J].食品科技,2016,41(12):45-49.
[33]中本一雄.无机及配位化合物的红外和拉曼光谱(中译本)[M].北京:化学工业出版社,1981:257.
[34] LIAO W Z,LAI T,CHEN L Y,et al. Synthesis and characterization of a walnut peptides-zinc complex and its antiproliferative activity against human breast carcinoma cells through the induction of apoptosis[J]. Journal of Agricultural and Food Chemistry,2016,64(7):1 509-1 519.
[35]张智,刘慧,刘奇,等.玉米肽-锌螯合物结构表征及抗氧化活性分析[J].食品科学,2017,38(3):131-135.