CN106901311B - 呈味肽和应用 - Google Patents
呈味肽和应用 Download PDFInfo
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- CN106901311B CN106901311B CN201510971791.7A CN201510971791A CN106901311B CN 106901311 B CN106901311 B CN 106901311B CN 201510971791 A CN201510971791 A CN 201510971791A CN 106901311 B CN106901311 B CN 106901311B
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- asn
- taste
- umami
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Images
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Abstract
本发明涉及呈味肽和应用。具体而言,本发明涉及一种三肽,所述三肽由选自Ser、Pro、Asn、Ala和Leu中的任意三种氨基酸组成。本发明的三肽优选能提供浓厚味、鲜味、咸味、甜味和/或酸味,更优选具有优异的酸味和鲜味味觉作用,且具有优异的稳定性。
Description
技术领域
本发明涉及呈味肽和应用。
背景技术
食品中存在许多呈味物质(例如甜味物质、鲜味物质、咸味物质、酸味物质等),这些呈味物质具有复杂的呈味功能,能同时参与并影响食品的滋味的形成,能提高食品的总体风味,改进食品的质构,使食品的总体味感协调、细腻、醇厚浓郁。
鲜味作为基本味道之一被描述为可口或肉汤的风味,是许多美味食物产品特有的味觉属性。目前食品工业普遍认可的鲜味物质为谷氨酸衍生物谷氨酸钠。已知通过添加谷氨酸盐,特别是谷氨酸钠(MSG)可以改变或增加食品的鲜味和美味感受,但近年来科学研究报道认为谷氨酸钠可能对人体带来一些负面影响,受此影响消费者也开始倾向于减少甚至不选择谷氨酸钠作为美味增加剂,以期减少对其依赖性。基于此,食品工业者寻找到可以替代谷氨酸钠的物质,也即通过添加天然存在的5’-肌苷酸盐和5’-鸟苷酸盐或其混合物增加食品产品的鲜味和美味特性。但这些物质同时存在有生产过程复杂、成本较高等缺陷,从而限制了在食品工业中的应用。
CN 104650073A公开了一种可用作鲜味剂的化合物,该化合物可单独用作鲜味剂或与谷氨酸钠、5’-肌苷酸盐和5’-鸟苷酸盐随意组合使用,但该鲜味剂来源为化学物质,其安全性、应用性仍待研究。与此同时,对食品本身中高含量的蛋白质、肽类和氨基酸的研究发现,食物中鲜味物质除谷氨酸、天冬氨酸等氨基酸外,存在于食物中的肽类同样可以提供鲜味和可口味觉的呈味特性。目前已发现的食物中存在的鲜味或美味肽主要有:源于面筋水解蛋白的pGlu-Pro-Ser、pGlu-Pro、pGlu-Pro-Glu和pGlu-Pro-Gln,源于花生水解蛋白的Ser-Ser-Arg-Asn-Glu-Gln-Ser-Arg和Glu-Gly-Ser-Glu-Ala-Pro-Asp-Gly-Ser-Ser-Arg以及源于河豚肉的Tyr-Gly-Gly-Glu-Thr-Pro-Pro-Phe-Val。这些鲜味多肽的分离鉴定说明可以通过合成鲜味呈味多肽替代食品工业应用中大量存在的鲜味化合物。
发明内容
本发明第一方面提供一种三肽,所述三肽由选自Ser、Pro、Asn、Ala和Leu中的任意三种氨基酸组成。
在某些实施方案中,所述三肽的一个氨基酸为Asn。
在某些实施方案中,所述Asn位于所述三肽的C端。
在某些实施方案中,所述三肽选自:
(1)Ser-Pro-Asn;和
(2)Ala-Leu-Asn。
在某些具体实施方案中,所述呈味肽提供鲜味。
在某些具体实施方案中,所述呈味肽提供酸味。
本发明第二方面提供一种组合物,所述组合物含有由选自Ser、Pro、Asn、Ala和Leu中的任意三种氨基酸组成的三肽。
在某些具体实施方式中,所述三肽的一个氨基酸为Asn。
在某些具体实施方式中,所述三肽选自:
(1)Ser-Pro-Asn;和
(2)Ala-Leu-Asn。
在某些具体实施方案中,所述组合物是调味品。
在某些具体实施方案中,所述调味品为酱料、酱油、奶酪涂抹料、汤料、味精。
