响应面法制备小球藻多肽-钙螯合物的制备工艺
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摘要
为探究小球藻多肽-钙螯合物的较佳制备工艺,以小球藻干粉作为原料,对其进行酶解,获得小球藻多肽,以螯合率为指标,对小球藻多肽与CaCl_2的螯合工艺进行了优化,考查了螯合时间、螯合温度、肽钙质量比、底物质量分数和pH值对螯合反应的影响,通过Box-Behnken试验设计与响应面分析,同时建立螯合物制备工艺的二次项数学模型并验证其可靠性,优化后确定为螯合温度50℃,pH值9,肽钙质量比3∶1,底物质量分数5%,螯合时间60 min,最终螯合率可达96.14%。研究为合成新型补钙制剂(多肽-钙螯合物)的开发提供了理论依据,也为小球藻的开发利用提供了一个新的开拓思路。
The preparation of calcium-chelated Chlorella hydrolysis peptides was investigated in this study. The independent variables were pH,the solid-liquid ratio of Chlorella hydrolysis protein,addition levels of enzyme, reaction time,and temperature. The response was calcium chelating ability were systematically evaluated. The optimal parameters for chelating Chlorella hydrolysis peptides with CaCl_2 were obtained by using a 4-factor 3-level Box-Behnken experimental design with response surface methodology as follows:the solid-liquid ratio of Chelorella hydrolysis peptides was 5%,the hydrolysate and Ca Cl2 were mixed at 50 ℃ at a mass ratio of 3∶1 for 50 min and adjusted to pH 9. After reaction,the samples were collected by precipitating the calcium-chelated Chlorella hydrolysis peptides with pure ethanol,and dehydrated by vacuum freeze drying. The chelating rate was 96.14%. The results of this study can provide theoretical support and technical reference for the development of new calcium supplements and for the calcium-chelated Chlorella hydrolysis peptides product processing byproducts.
The preparation of calcium-chelated Chlorella hydrolysis peptides was investigated in this study. The independent variables were pH,the solid-liquid ratio of Chlorella hydrolysis protein,addition levels of enzyme, reaction time,and temperature. The response was calcium chelating ability were systematically evaluated. The optimal parameters for chelating Chlorella hydrolysis peptides with CaCl_2 were obtained by using a 4-factor 3-level Box-Behnken experimental design with response surface methodology as follows:the solid-liquid ratio of Chelorella hydrolysis peptides was 5%,the hydrolysate and Ca Cl2 were mixed at 50 ℃ at a mass ratio of 3∶1 for 50 min and adjusted to pH 9. After reaction,the samples were collected by precipitating the calcium-chelated Chlorella hydrolysis peptides with pure ethanol,and dehydrated by vacuum freeze drying. The chelating rate was 96.14%. The results of this study can provide theoretical support and technical reference for the development of new calcium supplements and for the calcium-chelated Chlorella hydrolysis peptides product processing byproducts.
引文
[1]杨璐生,李国平,陈林水.蛋白核小球藻粉的蛋白质、氨基酸含量及营养价值评价[J].亚热带植物科学,2003,32(1):36-38.
[2]刘世明.小球藻高密度异养培养[D].广州:华南理工大学,1999.
[3]时艳侠.小球藻藻种鉴定、株系筛选和培养条件优化[D].青岛:中国海洋大学,2007.
[4]唐旋,张豪,张倩,等.小球藻蛋白酶解肽铁配合物的制备及其性质研究[J].食品科技,2015(4):334-338.
[5]Sheih I C,Wu T K,Fang T J.Antioxidant properties of a new antioxidative peptide from algae protein waste hydrolysate in different oxidation systems[J].Bioresour Technol,2009(13):3 419-3 425.
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[7]甄守艳.羊骨胶原肽的酶法制备及肽钙螯合研究[D].晋中:山西农业大学,2015.
[8]黄顺丽,赵立娜,蔡茜茜,等.乳清蛋白钙螯合肽的分离及结构性质表征[J].中国食品学报,2015,15(11):212-218.
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[10]Jiang B,Mine Y.Phosphopeptides derived from hen egg yolk phosvitin:effects of molecular size on the calcium-binding properties[J].Biosci Biotechnol Biochem,2001(5):1 187-1 190.
[11]Bao X L,Song M,Zhang J,et al.Calcium-binding ability of soy protein hydrolysates[J].Chinese Chem Lett,2007(18):1 115-1 118.
