CN109122906A - 一种大米蛋白乳液及其制备方法 - Google Patents
一种大米蛋白乳液及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种大米蛋白乳液及其制备方法，所述方法包括：将大米蛋白溶于蒸馏水中并调节蛋白溶液的pH值及温度，然后蛋白溶液经过胰酶水解、离心、冷冻干燥处理得到大米蛋白水解物；将大米蛋白水解物溶于缓冲液，得到大米蛋白水解物溶液，将多酚加入大米蛋白水解物溶液中混合搅拌处理得到混合液，向混合液中加入大豆油后进行均质处理制得复合乳液。本方法所得复合物乳液工艺简单、操作安全，大米蛋白经酶解处理后与多酚复合，制得乳液具有较高的稳定性和抗氧化性、低过敏性。
Description
技术领域
本发明公开了一种大米蛋白乳液及其制备方法，属于食品加工技术领域。
背景技术
随着食品工业的迅速发展，人们越来越热衷于研究植物蛋白乳化性，并用其来替代传统乳化剂吐温等。大米蛋白作为我国人民膳食中的主要蛋白来源，具有其他植物蛋白不具备的低过敏性，并且其全面的氨基酸配比、降低胆固醇及高营养价值等特点使得大米蛋白逐渐成为研究热点。但是天然的大米蛋白由于其低溶解性，通常不能满足人们生产加工以及使用的要求。因此为了拓宽大米蛋白的应用范围，我们需要使大米蛋白改性从而提高其溶解性，最终改善其乳化性。目前提高蛋白溶解性的改性手段有脱酰胺、磷酸化、糖基化、酰基化等化学改性、以及加热、冷冻、挤压、微波、脉冲电场等物理改性。然而由于化学改性造成食品的毒性、过敏以及物理改性在较难实现产业化等，因此应用不广泛。目前酶法改性作为提高蛋白功能特性的主要方式，其具有反应条件温和，副产物少，专一性等优点，而且通过调节反应条件可以控制蛋白质的水解度，甚至可以有目的地选择酶切位点。
由表面活性剂制备的传统乳液体系界面能较大，液滴会很容易发生聚集并最终形成破乳现象，从而影响食品品质。新型的Pickering乳液是由固体颗粒稳定的乳液，目前在材料化工领域中研究比较多，固体颗粒常使用二氧化硅、石蜡球等。但无论是传统的表面的活性剂以及通常使用的固体颗粒稳定剂都具有一定毒性或者负面影响，都是不可食性，或者对人体具有一定的危害。目前食品领域研究较多的玉米醇溶蛋白、小麦醇溶蛋白等利用蛋白的双亲性来稳定乳液，但是此类蛋白只能作为稳定剂来提高乳液的贮藏期间的稳定性。乳液在贮藏期间会由于脂质的氧化，也是需要我们解决的重要问题，脂质的氧化会导致其乳液产品质量以及营养成分的损失。市场上常使用的乳液稳定剂并不能提供较高的氧化稳定性，需要控制贮藏条件、设计液滴界面等。
发明内容
本发明的目的在于提供一种大米蛋白乳液及其制备方法，达到增强大米蛋白利用性，提高乳液稳定性及抗氧化性的目的。
一种大米蛋白乳液的制备方法，包括如下步骤：
(1)将大米蛋白溶解于蒸馏水中得到质量分数为5%~10%的蛋白溶液，用0.5mol/L~1mol/L NaOH溶液调节蛋白溶液pH值到8.0~9.0，在45~55℃下加入大米蛋白含量0.5%~1%的酶进行酶解，酶解60~90min后在90~95℃灭酶处理10~ 15 min，经过离心、冻干得到大米蛋白水解物；
(2)将步骤(1)所得大米蛋白水解物溶于缓冲液制得质量分数为1%~5%的蛋白水解物溶液，搅拌30min~60min至完全溶解；
(3)将多酚类化合物加入步骤(2)所得溶液中，混合搅拌；然后向混合液中加入大豆油，所述的混合液与大豆油的体积比为4：1；
(4)将步骤(3)所得混合液进行高压均质处理得到高稳定性的复合乳液。
步骤2中所述缓冲液为10mM、pH值为7.0~9.0磷酸缓冲液。
步骤3所述多酚类化合物为绿原酸、没食子儿茶素没食子酸酯或没食子儿茶素。所述多酚类化合物的添加量为蛋白水解物质量的5%~10%。所述搅拌时间为6~12h。
步骤4所述高压均质为50~80MPa，时间为90~120 s。本发明还公开了采用上述方法制备得到的大米蛋白乳液。
本发明的有益效果：