在某些具体实施方案中,所述调味品还含有5’-肌苷酸盐、5’-鸟苷酸盐、琥珀酸及其盐、植物水解蛋白、酵母抽提物、N1-(2,4-二甲氧基苄基)-N2-(2-(2-吡啶基)乙基)草酰胺、鲜味氨基酸、鲜味呈味肽和厚味呈味肽(谷胱甘肽、谷缬甘肽等)中的一种或任意多种的混合物。
在某些具体实施方案中,所述组合物是食品。
在某些具体实施方案中,所述食品选自发酵食品、保健食品、传统面制品和烘焙食品。
在某些具体实施方案中,所述食品选自油脂产品、饮料、汤、冰激凌、酸奶、蛋糕调和粉、和奶酪。
在某些具体实施方案中,所述调味品是鲜味调味品。
在某些具体实施方案中,所述调味品是酸味调味品。
在某些具体实施方案中,所述组合物中,所述三肽的含量为0.01~20%,优选0.1~10%。
本发明还提供一种增加食物鲜味和/或酸味的方法,所述方法包括向所述食物中加入由选自Ser、Pro、Asn、Ala和Leu中的任意三种氨基酸组成的三肽,或含所述三肽的组合物。
在某些具体实施方式中,所述三肽的一个氨基酸为Asn。
在某些具体实施方式中,所述三肽选自:
(1)Ser-Pro-Asn;和
(2)Ala-Leu-Asn。
本发明还包括本发明三肽在增加食物或调味浓厚味、鲜味、咸味、甜味和/或酸味,尤其是品鲜味和/或酸味中的应用,或在制备用于增加食物或调味品浓厚味、鲜味、咸味、甜味和/或酸味,尤其是鲜味和/或酸味的调味品或食品中的应用。
附图说明
图1A和1B分别显示Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽HPLC检测纯度图谱。
图2A和2B分别显示Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽质谱鉴定氨基酸序列图谱。
图3显示市售调味汁复配Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽效果图。
具体实施方式
本发明基于食品工业对于可用于美味食品风味增强物质的需求,提供具有浓厚味、鲜味、咸味、甜味和酸味中的任一种或任意多种味道的呈味肽,尤其是具有酸味和/或鲜味的呈味肽。本发明的呈味肽可替代食品工业应用中大量存在的浓厚味、鲜味、咸味、甜味和/或酸味,尤其是酸味和/或鲜味化合物。
因此,本发明提供的呈味肽由选自Ser、Pro、Asn、Ala和Leu中的任意三种氨基酸组成。该呈味肽优选能提供浓厚味、鲜味、咸味、甜味和/或酸味,更优选具有优异的酸味和鲜味味觉作用,且具有优异的稳定性。因此,优选的是,本发明的呈味肽为酸味和/或鲜味呈味肽。
优选的是,本发明的呈味肽中至少包括Asn。更优选的是,本发明呈味肽中Asn位于C端。
作为示例性例子,本发明的三肽可具有以下结构:Ser-Pro-Asn、Ala-Leu-Asn、Leu-Ala-Asn,Pro-Ser-Asn,Asn-Ser-Pro,Asn-Pro-Ser,Ser-Asn-Pro,Pro-Asn-Ser,Ala-Asn-Leu,Leu-Asn-Ala,Asn-Ala-Leu,Asn-Leu-Ala。
可采用人工合成的方法合成本发明的呈味肽。
对于人工合成,可采用本领域周知的多肽化学合成法来合成本发明的氨基酸序列。多肽化学合成方法包括固相合成法和液相合成法,其中以固相合成法常用。固相合成方法包括但不限于Fmoc和tBoc两种常用方法。通常,使用树脂作为不溶性的固相载体,通常从C端(羧基端)向N端(氨基端)逐个将氨基酸连接在肽链上,每个氨基酸连接循环由以下三步反应构成:1)脱保护:被保护的氨基酸必须用一种脱保护溶剂去除氨基的保护基团;2)活化:待连接的氨基酸的羧基被活化剂所活化;和3)偶联:活化的羧基与前一个氨基酸裸露的氨基反应,形成肽键。反复循环直到肽链延伸至所需长度时即可完成。最后用切割液切割肽链和固相载体之间的连接,就可获得所需的氨基酸序列。
可以在程序控制的自动化多肽合成仪上进行上述化学合成,这类仪器包括但不限于Protein Technologies公司推出的Tribute双通道多肽合成仪、CS Bio公司的UV OnlineMonitor***、Aapptec公司推出的Focus XC三通道合成仪等。
合成本发明的三肽之后,可采用HPLC检测合成物纯度,UPLC-Q-TOF-MS/MS鉴定合成物结构,并采用分子感官科学方法评价呈味肽合成物味觉作用效果。