[12]Chen D,Mu X,Huang H,et al.Isolation of a calciumbinding peptide from tilapia scale protein hydrolysateand its calcium bioavailability in rats[J].J Funct Foods,2014(6):575-584.
[13]Jung W K,Karawita R,Heo S J,et al.Recovery of a novel Ca-binding peptide from Alaska Pollack(Theragra chalcogramma)backbone by pepsinolytic hydrolysis[J].Process Biochem,2006,41(9):2 097-2 100.
[14]Reinhold J G,Lahimgarzadeh A,Nasr K,et al.Effects of purified phytate and phytate-rich bread upon metabolism of zinc,calcium,phosphorus,and nitrogen in man[J].Lancet,1973(7):283-288.
[15]Urbano G,Lopez-Jurado M,Fernandez M,et al.Ca and bioavailability of processed lentils as affected by dietary fiber and phytic acid content[J].Nutr Res,1999(19):49-54.
[16]Christakos S,Dhawan P,Porta A,et al.VD and intestinal calcium absorption[J].Mol Cell Endocrinol,2011(2):25-29.
[17]Perego S,Del F E,De L P,et al.Calcium bioaccessibility and uptake by human intestinal like cells following in vitro digestion of casein phosphopeptide-calcium aggregates[J].Food&Function,2015(6):1 796-1 807.
[18]张经坤,张泽民,于敖.人体钙吸收理论探讨[J].科学通报,2000,45(10):1 114-1 120.
[19]孟昌伟.斑点叉尾鮰鱼骨钙制剂的制备及其生物活性研究[D].合肥:合肥工业大学,2012.
[20]赵立娜,陈紫红,陈濠,等.响应面法优化菌草灵芝多肽-硒螯合物的制备工艺[J].食品科学,2017,38(14):187-192.
[21]赵立娜,汪少芸,张亚娇,等.响应面法优化乳清蛋白肽螯合钙离子的研究[J].中国食品学报,2014,14(4):64-71.
[22]马利华,秦卫东,陈学红,等.金针菇多糖-Fe(Ⅱ)螯合物的制备及抗氧化活性[J].食品科学,2010,31(20):202-207.
[23]金文刚,吴海,朱蓓薇,等.响应面优化虾夷扇贝生殖腺多肽-Ca2+螯合物制备工艺[J].食品科学,2013,34(16):11-16.
[24]丁利君,危雪如.罗非鱼蛋白酶解液的多肽与钙复合物的制备及其抑菌分析[J].食品科学,2009,30(20):198-202.
[25]张晓霞,刘盛取,李国英.碱法水解黑鱼鱼鳞及制备多肽螯合钙工艺的研究[J].食品科技,2010,35(12):130-134.
[26]毛学英,李明,李敏,等.不同因素对酪蛋白酶解产物螯合亚铁盐能力的影响[J].食品科学,2008,29(5):145-148.
[27]刘闪,刘良忠,李小娜,等.白鲢鱼骨胶原多肽螯合钙的工艺优化[J].食品科学,2014,35(10):76-81.
[28]鸿冰,汪之颖,刘鹏,等.鲢鱼骨胶原多肽螯合钙的制备研究[J].南方水产科学,2014,10(2):72-79.
[29]孙莉洁,梁金钟.响应面法优化大豆肽与钙离子螯合的研究[J].中国粮油学报,2010,25(1):22-27.
[30]张强,王松华,孙玉军,等.米糠蛋白酶解物锌螯合物的制备工艺研究[J].食品工业科技,2011,32(6):281-283.
[31]李靖.响应面优化灵芝多糖提取工艺研究[J].食品工业,2013(7):22-24.
[2]刘世明.小球藻高密度异养培养[D].广州:华南理工大学,1999.
[3]时艳侠.小球藻藻种鉴定、株系筛选和培养条件优化[D].青岛:中国海洋大学,2007.
[4]唐旋,张豪,张倩,等.小球藻蛋白酶解肽铁配合物的制备及其性质研究[J].食品科技,2015(4):334-338.
[5]Sheih I C,Wu T K,Fang T J.Antioxidant properties of a new antioxidative peptide from algae protein waste hydrolysate in different oxidation systems[J].Bioresour Technol,2009(13):3 419-3 425.
[6]梁春辉,菅景颖,张志胜.胶原多肽螯合钙壮骨作用研究[J].河北农业大学学报,2010,33(5):94-97.
