酶解法制备豆粕水解肽工艺优化
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摘要
为制备苦味低且生物活性高的豆粕水解肽,本试验在综合考虑水解酶酶切位点后选取7种酶组合对低温豆粕进行水解,以蛋白质转化率、水解度、分子量分布及智能感官评价等作为水解肽的评价指标,比较不同酶组合的作用效果。结果表明,优化得到豆粕水解肽的最佳酶法制备工艺条件:复合酶的最佳组合为碱性蛋白酶+中性蛋白酶+脱苦蛋白酶,酶解pH值依次为8. 5、7. 5、7. 5,反应温度保持55℃,反应时间分别为3、2、2 h,经最佳酶组合制得的水解肽水解度为16%,分子量≤1 000 Da的水解肽可达89. 8%,制备的水解肽理化性质及口感更优。本研究结果为豆粕资源开发及功能性肽产品的应用奠定了理论基础。
In order to prepare hydrolysate peptides from soybean meal with low bitterness and high biological activity,seven different enzyme combinations were selected to prepare the hydrolyzed peptides with comprehensively considering their enzymatic cleavage sites. Protein conversion rate,degree of hydrolysis,molecular weight distribution and intelligent sensory evaluation were meausred as the evaluation indexs of the hydrolyzed peptides. The result showed that the optimum combination was alkaline protease + neutral protease + debittering enzymolysis,the pH value of enzymolysis solution was 8. 5,7. 5,and 7. 5,and the reaction time was 3 h,2 h,and 2 h,and the reaction temperature was kept at 55℃. After hydrolysis,the products showed degree of hydrolysis of 16%,and the distribution range of small-peptide molecular weight( ≤ 1 000 Da) was up to 89. 8%. The hydrolyzed peptides had better physical properties and textures.The results of this study provide a good foundation for the utilization of soybean protein and further application of functional peptides.
In order to prepare hydrolysate peptides from soybean meal with low bitterness and high biological activity,seven different enzyme combinations were selected to prepare the hydrolyzed peptides with comprehensively considering their enzymatic cleavage sites. Protein conversion rate,degree of hydrolysis,molecular weight distribution and intelligent sensory evaluation were meausred as the evaluation indexs of the hydrolyzed peptides. The result showed that the optimum combination was alkaline protease + neutral protease + debittering enzymolysis,the pH value of enzymolysis solution was 8. 5,7. 5,and 7. 5,and the reaction time was 3 h,2 h,and 2 h,and the reaction temperature was kept at 55℃. After hydrolysis,the products showed degree of hydrolysis of 16%,and the distribution range of small-peptide molecular weight( ≤ 1 000 Da) was up to 89. 8%. The hydrolyzed peptides had better physical properties and textures.The results of this study provide a good foundation for the utilization of soybean protein and further application of functional peptides.
引文
[1]杨小佳,王金水,管军军,杨国浩.豆粕的营养价值及影响因素[J].粮食与饲料工业,2013(3):44-46
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[3]王文娟,潘海涛,于磊娟.豆粕发酵制备大豆肽的研究[J].粮食加工,2007,32(2):55-56
[4]单春乔,吴磊,刘秋晨,刘艳,余丽芸.大豆小肽促进益生菌增殖作用的研究[J].中国微生态学杂志,2012,24(4):311-313