（1）本发明采用酶解方式将蛋白结构打开，疏水性基团暴露，提高了未经改性的大米蛋白溶解性，利用蛋白质水解物与多酚的相互作用提高了大米蛋白的乳化，为解决传统的乳清蛋白、酪蛋白酸钠等乳化剂过敏源问题提供了一种新思路。
（2）本发明所制备的大米蛋白乳液在贮藏一个月之后仍然具有较好的分散性，前期实验表明，由2%~5%乳清蛋白制备的新鲜乳液粒径为289 nm~380 nm，经过一个月贮藏之后，乳液发生明显分层，粒径达到1.54 μm~2.24 μm，乳液分层极易影响产品最终的品质，大米蛋白乳液较好的解决了传统乳液稳定性问题，提高了植物源蛋白乳化剂在食品工业中应用的可能。
（3）本发明所制备的大米蛋白乳液具有较高的抗氧化性，在贮藏一个月期间，具有较好的抑制乳液氧化的能力，前期实验已证明，由2%~5%乳清蛋白制备的新鲜乳液POV值和TBARS值分别为5~10 meq/kg油，0.5~1.0 mmol/kg油，经过一个月贮藏之后变为了40~60meq/kg油、5~6 mmol/kg油，无法抑制乳液脂肪代谢，本发明制备的大米蛋白乳液中的多酚可以乳液氧化以及对食品乳液在贮藏过程中营养保存具有重要指导意义。
（3）本发明的制备工艺简单，反应条件温度，过程容易控制，生产周期短，设备投资和生产成本低，在食品行业具有广阔的应用前景。
（4）本发明的制备方法不添加有毒有害试剂，绿色安全、对环境不造成任何污染等特点。
具体实施方式
以下结合具体实施例对本发明作进一步说明。
实施例1：
将大米蛋白溶解于蒸馏水中得到质量分数为8%的蛋白溶液，用0.6mol/L NaOH溶液调节蛋白溶液pH到8.0，在50℃下加入大米蛋白含量0.5%的胰酶进行酶解，酶解60min后在90℃灭酶处理10min，经过离心、冻干得到大米蛋白水解物；将大米蛋白水解物溶于10mM、pH为8.6的磷酸缓冲液中，制得质量分数为1%的蛋白水解物溶液，搅拌30min至完全溶解；向上述溶液中加入大米蛋白水解物质量的6%的绿原酸，混合搅拌8 h；然后向混合液中加入大豆油，其中混合液与大豆油的体积比为4：1；将加入大豆油的混合液利用高压均质在70MPa处理 100 s得到高稳定性的大米蛋白乳液。
实施例2：
将大米蛋白溶解于蒸馏水中得到质量分数为10%的蛋白溶液，用1 mol/L NaOH 溶液调节蛋白溶液pH到9.0，在55℃下加入大米蛋白含量1%的胰酶进行酶解，酶解70 min后在93℃灭酶处理15 min，经过离心、冷冻干燥得到大米蛋白水解物；将大米蛋白水解物溶于10mM、pH为9.0的磷酸缓冲液中，制得质量分数为4%的蛋白水解物溶液，搅拌60min至完全溶解；向上述溶液中加入大米蛋白水解物质量的10%的绿原酸，混合搅拌6 h；然后向混合液中加入大豆油，其中混合液与大豆油的体积比为4：1；将加入大豆油的混合液利用高压均质在80MPa处理110 s得到高稳定性的大米蛋白乳液。
实施例3：
将大米蛋白溶解于蒸馏水中得到质量分数为5%的蛋白溶液，用0.8mol/L NaOH 溶液调节蛋白溶液pH到8.3，在48℃下加入大米蛋白含量0.8%的胰酶进行酶解，酶解80min后在95℃灭酶处理12min，经过离心、冷冻干燥得到大米蛋白水解物；将大米蛋白水解物溶于10mM、pH为8.2的磷酸缓冲液中，制得质量分数为3%的蛋白水解物溶液，搅拌50min至完全溶解；向上述溶液中加入大米蛋白水解物质量的5%的没食子儿茶素没食子酸酯，混合搅拌12 h；然后向混合液中加入大豆油，混合液与大豆油的体积比为4：1；将加入大豆油的混合液利用高压均质在70MPa处理90 s得到高稳定性的大米蛋白乳液。
实施例4：
将大米蛋白溶解于蒸馏水中得到质量分数为9%的蛋白溶液，用0.7 mol/L NaOH 溶液调节蛋白溶液pH到8.5，在45℃下加入大米蛋白含量0.9%的胰酶进行酶解，酶解80 min后在94°C灭酶处理14 min，经过离心、冷冻干燥得到大米蛋白水解物；将大米蛋白水解物溶于10mM、pH为7.8的磷酸缓冲液中，制得质量分数为2%的蛋白水解物溶液，搅拌50min至完全溶解；向上述溶液中加入大米蛋白水解物质量的6%的没食子儿茶素没食子酸酯，混合搅拌10h；然后向混合液中加入大豆油，混合液与大豆油的体积比为4：1；将加入大豆油的混合液利用高压均质在50 MPa处理120 s得到高稳定性的大米蛋白乳液。