[7]甄守艳.羊骨胶原肽的酶法制备及肽钙螯合研究[D].晋中:山西农业大学,2015.
[8]黄顺丽,赵立娜,蔡茜茜,等.乳清蛋白钙螯合肽的分离及结构性质表征[J].中国食品学报,2015,15(11):212-218.
[9]Holt C,Sawyer L.Primary and predicted secondary structures of the caseins in relation to their biological functions[J].Protein Eng,1988(2):251-259.
[10]Jiang B,Mine Y.Phosphopeptides derived from hen egg yolk phosvitin:effects of molecular size on the calcium-binding properties[J].Biosci Biotechnol Biochem,2001(5):1 187-1 190.
[11]Bao X L,Song M,Zhang J,et al.Calcium-binding ability of soy protein hydrolysates[J].Chinese Chem Lett,2007(18):1 115-1 118.
[12]Chen D,Mu X,Huang H,et al.Isolation of a calciumbinding peptide from tilapia scale protein hydrolysateand its calcium bioavailability in rats[J].J Funct Foods,2014(6):575-584.
[13]Jung W K,Karawita R,Heo S J,et al.Recovery of a novel Ca-binding peptide from Alaska Pollack(Theragra chalcogramma)backbone by pepsinolytic hydrolysis[J].Process Biochem,2006,41(9):2 097-2 100.
[14]Reinhold J G,Lahimgarzadeh A,Nasr K,et al.Effects of purified phytate and phytate-rich bread upon metabolism of zinc,calcium,phosphorus,and nitrogen in man[J].Lancet,1973(7):283-288.
[15]Urbano G,Lopez-Jurado M,Fernandez M,et al.Ca and bioavailability of processed lentils as affected by dietary fiber and phytic acid content[J].Nutr Res,1999(19):49-54.
[16]Christakos S,Dhawan P,Porta A,et al.VD and intestinal calcium absorption[J].Mol Cell Endocrinol,2011(2):25-29.
[17]Perego S,Del F E,De L P,et al.Calcium bioaccessibility and uptake by human intestinal like cells following in vitro digestion of casein phosphopeptide-calcium aggregates[J].Food&Function,2015(6):1 796-1 807.
[18]张经坤,张泽民,于敖.人体钙吸收理论探讨[J].科学通报,2000,45(10):1 114-1 120.
[19]孟昌伟.斑点叉尾鮰鱼骨钙制剂的制备及其生物活性研究[D].合肥:合肥工业大学,2012.
[20]赵立娜,陈紫红,陈濠,等.响应面法优化菌草灵芝多肽-硒螯合物的制备工艺[J].食品科学,2017,38(14):187-192.
[21]赵立娜,汪少芸,张亚娇,等.响应面法优化乳清蛋白肽螯合钙离子的研究[J].中国食品学报,2014,14(4):64-71.
[22]马利华,秦卫东,陈学红,等.金针菇多糖-Fe(Ⅱ)螯合物的制备及抗氧化活性[J].食品科学,2010,31(20):202-207.
[23]金文刚,吴海,朱蓓薇,等.响应面优化虾夷扇贝生殖腺多肽-Ca2+螯合物制备工艺[J].食品科学,2013,34(16):11-16.
[24]丁利君,危雪如.罗非鱼蛋白酶解液的多肽与钙复合物的制备及其抑菌分析[J].食品科学,2009,30(20):198-202.
[25]张晓霞,刘盛取,李国英.碱法水解黑鱼鱼鳞及制备多肽螯合钙工艺的研究[J].食品科技,2010,35(12):130-134.
[26]毛学英,李明,李敏,等.不同因素对酪蛋白酶解产物螯合亚铁盐能力的影响[J].食品科学,2008,29(5):145-148.
[27]刘闪,刘良忠,李小娜,等.白鲢鱼骨胶原多肽螯合钙的工艺优化[J].食品科学,2014,35(10):76-81.
[28]鸿冰,汪之颖,刘鹏,等.鲢鱼骨胶原多肽螯合钙的制备研究[J].南方水产科学,2014,10(2):72-79.
[29]孙莉洁,梁金钟.响应面法优化大豆肽与钙离子螯合的研究[J].中国粮油学报,2010,25(1):22-27.
[30]张强,王松华,孙玉军,等.米糠蛋白酶解物锌螯合物的制备工艺研究[J].食品工业科技,2011,32(6):281-283.
[31]李靖.响应面优化灵芝多糖提取工艺研究[J].食品工业,2013(7):22-24.