本发明的三肽也可从豆类、谷物类、杂粮类、微生物类等植物蛋白来源中通过发酵、酶解等方法制备,并通过离心、过滤、超滤、微滤、反渗透及体积排阻色谱、离子交换色谱、反相色谱、疏水作用色谱、亲和色谱等方法分离纯化获得。
本发明的呈味肽可作为基料或辅料,广泛使用在食品领域中,例如用于汁、酱、酱油、汤、发酵食品等调味呈味剂中。本发明的呈味肽还可以用于配制其他食用制品,包括但不限于油脂产品、调味料、汤料、保健食品、小吃、饮料、冰激凌、酸奶、蛋糕调和粉、传统面制品、烘焙食品(例如,面包和松饼),奶酪和奶酪涂抹料等。
同时,该呈味肽可复配其他调味物质如:盐、味精,5’-肌苷酸盐、5’-鸟苷酸盐、琥珀酸及其盐、植物水解蛋白、酵母抽提物、N1-(2,4-二甲氧基苄基)-N2-(2-(2-吡啶基)乙基)草酰胺、鲜味氨基酸、鲜味呈味肽、厚味呈味肽(谷胱甘肽、谷缬甘肽等)及其组合物等。
因此,本发明提供一种呈味肽的组合物。具体而言,该组合物含有本发明的呈味肽,具体为由选自Ser、Pro、Asn、Ala和Leu中的任意三种氨基酸组成的三肽。优选的是,本发明的组合物含有Ser-Pro-Asn、Ala-Leu-Asn或两者。
组合物中,呈味肽的含量通常为0.01~20%,优选0.1~10%,例如0.1~5%、0.1~2%不等。更优选的是,在调味品中,所述呈味肽的含量为0.01~20%,优选0.1~10%,例如0.1~5%、0.1~2%不等。
该组合物可作为调味品使用。调味品通常包括酱料、酱油、奶酪涂抹料、汤料和味精。调味品中还可含有5’-肌苷酸盐、5’-鸟苷酸盐、琥珀酸及其盐、植物水解蛋白、酵母抽提物、N1-(2,4-二甲氧基苄基)-N2-(2-(2-吡啶基)乙基)草酰胺、鲜味氨基酸、鲜味呈味肽和厚味呈味肽中的一种或任意多种的混合物。
本发明中,鲜味氨基酸可以是谷氨酸、天冬氨酸、丝氨酸、谷氨酰胺、天冬酰胺等。
鲜味呈味肽可以是Glu-Glu,Glu-Asp,Leu-Asp-Leu等。
厚味呈味肽可以是谷胱甘肽和谷缬甘肽。
组合物可以是食品,例如发酵食品、保健食品、传统面制品和烘焙食品。或者,食品可以是油脂产品、饮料、汤、冰激凌、酸奶、蛋糕调和粉、和奶酪。
换言之,所述食品可以是含有本发明呈味肽的食品。因此,本发明也包括本发明呈味肽在为食品提供浓厚味、鲜味、咸味、甜味和/或酸味,尤其是酸味和/或鲜味中的用途。
下文将以具体实施例的方式阐述本发明。应理解,这些实施例仅仅是示例性的,并非限制本发明的范围。除非另有说明,实施例中所采用的方法、试剂等都是本领域常规使用的方法和试剂。本文中,若无特别指出,百分含量指的是重量百分数。
实施例1:人工合成氨基酸序列为Ser-Pro-Asn和Ala-Leu-Asn的鲜味呈味肽
委托吉尔生化(上海)有限公司合成本发明鲜味呈味肽。
采用HPLC检测两种鲜味呈味肽合成物纯度,分别如图1A和1B所示,经鉴定合成物纯度依次为98.34%和98.29%。
采用UPLC-Q-TOF-MS/MS鉴定合成物分子量及结构,结果如图2A和2B所示,两种呈味肽合成物分子量约为317Da,氨基酸序列依次为Ser-Pro-Asn和Ala-Leu-Asn。
实施例2:采用滋味稀释分析方法(taste dilution analysis,TDA)评价Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽的呈味特性、TDA系数、阈值
滋味稀释分析(TDA)法如下:取合成冻干的Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽配置成5g/L的评价溶液,其基础溶液采用纯水,依次按照1:1的比例用纯水对评价溶液进行逐步稀释,逐步稀释后的评价溶液按照浓度递减的顺序呈给经过训练的评价人员(6名:3男,3女,年龄在25-45岁间),每个稀释水平溶液采用三角实验测定法进行评定。当某个稀释水平的溶液与空白之间的味觉差异不能被任何一位评价人员识别出来时,记录上一次样品稀释浓度,即为样品在该空白溶液中的阈值。此时的稀释倍数即稀释因子系数(TDA系数)。阈值及TDA系数的最终结果值取各评价人员评定结果的平均值。每份样品在不同时间内重复三次,均为常温下评定。每位感官人员还需要评价每次呈上样品的呈味特性。记录结果如表1所示。
表1:Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽的呈味特性、TDA系数、阈值
