CN108634280A - 一种鲜味六肽及其用途 - Google Patents
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Abstract
本发明公开了一种鲜味六肽及其应用,该多肽的一级结构氨基酸序列为TESSSE。本发明的鲜味六肽可以L‑氨基酸为原料,参照现有技术的化学方法合成;也可采用醇沉、葡聚糖凝胶色谱、反相高效液相色谱等分离手段从花生蛋白发酵酶解产物中分离获得。本发明的鲜味六肽鲜味阈值小于味精鲜味阈值,具有较好的鲜味及增鲜特性,可显著提升味精、酱油及鸡肉汤的鲜味强度,并不同程度提升味精、酱油和鸡肉汤的饱满感、持久感以及醇厚感,让其整体口感的鲜味、圆润以及浓郁特性得到进一步提升。
Description
技术领域
本发明属于食品添加剂领域,具体涉及一种鲜味六肽及其应用。
背景技术
鲜味是继酸、甜、苦、咸之后的第五个基本味觉。最早被报道的鲜味物质是1908年Ikeda在海白菜中首次发现的谷氨酸钠(MSG),科学家将其滋味特性命名为“umami”,随后一些呈味核苷酸(IMP、GMP)也被发现具有鲜味。1978年Yamasaki等人采用凝胶过滤和离子交换色谱从牛肉蛋白酶解液中分离纯化获得美味肽,并鉴定其一级结构为Lys-Gly-Asp-Glu-Ser-Leu-Leu-Ala,随后科学家将类似于美味肽的具有鲜味的肽类物质统一命名为鲜味肽,目前为止,国内外学者对鲜味肽进行了大量研究。
鲜味肽可通过动植物蛋白酶解和发酵获得,结合膜分离、凝胶色谱、高效液相色谱等技术对其进行分离纯化可得到更高纯度的鲜味肽。鲜味肽应用于食品中符合“天然、营养、安全”的食品发展潮流,越来越受到人们的青睐。鲜味肽在食品风味中发挥的作用并不局限于其对鲜味的贡献,在增强食品其他风味或提供风味前提物质进行美拉德反应产生目标风味方面也具有不可比拟的优势。将鲜味肽应用于方便面、调味品、肉制品、酱腌菜等食品调味中,不仅可降低盐的咸感、味精的干涉感,还能增强产品的鲜味、肉感、浓厚感,延长后味,以及增强食品风味协调性。与其他鲜味剂相比,鲜味肽具有呈味阈值低、风味浓郁等优势,一方面能够丰富鲜味剂的品种,开辟调味品新市场;另一方面,鲜味剂也具肽类物质的生理活性功能,可以满足不同人群的需求。
发明内容
本发明的首要目的在于提供一种鲜味六肽,该肽具有明显的鲜味,且对味精及其他调味品具有较强的增鲜作用。
本发明的另一目的在于提供上述鲜味六肽的用途。
本发明的目的通过下述技术方案实现:
一种多肽(六肽),一级氨基酸序列结构式为TESSSE;其中T代表苏氨酸,E代表谷氨酸,S代表丝氨酸,所有氨基酸均为L构型。
本发明的鲜味六肽可以L-氨基酸为原料,参照现有技术的化学方法合成;在树脂上逐个偶联氨基酸序列,以形成本发明鲜味六肽。比如在固相合成仪上合成,将1g二氯树脂溶胀、洗涤,脱除Fmoc保护基后加入氨基酸进行缩合反应,重复脱保护——缩合的过程直至所有氨基酸连接完毕。
本发明的鲜味六肽也可采用醇沉、葡聚糖凝胶色谱、反相高效液相色谱等分离手段从花生蛋白发酵酶解产物中分离获得。
本发明的多肽可以作为增鲜类食品添加剂使用,用于提升食品的鲜味和饱满度。
本发明相对于现有技术具有如下的优点及效果:
本发明的鲜味六肽鲜味阈值低于味精鲜味阈值,具有较好的鲜味及增鲜特性,不仅可显著提升味精、酱油及鸡肉汤的鲜味强度,而且可不同程度提升味精、酱油和鸡肉汤的饱满感、持久感以及醇厚感,让其整体口感的鲜味、圆润以及浓郁特性得到进一步提升。
附图说明
图1为花生蛋白酶解物的Sephadex G-25凝胶过滤色谱分离图谱。
图2为花生鲜味肽的超高效液相色谱分离图谱。
图3为花生鲜味六肽TESSSE的二级质谱图。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述,但本发明的实施方式不限于此。
以下实施例中,各感官品评的测定方法如下:
(1)鲜味阈值测定方法
采用滋味稀释分析法对鲜味肽进行感官评价。将鲜味肽准确溶于纯净水中,配成1%的浓度,然后进行1:1(体积比)的逐步稀释,样品的逐步稀释的溶液按照浓度增加的顺序呈给经过训练的评员,每个稀释水平溶液采用3点测定进行评定。当某个稀释水平的溶液与2个空白(纯净水)之间的滋味差异刚好能被识别出来,那么称这时的样品浓度为可识别的阈值。每个评定组的阈值采用各个评员评定结果的平均值。
(2)增鲜阈值测定方法
采用比较滋味稀释分析法对鲜味六肽进行感官评价。将滋味稀释分析法中的空白(纯净水)换为0.03mg/L的味精溶液,其他步骤如鲜味阈值测定方法。
