CN108610395A - 一种具有鲜味和增鲜特性的多肽及其用途 - Google Patents
一种具有鲜味和增鲜特性的多肽及其用途 Download PDFInfo
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Abstract
本发明公开了一种具有鲜味和增鲜特性的四肽及其用途,该四肽的一级结构(氨基酸序列)为Gly‑Gly‑Asn‑Pro(GGNP)。本发明的鲜味/增鲜四肽可以L‑氨基酸为原料,参照现有技术的化学方法合成(比如固相合成法);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/增鲜四肽具有较好的鲜味和增鲜特性,可有效的提升味精、酱油及鸡肉汤的鲜味强度、浓郁感以及饱满感,可应用于速冻食品、调味品、饮料和肉汤等多种食品当中。
Description
技术领域
本发明属于食品添加剂领域,具体涉及一种具有鲜味和增鲜特性的四肽及其用途,该四肽添加于速冻食品、调味品(如酱油、醋)、饮料以及肉汤等多种食品中,可有效地提升其鲜味和饱满感。
背景技术
味精作为第一代鲜味剂在20世纪初就实现了工业化生产,并随后在全世界得到广泛应用,但由于其易引起口干,在高温下易形成有毒性的焦化谷氨酸钠而日益淡出调味品市场。近年来涌现出来的新型天然鲜味剂有着不可比拟的优势,它们既能满足消费者的味感需求,也符合消费者重视产品天然属性和环保属性的心理需求。呈味肽是这些新兴天然鲜味剂的重要组成部分,不仅呈现不同于味精的复杂鲜味,同时还能参与并影响食品风味的形成,提高食品风味。
呈味肽自被发现以来,因其安全性好和化学稳定性高、呈味阈值低、醇厚味长、易消化吸收等特点而成为研究热点。呈味肽因肽链的长度、氨基酸组成、排列顺序不同而呈现不同的味感。例如有学者对生猪肉和干熏火腿肉抽提物进行分离纯化,发现分子量在1500~1700Da的肽段具有明显鲜味;还有人研究发现小于500Da的酸性小肽组分是酱油中的主要鲜味成分。
中国发明专利ZL 201510269498.6公开了一种牛肝菌美拉德鲜味肽及其制备方法,该鲜味肽可用作食品添加剂,还可作为新型天然复合调味料的基础原料。
中国发明专利申请(申请号201710658647.7)公开了一种具有鲜味的五肽,该五肽添加于速冻食品、酱油、醋、饮料、肉汤等多种食品中或调味料等多种可食用材料中,将有效地提升其鲜味。
目前鲜味肽的生产主要以动植物或微生物蛋白为原料通过酶解法制备得到,但是得到的蛋白酶解产物复杂多样,且目标肽段含量较少,并在生产过程中伴随着苦味肽的生成,因此如何在鲜味肽的制备过程中得到具有良好风味的鲜味肽同时降低生产成本也是食品工业亟待解决的关键问题。
发明内容
本发明的首要目的在于提供一种具有鲜味和增鲜特性的四肽。
本发明的另一目的在于提供上述四肽的用途。
本发明的目的通过下述技术方案实现:
一种四肽,其一级结构氨基酸序列为Gly-Gly-Asn-Pro(GGNP),其中Gly为甘氨酸,Asn为天冬酰胺,Pro为脯氨酸,所有氨基酸均为L构型。
上述四肽的可食用的盐类物质也具有增鲜特性。
上述四肽可以采用现有技术的化学方法合成,基于一般性的化学合成思路,选择较佳的合成路线,可以获得较高的收率。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。
此外本发明的鲜味/增鲜四肽也可采用醇沉、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液美拉德反应产物中分离获得。
本发明的四肽或其可食用的盐类物质可以作为增鲜类食品添加剂使用,添加量为食品质量的0.01-1.00%,根据具体食品体系或食品工艺的要求确定。
本发明相对于现有技术具有如下的优点及效果:
本发明四肽具有较好的鲜味和较好的增鲜特性,在低浓度范围内(0-2mmol/L)具有强于味精的增鲜特性,即本发明四肽在低于2mmol/L的范围内能与其他鲜味剂发生协同增效作用使得体系最终鲜味强度优于同等味精添加量下的食品体系。除此之外,本发明四肽与其他呈味物质还具有协同增效作用从而能显著提升味精溶液、鸡肉汤或酱油等食品的饱满感、持久感以及醇厚感,赋予食品体系多种层次的味感。
附图说明
图1为Sephadex G-25凝胶过滤色谱分离图谱。
图2为超高效液相色谱分离图谱。
图3为四肽Gly-Gly-Asn-Pro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四肽进行感官评价。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;
(3.1)取99ml味精溶液(0.05%)加入1ml蒸馏水作为对照组,另取99ml味精溶液(0.05%)加入1ml制备好的多肽作为样品;
(3.2)取99ml酱油加入1ml蒸馏水作为对照组,另取99ml酱油加入1ml制备好的多肽作为样品;
(3.3)取49ml鸡肉汤加入1ml蒸馏水作为对照组,另取49ml鸡肉汤加入1ml制备好的多肽作为样品。
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)组成,这些成员对五种基本味觉品质十分熟悉。感官评价房间里的温度控制在23±2℃。评价样品时采用评分法,从0分到10分,0分表示被检测的样品没有味道,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Gly-Gly-Asn-Pro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Gly-Gly-Asn-Pro粗品,用反相高效液相色谱法纯化,得到多肽纯品。
实施例2
本发明多肽的分离纯化方法,包括以下步骤:
(1)花生粕曲料的制备:取花生粕,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;
(2)花生粕酶解物的制备: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;
(3)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。
(4)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。选择鲜味最强的组分F5进行冷冻干燥,得到目标多肽。目标多肽的二级质谱图如图3所示,序列是Gly-Gly-Asn-Pro。
表1各实施例感官分析
由表1可以看出,本发明所述的多肽具有较好的鲜味和增鲜特性,其鲜味阈值为2.10mmol/L,而增鲜阈值则仅为0.63mmol/L,说明当本发明所述多肽浓度高于0.63mmol/L时便能显著增强味精溶液的鲜味强度。
从表2我们可以得知,本发明所述多肽还能够显著提升食品(如味精溶液、酱油和鸡肉汤)的鲜味、饱满感,让食品整体口感的鲜味和浓郁特性得到进一步提升,说明此发明多肽是一种较好的鲜味剂替代品。
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)
1.一种四肽,其特征在于:一级结构氨基酸序列为Gly-Gly-Asn-Pro。
2.权利要求1所述的四肽或其可食用的盐类物质作为增鲜类食品添加剂的应用。
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