实施例5：
将大米蛋白溶解于蒸馏水中得到质量分数为7%的蛋白溶液，用0.5 mol/L NaOH 溶液调节蛋白溶液pH到8.5，在54℃下加入大米蛋白含量0.6%的胰酶进行酶解，酶解90 min后在92℃灭酶处理10min，经过离心、冷冻干燥得到大米蛋白水解物；将大米蛋白水解物溶于10mM、pH为7.0的磷酸缓冲液中，制得质量分数为5%的蛋白水解物溶液，搅拌40min至完全溶解；向上述溶液中加入大米蛋白水解物质量的8%的没食子儿茶素，混合搅拌9 h；然后向混合液中加入大豆油，混合液与大豆油的体积比为4：1；将加入大豆油的混合液利用高压均质在60MPa处理110 s得到高稳定性的大米蛋白乳液。
大米蛋白乳液稳定性的考察：
粒径测定：将实施例1-5制备得到的乳液用去离子水稀释10000倍，采用MalvernMastersizer纳米粒度仪器测定乳液粒径。
POV值的测定：0.5 mL乳状液和10mL乙酸/氯仿溶液 (3:2，v/v)混合，加入0.5mL饱和碘化钾溶液，振荡半分钟，在暗处反应3min，加入30mL超纯水终止反应。加入0.5mL0.5%淀粉溶液，用2mmol/LNa2S2O3滴定至蓝色消失。用超纯水代替乳状液样品做空白。
POV(meq/kg油)=(S-B)×C/m
其中S和B分别是样品和空白所消耗的Na2S2O3的体积(mL)；C是Na2S2O3的浓度(mol/L)；m是样品的质量(g)。
TBARS值的测定：硫代巴比妥酸溶液的配制：0.375g硫代巴比妥酸，15g三氯乙酸，1.76mL、12mol/LHCl和82.9mL水混合加入3mL、2%BHA溶液（溶于乙醇），混匀。取2mL上述溶液与0.5mL样品及0.5mL水混匀，在沸水浴中加热15min，取出冷却到室温，5000r/min离心25min，取上清液在532nm处比色。以1,1,3,3-四乙氧基丙烷做标准曲线，计算样品TBARS值。测定结果如下表1所示：
表1大米蛋白乳液稳定性的研究
Claims (7)
1.一种大米蛋白乳液的制备方法，其特征在于，包括如下步骤：
(1)将大米蛋白溶解于蒸馏水中得到质量分数为5%~10%的蛋白溶液，用0.5mol/L~1mol/L NaOH溶液调节蛋白溶液pH值到8.0~9.0，在45~55℃下加入大米蛋白含量0.5%~1%的胰酶进行酶解，酶解60~90min后在90~95℃灭酶处理10~ 15min，经过离心、冻干得到大米蛋白水解物；
(2)将步骤(1)所得大米蛋白水解物溶于缓冲液制得质量分数为1%~5%的蛋白水解物溶液，搅拌30min~60min至完全溶解；
(3)将多酚类化合物加入步骤(2)所得溶液中，混合搅拌；然后向混合液中加入大豆油，所述的混合液与大豆油的体积比为4：1；
(4)将步骤(3)所得混合液进行高压均质处理得到高稳定性的复合乳液。
2.根据权利要求1所述的大米蛋白乳液的制备方法，其特征在于：所述步骤2中缓冲液为10mM、pH值为7.0~9.0的磷酸缓冲液。
3.根据权利要求1所述的大米蛋白乳液的制备方法，其特征在于：所述步骤3中多酚类化合物为绿原酸、没食子儿茶素没食子酸酯或没食子儿茶素。
4.根据权利要求1所述的大米蛋白乳液的制备方法，其特征在于：所述步骤3中多酚类化合物的添加量为大米蛋白水解物质量的5%~10%。
5.根据权利要求1所述的大米蛋白乳液的制备方法，其特征在于：所述步骤3中与多酚类化合物的搅拌时间为6~12h。
6.根据权利要求1所述的大米蛋白乳液的制备方法，其特征在于：所述步骤4中高压均质为50~80MPa，时间为90~120s。
7.采用权利要求1-6任一项所述方法制备得到的大米蛋白乳液。
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