[5]Li J,Wang S Y,Zhang S M,Wang Y,Li H Z,Lin S.Preparation of angiotensin-Ⅰ-converting enzyme inhibitory peptides by multienzymatic hydrolysis of soybean meal[J].Food Science,2012,33(9):163-166
[6]Wen X Q,Liu S,Huang T,Zhao J X,Feng X,Sun J H.ACEinhibitory activity on hydrolytic peptides from soybean meal hydrolyzed by alkaline protease[J].Food Research&Development,2014,35(9):9-12
[7]Galvez A F.Products and methods using soy peptides to lower total and LDL cholesterol levels:US,US8598111[P].2018-03-08
[8]Yang R W,Wang J,Lin S Y.Isolation and purification of soybean antioxidant peptides[J].Advanced Materials Research,2014,881/883:811-814
[9]Xu J,Zhao Q,Qu Y,Ye F.Free radical scavenging activity of peptide fractions from defatted soybean meal hydrolysates evaluated by electron spin resonance[J].Food Science&Technology International,2013,19(6):557-566
[10]单春乔,吴磊,修立颖,文程,孙景秀,余丽芸.复合蛋白酶水解法制备大豆小肽的生产工艺研究[J].黑龙江畜牧兽医,2012,10(19):79-82
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[12]郝许峰.无机酸水解豆粕制备酱油的工艺研究[J].河南化工,2016,33(7):26-28
[13]吴飞林,赵明治,张瑶,熊智,徐平.蛋白质水解酶和样品预处理优化技术在蛋白质组学研究中的应用[J].生物工程学报,2016,32(3):306-316
[14]段志强.大豆肽的制备及其在食品中的应用[D].上海:华东师范大学,2015
[15]Wang S G,Yu S,Meng F G,Li B R,Song X G,Liu G F,Wang GL,Dai L,Gao P.Study on relative molecular weight distribution and depressor effect of soybean peptide prepared by enzymatic method[J].Science&Technology of Food Industry,2018,39(1):46-51
[16]Wang Z,Cui Y,Liu P,Zhao Y,Wang L,Liu Y,Xie J.Small peptides isolated from enzymatic hydrolyzate of fermented soybean meal promote endothelium-independent vasorelaxation and ACEinhibition[J].Journal of Agricultural&Food Chemistry,2017,65(50):10844-10850
[17]Chi C H,Cho S J.Improvement of bioactivity of soybean meal by solid-state fermentation with Bacillus amyloliquefaciens versus Lactobacillus spp.and Saccharomyces cerevisiae[J].LWT-Food Science and Technology,2016,68:619-625
[18]Adler N J.Control of the proteolytic reaction and of the level of bitterness in protein hydrolysis processes[J].Journal of Chemical Technology&Biotechnology,1984,34(3):215-222
[19]Xie Y,Liang X,Wei M,Zhao W,He B,Lu Q,Huo Q,Ma C.Optimization of glutamine peptide production from soybean meal and analysis of molecular weight distribution of hydrolysates[J].International Journal of Molecular Sciences,2012,13(6):7483-7495
[20]王慧云,陈海华,王雨生.复合酶双向酶解豆粕蛋白制备呈味肽研究[J].粮食与油脂,2014(5):26-28
[21]林洋,刘再胜,汲全柱,单春乔,倪天赫,刘秋晨,王凤忠,江国托.酶解法制备大豆小肽的工艺研究[J].大豆科学,2016,35(5):824-829
[22]高梅娟,刘平,兰小红,夏书芹,贾承胜,冯骉,张晓鸣.双酶酶解豆粕蛋白制备低苦味肽[J].食品工业科技,2010,31(2):193-197
[23]牟明峰,高云,宫关,孟建彪,张鹏.双酶水解制备大豆玉米活性多肽的研究[J].安徽农业科学,2012,40(7):3986-3988
[24]张晓峰,杨建平,潘春梅,卢亚兰.双酶法酶解豆粕蛋白最佳工艺条件研究[J].黑龙江畜牧兽医,2017(18):184-186
[25]李倩倩,罗旭,许腾,王艳,方芳,王凤忠.发芽对大豆中氨基酸含量的影响[J].核农学报,2018,32(3):548-554
[26]张群飞,徐大伦,杨文鸽.响应面法优化碱性蛋白酶酶解虾籽的工艺条件[J].核农学报,2014,28(5):876-882