呈味肽 | 呈味特性(5g L<sup>-1</sup>) | TDA系数 | 阈值(mmol L<sup>-1</sup>) |
Ser-Pro-Asn | 酸味、少许鲜味 | 128 | 0.12 |
Ala-Leu-Asn | 酸味、鲜味 | 64 | 0.25 |
实施例3:Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽用于调味汁的效果
在市售调味汁中分别以5g/L的浓度添加Ser-Pro-Asn和Ala-Leu-Asn序列呈味肽,评价其呈味贡献。评价方法如下:样品的滋味评价分为甜味、酸味、苦味、咸味、鲜味,对所评样品的每个滋味进行0(not detectable)到5(strongly detectable)的5分制打分,结果以蜘蛛图3表示。
图3显示,本发明呈味肽对调味汁鲜味有显著增强,同时酸味也略有增强。
Claims (17)
1.一种三肽,其特征在于,所述三肽选自:
(1)Ser-Pro-Asn;和
(2)Ala-Leu-Asn。
2.如权利要求1所述的三肽,其特征在于,所述三肽能提供鲜味和/或酸味。
3.一种组合物,所述组合物含有三肽,其中,所三肽为:三肽Ser-Pro-Asn,或三肽Ala-Leu-Asn,或三肽Ser-Pro-Asn与三肽Ala-Leu-Asn。
4.如权利要求3所述的组合物,其特征在于,所述组合物是调味品。
5.如权利要求4所述的组合物,其特征在于,所述调味品为鲜味调味品或酸味调味品。
6.如权利要求5所述的组合物,其特征在于,所述调味品选自酱料、酱油、奶酪涂抹料、汤料和味精。
7.如权利要求4所述的组合物,其特征在于,所述调味品还含有5’-肌苷酸盐、5’-鸟苷酸盐、琥珀酸及其盐、植物水解蛋白、酵母抽提物、N1-(2,4-二甲氧基苄基)-N2-(2-(2-吡啶基)乙基)草酰胺、鲜味氨基酸、鲜味呈味肽和厚味呈味肽中的一种或任意多种的混合物。
8.如权利要求7所述的组合物,其特征在于,所述厚味呈味肽为谷胱甘肽和谷缬甘肽。
9.如权利要求3所述的组合物,其特征在于,所述组合物是食品。
10.如权利要求9所述的组合物,其特征在于,所述食品选自发酵食品、保健食品、传统面制品和烘焙食品。
11.如权利要求3所述的组合物,其特征在于,
所述组合物是选自油脂产品、饮料、汤、冰激凌、酸奶、蛋糕调和粉和奶酪的食品。
12.如权利要求3-11中任一项所述的组合物,其特征在于,所述组合物中,所述三肽的含量为0.01~20重量%。
13.如权利要求12所述的组合物,其特征在于,所述组合物中,所述三肽的含量为0.1~15重量%。
14.一种增加食物鲜味和/或酸味的方法,所述方法包括向所述食物中加入权利要求1-2中任一项所述的三肽,或权利要求3-8中任一项所述的组合物。
15.如权利要求14所述的方法,其特征在于,所述组合物中所述三肽的含量为0.01~20重量%。
16.如权利要求15所述的方法,其特征在于,所述组合物中所述三肽的含量为0.1~15重量%。
17.权利要求1-2中任一项所述的三肽在增加食物或调味品鲜味和/或酸味中的应用,或在制备用于增加食物或调味品鲜味和/或酸味的调味品或食品中的应用。
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US7148192B2 (en) * | 2004-01-29 | 2006-12-12 | Ebwe Pharma Ges. M.H. Nfg.Kg | Neuroprotective dietary supplement |
JP5892687B2 (ja) * | 2011-07-07 | 2016-03-23 | 高砂香料工業株式会社 | 風味改善ペプチド |
CN102396688B (zh) * | 2011-11-02 | 2013-05-08 | 华南理工大学 | 呈味活性肽及其制备方法与应用 |
CN102603870B (zh) * | 2012-04-10 | 2013-06-12 | 上海海洋大学 | 一种呈味肽及其制备方法和应用 |
CA2896246C (en) * | 2013-01-22 | 2021-07-06 | Mars, Incorporated | Flavor composition and edible compositions containing same |
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