CN113907353A - 一种提高姜黄素水溶性和小肠消化稳定性的方法 - Google Patents
一种提高姜黄素水溶性和小肠消化稳定性的方法 Download PDFInfo
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Abstract
本发明提供一种提高姜黄素水溶性和小肠消化稳定性方法，包括：a)从麦芽中分离纯化Protein Z，溶解于缓冲液中，得到2～10μM Protein Z溶液；b)将过量姜黄素加入到Protein Z溶液中，震荡，将得到的Protein Z和姜黄素的混合物过水系滤膜；c)将过滤得到的Protein Z‑姜黄素的水溶液和胰蛋白酶溶液以Protein Z/胰蛋白酶的摩尔比＝1:1～1:10混合加入到体外模拟小肠消化体系中。本发明以天然麦芽中的Protein Z为主要原料，将其应用于提高姜黄素的溶解度和消化稳定性。姜黄素与Protein Z发生相互作用，对姜黄素进行有效保护，进而提高姜黄素的生物利用度。
Description
技术领域
本发明属于姜黄素应用技术领域，尤其涉及一种提高姜黄素水溶性和小肠消化稳定性的方法。
背景技术
姜黄素是姜黄中含量最多的一种天然的色素，最古以来就被当成一种食品添加剂用于食品加工过程中。除此之外，姜黄素作为一种多酚类化合物，具有抗炎、抗肿瘤、抗氧化、抗菌以及改善肠胃功能等多种生物活性功效。然而，尽管姜黄素具备多种生物学功能，但它在水溶液中的溶解度极低，以及极易在生理条件下发生降解等问题使其难以有效应用于食品体系，并进一步限制了机体的吸收。Protein Z是啤酒中最早被发现拥有较高表面粘度和弹性的蛋白质，主要来源于大麦胚乳，分子量在43kDa左右，它能在啤酒酿造过程中抵抗酶类的降解、高温的影响，最终成为啤酒中含量最为丰富的蛋白质。前期研究发现，Protein Z可以与啤酒中多种苦味小分子发生相互作用从而提高啤酒的泡沫稳定性并进一步对小分子进行保护。因此，将Protein Z作为应用载体用于提高姜黄素的水溶性以及消化稳定性是可行的，目前尚未有关Protein Z用于提高小分子化合物溶解性以及稳定性的报道。此外，利用Protein Z维持泡沫的特性，将Protein Z-姜黄素应用于气泡饮料不仅可以赋予其丰富的气泡，还为开发具有抗氧化等功能性饮料提供思路。
发明内容
有鉴于此，本发明的目的在于提供一种提高姜黄素水溶性和小肠消化稳定性的方法，该方法制得的protein Z蛋白溶液用于提高姜黄素在水溶液中的溶解度，并且能有效提高姜黄素在体外模拟小肠消化体系的稳定性。
本发明提供了一种提高姜黄素水溶性和小肠消化稳定性的方法，其特征在于：所述的制备方法包括如下步骤：
a)从麦芽中分离纯化Protein Z，溶解于缓冲液中，制备浓度为2～10μMProtein Z溶液；
b)将过量姜黄素样品加入到2～10μM Protein Z溶液中，在摇床4～37℃中以100～220r/min震荡2～24h，将得到的Protein Z和姜黄素的混合物过0.45μm水系滤膜，过滤得到Protein Z-姜黄素的水溶液；
c)将Protein Z-姜黄素的水溶液和胰蛋白酶溶液以ProteinZ/胰蛋白酶的摩尔比＝1:1～1:10的比例混合加入到体外模拟小肠消化体系中。
本发明以天然麦芽中的Protein Z为主要原料，利用其高温稳定性、溶解性好、抑制蛋白酶活性等优点，将其应用于提高姜黄素的溶解度和消化稳定性。姜黄素可以与Protein Z发生相互作用，对姜黄素进行有效保护，进而提高姜黄素的生物利用度。
本发明借助Protein Z蛋白的水溶性好、热稳定性好、以及广泛存在于麦芽、啤酒等大麦制品中等优点，制备Protein Z-姜黄素复合物，利用蛋白质对小分子化合物的保护作用，提高水溶液中姜黄素的溶解度和消化稳定性。
优选地，所述缓冲液选自10mM Tris-HCl,pH值＝7.0～9.0；或10mM Mops,pH值＝6.5～7.9；或10mMPBS,pH值＝7.3。
优选地，所述Protein Z作为改善姜黄素溶解度的蛋白载体，提高姜黄素的水溶性以及消化稳定性。此外，由于Protein Z作为一种食源性材料可以添加至食品饮料中，能有效拓展姜黄素在食品工业中的应用前景，提高食品的感官特性。
优选地，所述Protein Z溶液有效提高姜黄素在水溶液中的溶解度，姜黄素在蛋白溶液中的溶解度提高了20倍以上。
优选地，所述Protein Z将姜黄素在体外模拟小肠消化体系中的稳定性提高60％以上。
优选地，Protein Z-姜黄素复合物具有起泡性和泡沫稳定性。
在本发明中，所述姜黄素是姜黄属植物块茎中主要的治疗剂，可用作膳食补充剂、着色剂以及化妆品。姜黄素具有抗氧化、抗炎、抗病毒和抗菌等多种生物学活性，然而极低的水溶性和较低的肠胃消化稳定性极大地限制了其生物利用度。
本发明提供了一种提高姜黄素水溶性和小肠消化稳定性的方法，包括如下步骤：a)从麦芽中分离纯化Protein Z，溶解于缓冲液中，制备浓度为2～10μMProtein Z溶液；b)将过量姜黄素样品加入到2～10μMProtein Z溶液中，在摇床4～37℃中以100～220r/min震荡2～24h，将得到的ProteinZ和姜黄素的混合物过0.45μm水系滤膜，过滤得到Protein Z-姜黄素的水溶液；c)将Protein Z-姜黄素的水溶液和胰蛋白酶溶液以Protein Z/胰蛋白酶＝1:1～1:10的比例混合加入到体外模拟小肠消化体系中。