(3)鲜味肽在食品和调味品中的应用
取各实施例中制得的多肽制备成10mg/ml的溶液;
①取1mL蒸馏水加入99mL味精溶液(0.05%)中作为对照组,另取1mL制备好的多肽加入99mL味精溶液(0.05%)中作为样品;
②取1mL蒸馏水加入99mL酱油中作为对照组,另取1mL制备好的多肽加入99mL酱油中作为样品;
③取1mL蒸馏水加入49mL鸡肉汤中作为对照组,另取1mL制备好的多肽加入49mL鸡肉汤中作为样品。
感官评价小组由10个小组成员(五男五女,年龄在25岁至40岁之间)组成,这些成员对五种基本味觉十分熟悉。感官评价房间内温度控制在23±2℃。评价样品时采用评分法,从0分到10分,0分表示被评价的样品没有味道,而10分则表示样品具有显著的味道。选择对照组作为标准品鲜味评分为5分,不同样本在这些标准品基础上进行打分评估,最后绘制图表。
实施例1
固相合成法合成TESSSE多肽,包括以下步骤:
将1g二氯树脂溶胀、洗涤,脱除Fmoc保护基后加入氨基酸进行缩合反应,重复脱保护——缩合的过程直至所有氨基酸连接完毕。切割树脂,得到多肽TESSSE粗品,用反相高效液相色谱法纯化,得到鲜味六肽1。
实施例2
多肽TESSSE的分离纯化方法,包括以下步骤:
(1)花生蛋白发酵酶解物的制备:按花生粕重量加入0.8倍去离子水并混合均匀,将均匀混合后的湿润花生粕灭菌后冷却至常温,然后将菌种与面粉混合均匀,并接种至冷却好的灭菌花生粕上,搅拌均匀;在湿度为95%、温度为32℃条件下发酵48h,得花生粕发酵曲料;按重量取花生粕发酵曲料1份和去离子水8份混合均匀,在55℃的条件下酶解18小时,95℃下灭酶15min,离心,取上清液,得到花生粕酶解液;
(2)鲜味肽的分离纯化:将花生粕酶解液用不同浓度乙醇溶液进行分离分级处理(即取浓缩后的花生粕酶解液(固形物浓度40%)80mL加入20mL食用级无水乙醇后在25℃下搅拌30min,离心(8000r/min,4℃20min)得沉淀物1和上清液1,去除多余乙醇后将沉淀物1用去离子水复溶,称为20%-组分;继续往上清液1中加入食用级无水乙醇使得体系中乙醇最终浓度为40%,在同等条件下搅拌、离心分离得沉淀物2和上清液2,沉淀物2在去除多余乙醇后复溶得40%-组分;以此类推,分别得60%-组分、80%-组分,最后剩余的上清液5除去多余乙醇溶液后其水溶液形式称为上清液。),取其中的60%-组分和80%-组分,冷冻干燥成粉末;再用凝胶过滤色谱柱G-25进行分离纯化,用去离子水以0.2-2ml/min的流速进行洗脱,检测波长为214nm,洗脱曲线如图1所示,选择鲜味强度最强的组分F4进行收集,然后冷冻干燥成粉末。
(3)将步骤(3)得到的F4组分溶解后用超高效液相色谱法进一步进行纯化,色谱柱为HSS T3柱(1.8μm,2.1×100mm;Waters,USA),流动相为A相(超纯水)和B相(乙腈),洗脱程序为:0-2min内采用100%的A相,2-10min内A与B体积比为100:0-75:25,8-10min内A与B体积比为75:25-30:70,流速为0.2ml/min,检测波长为214nm,洗脱曲线如图2所示。选择鲜味强度第二强的组分F8,冷冻干燥,得到鲜味六肽2,其二级质谱图如图3所示,序列为TESSSE。
表1鲜味六肽的鲜味及增鲜阈值
表2鲜味六肽在食品中的应用效果评价
由表1可发现,本发明的鲜味六肽(实施例1和实施例2)的鲜味阈值均小于0.4mmol/L,即该鲜味六肽在浓度约0.03%下可表现出明显的鲜味感觉,较味精鲜味阈值(0.04%)更小;此外,该鲜味六肽在0.36mmol/L浓度下(即0.026%)可明显提升味精的鲜味强度。由此可见,本发明的鲜味六肽具有较好的鲜味及增鲜特性。
由表2可知,本发明的鲜味六肽(实施例1和实施例2)可显著提升味精、酱油及鸡肉汤的鲜味强度,并不同程度提升味精、酱油和鸡肉汤的饱满感、持久感以及醇厚感,让其整体口感的鲜味、圆润以及浓郁特性得到进一步提升。说明本发明的鲜味六肽在食品和调味品中具有较好的应用前景。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (2)
1.一种多肽,其特征在于一级结构氨基酸序列为TESSSE。
2.权利要求1所述的多肽作为增鲜类食品添加剂的应用。
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