[27]徐新娟,黄中文,王伟,贺亚飞.全自动凯氏定氮仪测定大豆蛋白质方法的研究[J].黑龙江农业科学,2016(2):108-110,121
[28]汪建斌,邓勇.Alcalase碱性蛋白酶对大豆分离蛋白水解作用的研究[J].食品工业科技,2002,23(1):61-63
[29]乙引,张显强,唐金刚,罗在柒.木瓜蛋白酶的纯化和性质[J].贵州师范大学学报(自然科学版),2002,20(1):11-14
[30]Bhattacharyya B K.Bromelain:An overview[J].Indian Journal of Natural Products and Resources,2008,7(4):359-363
[31]邓靖.脱苦蛋白酶霉菌菌种选育及酶学性质研究[D].长沙:湖南农业大学,2005
[32]谷海先,王建.风味蛋白酶的开发与利用[J].中国酿造,2003,22(1):26-28
[33]杨世平,邱德全.米曲霉中性蛋白酶特性的研究[J].广东海洋大学学报,2006,26(4):22-25
[34]肖凯军,高孔荣,曾庆孝.酶解大豆分离蛋白的特性研究[J].现代食品科技,1995,11(2):5-10
[35]鲁红.凝胶渗透色谱仪及其使用[J].分析仪器,2010(3):65-69
[36]关海宁,迟殿忠,刁小琴,王安然,迟程,冯立武,张甫.基于FAO/WHO相对氨基酸新评分模式的建立及其应用探讨[J].粮油加工,2008(12):114-116
[37]杨玉娟,姚怡莎,秦玉昌,邱静,李军国,李俊,谷旭.豆粕与发酵豆粕中主要抗营养因子调查分析[J].中国农业科学,2016,49(3):573-580
[38]张亚丽,徐忠.变性脱脂豆粕酶解物的特性研究[J].中国食品学报,2005,5(4):46-51
[39]黄雅燕,王文平,肖美添.碱性蛋白酶水解豆粕制备大豆多肽[J].华侨大学学报(自然科学版),2013,34(6):674-677
[40]李景,王素英,张思慜,王园,李昊泽,林菘.复合酶法水解豆粕制备ACE抑制肽[J].食品科学,2012,33(9):163-166
[41]邓靖,林亲录,赵谋明.酶法脱除蛋白水解产物苦味的研究进展[J].中国食品添加剂,2004(3):67-72
[42]Zhang Z G,Sun X Y.Study on enzymolysis and debittering process of soybean protein isolate by alkaline protease and circumscribed protease[J].Cereals&Oils,2016,29(8):53-57
[43]贾俊强,马海乐,王振斌,骆琳,何荣海.降血压肽的构效关系研究[J].中国粮油学报,2009,24(5):110-114
[2]曹钰,蔡国林,陆健.提高豆粕营养价值的研究进展[J].饲料与畜牧,2007(6):13-15
[3]王文娟,潘海涛,于磊娟.豆粕发酵制备大豆肽的研究[J].粮食加工,2007,32(2):55-56
[4]单春乔,吴磊,刘秋晨,刘艳,余丽芸.大豆小肽促进益生菌增殖作用的研究[J].中国微生态学杂志,2012,24(4):311-313
[5]Li J,Wang S Y,Zhang S M,Wang Y,Li H Z,Lin S.Preparation of angiotensin-Ⅰ-converting enzyme inhibitory peptides by multienzymatic hydrolysis of soybean meal[J].Food Science,2012,33(9):163-166
[6]Wen X Q,Liu S,Huang T,Zhao J X,Feng X,Sun J H.ACEinhibitory activity on hydrolytic peptides from soybean meal hydrolyzed by alkaline protease[J].Food Research&Development,2014,35(9):9-12
[7]Galvez A F.Products and methods using soy peptides to lower total and LDL cholesterol levels:US,US8598111[P].2018-03-08
[8]Yang R W,Wang J,Lin S Y.Isolation and purification of soybean antioxidant peptides[J].Advanced Materials Research,2014,881/883:811-814
[9]Xu J,Zhao Q,Qu Y,Ye F.Free radical scavenging activity of peptide fractions from defatted soybean meal hydrolysates evaluated by electron spin resonance[J].Food Science&Technology International,2013,19(6):557-566
[10]单春乔,吴磊,修立颖,文程,孙景秀,余丽芸.复合蛋白酶水解法制备大豆小肽的生产工艺研究[J].黑龙江畜牧兽医,2012,10(19):79-82
[11]San J S,Rai A K.Production of bioactive peptides during soybean fermentation and their potential health benefits[J].Trends in Food Science&Technology,2016,50:1-10
[12]郝许峰.无机酸水解豆粕制备酱油的工艺研究[J].河南化工,2016,33(7):26-28
[13]吴飞林,赵明治,张瑶,熊智,徐平.蛋白质水解酶和样品预处理优化技术在蛋白质组学研究中的应用[J].生物工程学报,2016,32(3):306-316
[14]段志强.大豆肽的制备及其在食品中的应用[D].上海:华东师范大学,2015