与现有技术相比，本发明的有益效果在于：
本发明步骤简单，操作简便，无需复杂的仪器设备，成本相对较低，绿色清洁，对环境无污染。
本发明方法制备的提高姜黄素溶解度的蛋白溶液利用食源性原料作为应用的载体，可以有效提高姜黄素的溶解度和消化稳定性，可以极大的改善姜黄素在机体内的吸收、代谢效率从而进一步提高其生物利用率。
本发明可以应用于食品(如啤酒、功能饮料、气泡饮料等)、医药、化妆品等多个领域，具有良好的市场前景。
附图说明
图1为不同浓度的Protein Z中溶解的姜黄素样品；
图2为体外模拟小肠消化体系中消化不同时间(0、30、60、120min)后的姜黄素含量。
具体实施方式
为了进一步说明本发明，下面结合实施例对本发明提供的一种提高姜黄素水溶性和小肠消化稳定性的方法进行详细地描述，但不能将它们理解为对本发明保护范围的限定。
实施例1
将Protein Z蛋白样品溶于10mMMops(pH7.0)缓冲液中，分别配置成0、2、4、6、8、10μM浓度梯度的Protein Z蛋白样品。取1mL不同浓度的Protein Z样品，分别加入过量的姜黄素样品，避光混匀24h，过0.45μm滤膜，得到溶解有姜黄素的protein Z蛋白溶液。利用紫外分光光度计测定425nm处的吸光度值检测样品中溶解得到的姜黄素含量，检测得到姜黄素在水溶液的溶解度从0.43μM增加到10.59μM，提升了约24倍。
实施例2
将Protein Z蛋白溶液利用10mMMops(pH 7.0)缓冲液配置成212μM蛋白样品；将姜黄素溶于无水乙醇配置成5mM均一溶液；将胰蛋白酶溶于10mMMops(pH7.0)缓冲液配置成50μM蛋白酶样品。按混合体积5mL，Protein Z(8μM):姜黄素(n/n)＝1:3，protein Z:胰蛋白酶(n/n)＝1:4来混合上述溶液，利用10mM Mops(pH7.0)缓冲液补足5mL反应体系，将不添加Protein Z设置为对照组。将样品置于37℃反应，分别在0、30，60，120min取样，煮沸5min终止反应。取终止反应的样品450μL加450μL乙酸乙酯萃取2h，通过HPLC检测体系中未被降解的姜黄素含量，相比于对照组体系中的姜黄素含量为15.13μM，添加protein Z后体系中姜黄素含量为24.76μM，稳定性提高了约60％。
由以上实施例可知，本发明提供了一种提高姜黄素水溶性和小肠消化稳定性的方法，包括如下步骤：a)从麦芽中分离纯化Protein Z，溶解于缓冲液中，制备浓度为2～10μMProtein Z溶液；b)将过量姜黄素样品加入到2～10μMProtein Z溶液中，在摇床4～37℃中以100～220r/min震荡2～24h，将得到的Protein Z和姜黄素的混合物过0.45μm水系滤膜，过滤得到Protein Z-姜黄素的水溶液；c)将Protein Z-姜黄素的水溶液和胰蛋白酶溶液以Protein Z/胰蛋白酶＝1:1～1:10的比例混合加入到体外模拟小肠消化体系中。实验结果表明：添加protein Z后体系中姜黄素含量为24.76μM，稳定性提高了约60％；姜黄素在水溶液的溶解度从0.43μM增加到10.59μM，提升了约24倍。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (6)
1.一种提高姜黄素水溶性和小肠消化稳定性的方法，其特征在于：包括如下步骤：
a)从麦芽中分离纯化Protein Z，溶解于缓冲液中，制备浓度为2～10μM Protein Z溶液；
b)将过量姜黄素样品加入到2～10μM Protein Z溶液中，在摇床4～37℃中以100～220r/min震荡2～24h，将得到的Protein Z和姜黄素的混合物过0.45μm水系滤膜，过滤得到Protein Z-姜黄素的水溶液；
c)将Protein Z-姜黄素的水溶液和胰蛋白酶溶液以ProteinZ/胰蛋白酶的摩尔比＝1:1～1:10的比例混合加入到体外模拟小肠消化体系中。
2.根据权利要求1所述的方法，其特征在于，所述缓冲液选自10mM Tris-HCl,pH值＝7.0～9.0；或10mM Mops,pH值＝6.5～7.9；或10mMPBS,pH值＝7.3。
3.根据权利要求1所述的方法，其特征在于，所述Protein Z作为改善姜黄素溶解度的蛋白载体。
4.根据权利要求1所述的方法，其特征在于，所述Protein Z溶液有效提高姜黄素在水溶液中的溶解度，姜黄素在蛋白溶液中的溶解度提高了20倍以上。
5.根据权利要求1所述的方法，其特征在于，所述Protein Z将姜黄素在体外模拟小肠消化体系中的稳定性提高60％以上。
6.根据权利要求1所述的方法，其特征在于，Protein Z-姜黄素复合物具有起泡性和泡沫稳定性。
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