[15]Wang S G,Yu S,Meng F G,Li B R,Song X G,Liu G F,Wang GL,Dai L,Gao P.Study on relative molecular weight distribution and depressor effect of soybean peptide prepared by enzymatic method[J].Science&Technology of Food Industry,2018,39(1):46-51
[16]Wang Z,Cui Y,Liu P,Zhao Y,Wang L,Liu Y,Xie J.Small peptides isolated from enzymatic hydrolyzate of fermented soybean meal promote endothelium-independent vasorelaxation and ACEinhibition[J].Journal of Agricultural&Food Chemistry,2017,65(50):10844-10850
[17]Chi C H,Cho S J.Improvement of bioactivity of soybean meal by solid-state fermentation with Bacillus amyloliquefaciens versus Lactobacillus spp.and Saccharomyces cerevisiae[J].LWT-Food Science and Technology,2016,68:619-625
[18]Adler N J.Control of the proteolytic reaction and of the level of bitterness in protein hydrolysis processes[J].Journal of Chemical Technology&Biotechnology,1984,34(3):215-222
[19]Xie Y,Liang X,Wei M,Zhao W,He B,Lu Q,Huo Q,Ma C.Optimization of glutamine peptide production from soybean meal and analysis of molecular weight distribution of hydrolysates[J].International Journal of Molecular Sciences,2012,13(6):7483-7495
[20]王慧云,陈海华,王雨生.复合酶双向酶解豆粕蛋白制备呈味肽研究[J].粮食与油脂,2014(5):26-28
[21]林洋,刘再胜,汲全柱,单春乔,倪天赫,刘秋晨,王凤忠,江国托.酶解法制备大豆小肽的工艺研究[J].大豆科学,2016,35(5):824-829
[22]高梅娟,刘平,兰小红,夏书芹,贾承胜,冯骉,张晓鸣.双酶酶解豆粕蛋白制备低苦味肽[J].食品工业科技,2010,31(2):193-197
[23]牟明峰,高云,宫关,孟建彪,张鹏.双酶水解制备大豆玉米活性多肽的研究[J].安徽农业科学,2012,40(7):3986-3988
[24]张晓峰,杨建平,潘春梅,卢亚兰.双酶法酶解豆粕蛋白最佳工艺条件研究[J].黑龙江畜牧兽医,2017(18):184-186
[25]李倩倩,罗旭,许腾,王艳,方芳,王凤忠.发芽对大豆中氨基酸含量的影响[J].核农学报,2018,32(3):548-554
[26]张群飞,徐大伦,杨文鸽.响应面法优化碱性蛋白酶酶解虾籽的工艺条件[J].核农学报,2014,28(5):876-882
[27]徐新娟,黄中文,王伟,贺亚飞.全自动凯氏定氮仪测定大豆蛋白质方法的研究[J].黑龙江农业科学,2016(2):108-110,121
[28]汪建斌,邓勇.Alcalase碱性蛋白酶对大豆分离蛋白水解作用的研究[J].食品工业科技,2002,23(1):61-63
[29]乙引,张显强,唐金刚,罗在柒.木瓜蛋白酶的纯化和性质[J].贵州师范大学学报(自然科学版),2002,20(1):11-14
[30]Bhattacharyya B K.Bromelain:An overview[J].Indian Journal of Natural Products and Resources,2008,7(4):359-363
[31]邓靖.脱苦蛋白酶霉菌菌种选育及酶学性质研究[D].长沙:湖南农业大学,2005
[32]谷海先,王建.风味蛋白酶的开发与利用[J].中国酿造,2003,22(1):26-28
[33]杨世平,邱德全.米曲霉中性蛋白酶特性的研究[J].广东海洋大学学报,2006,26(4):22-25
[34]肖凯军,高孔荣,曾庆孝.酶解大豆分离蛋白的特性研究[J].现代食品科技,1995,11(2):5-10
[35]鲁红.凝胶渗透色谱仪及其使用[J].分析仪器,2010(3):65-69
[36]关海宁,迟殿忠,刁小琴,王安然,迟程,冯立武,张甫.基于FAO/WHO相对氨基酸新评分模式的建立及其应用探讨[J].粮油加工,2008(12):114-116
[37]杨玉娟,姚怡莎,秦玉昌,邱静,李军国,李俊,谷旭.豆粕与发酵豆粕中主要抗营养因子调查分析[J].中国农业科学,2016,49(3):573-580
[38]张亚丽,徐忠.变性脱脂豆粕酶解物的特性研究[J].中国食品学报,2005,5(4):46-51
[39]黄雅燕,王文平,肖美添.碱性蛋白酶水解豆粕制备大豆多肽[J].华侨大学学报(自然科学版),2013,34(6):674-677
[40]李景,王素英,张思慜,王园,李昊泽,林菘.复合酶法水解豆粕制备ACE抑制肽[J].食品科学,2012,33(9):163-166
[41]邓靖,林亲录,赵谋明.酶法脱除蛋白水解产物苦味的研究进展[J].中国食品添加剂,2004(3):67-72
[42]Zhang Z G,Sun X Y.Study on enzymolysis and debittering process of soybean protein isolate by alkaline protease and circumscribed protease[J].Cereals&Oils,2016,29(8):53-57
[43]贾俊强,马海乐,王振斌,骆琳,何荣海.降血压肽的构效关系研究[J].中国粮油学报,2009,24(5):110-114