CN111557343A - 一种抗食物过敏和增强机体免疫功能的豆奶饮料及其制备方法 - Google Patents
一种抗食物过敏和增强机体免疫功能的豆奶饮料及其制备方法 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C2240/00—Use or particular additives or ingredients
- A23C2240/15—Use of plant extracts, including purified and isolated derivatives thereof, as ingredient in dairy products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/113—Acidophilus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/137—Delbrueckii
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明涉及一种抗食物过敏和增强机体免疫功能的豆奶饮料及其制备方法,属于食品饮料加工技术领域,由以下物质按质量百分比组成:乳酸菌发酵豆奶50~55%、马齿苋提取物2.0~3.0%、葡萄籽提取物1.5~2.0%、橘皮果胶0.2~0.5%、蔗糖1.5~2.0%、低聚糖4.0~5.0%、维生素C 0.03~0.05%、余量为水。抗食物过敏和增强免疫作用的豆奶饮料由大豆经发芽、打浆、煮浆、乳酸菌发酵和添加其它抗过敏活性成分等工序制成。本发明的豆奶饮料含有多种抗过敏成分,抗过敏效果显著;同时,其营养丰富,风味口感好,具有提高机体的免疫调节功能;且制备方法简单,饮料的稳定好,可适合工业化生产。
Description
技术领域
本发明属于食品饮料加工技术领域,特别涉及一种抗食物过敏和增强机体免疫功能的豆奶饮料及其制备方法。
背景技术
食物过敏是机体暴露于特定食物时可以重复发生的危害机体健康的特定免疫反应。食物过敏的患病率在近几十年内急剧上升,患者数量以百万计,食物过敏已成为一个全球性的健康问题。目前,推荐的食物过敏治疗方法仍以严格禁止接触过敏原为主。但医学上能明确确定的过敏原种类有限,对于未明过敏原的过敏反应,仍是防不胜防;同时,由于引起食物过敏的过敏原绝大多数为蛋白质,这对正在发育中的小儿来说,禁止食用这些蛋白质会造成营养不良,从而影响过敏患者的生活质量。因此,人们越来越迫切地希望能找到治疗乃至根治食物过敏的方法。
食物过敏的传统治疗方法是采用药物治疗,一般采用抗组胺剂、类固醇等抗过敏药品,但这些药品往往存在较大的副作用,常见不良反应有头晕、嗜睡、口干恶心等,且长期使用容易产生耐药性。因此,研究开发天然、安全和无毒副作用的抗过敏食品和药品具有重要的意义。目前,从动植物中提取天然活性物质用于预防或治疗食物过敏性疾病,具有材料来源广泛、毒副作用小、可针对多种过敏食物等优点,这是化学药物抗过敏作用所无法比拟的。
近年来,越来越多的证据表明机体内微生物的生态失调是引起食物过敏的主要原因,因此乳酸菌等益生菌对过敏症状的潜在预防和治疗效果引起了人们的广泛关注。大量研究发现乳酸菌不仅能够通过发酵降低食物过敏原的致敏性,而且还能通过调节肠道菌群的平衡在宿主免疫***的发育和调节中发挥作用,从而改变过敏性疾病的发病风险。
迄今为止,开展的基于天然产物的抗过敏研究大多针对某一抗过敏成分,但是其抗过敏作用十分有限,不足以缓解过敏症状的程度。因此,利用包括益生菌等在内的多种抗过敏组分的协同增效作用,使得组合物的抗过敏作用各司其责,呈现出多层次、多靶点的特点,能够在抑制免疫球蛋白IgE产生、对抗过敏介质、中和变应原等多个环节中起作用,这将是未来抗过敏食品的发展方向。
发明内容
针对现有技术中存在的问题,本发明的目的在于提供一种抗食物过敏和增强机体免疫功能的豆奶饮料及其制备方法。
为解决上述问题,本发明采用如下的技术方案。
一种抗食物过敏和增强免疫作用的豆奶饮料,由以下物质按质量百分比组成:乳酸菌发酵豆奶50~55%、马齿苋提取物2.0~3.0%、葡萄籽提取物1.5~2.0%、橘皮果胶0.2~0.5%、蔗糖1.5~2.0%、低聚糖4.0~5.0%、维生素C 0.03~0.05%、余量为水。
所述豆奶饮料的制备方法,包括如下步骤:
(1)将大豆经水清洁,用水浸泡8~10h,大豆转移至种子培养箱中,湿度为85%,培养温度为28~32℃,发芽时间为3~5天;
(2)将得到的大豆芽冲洗干净,与水按重量比1:8的配比进行磨浆,煮沸10~15分钟后,用100目筛过滤除去豆渣,得发芽豆浆;
(3)制备乳酸菌发酵豆奶:向步骤(2)发芽豆浆中添加蔗糖,经均质、灭菌和冷却后,在无菌操作下接种乳酸菌发酵菌剂,接菌量3~5%,在37~42℃下恒温发酵24~36h,直至豆奶pH值为3.7~4.0时,终止发酵,将发酵产物进行冷却处理,得到乳酸菌发酵豆奶;
所述的乳酸菌发酵菌剂包括嗜酸乳杆菌CICC 6081、植物乳杆菌植物亚种CICC20988和德式乳杆菌保加利亚亚种CICC 6103的单菌种或它们的混合菌种;
(4)按比例将低聚糖、果胶、葡萄籽提取物和维生素C加入到马齿苋提取物中,再添加一部分水进行搅拌混合后,进行微波杀菌处理;
所述马齿苋提取物的制备:取马齿苋全草,经干燥、粉碎后过100目筛,采用5~8倍60%乙醇,在60℃水浴条件提取,经过滤、干燥得到;
(5)按比例将步骤(3)中乳酸菌发酵豆奶加入步骤(4)的杀菌产物中,再加入剩余的无菌水,充分混合,无菌罐装,即得抗过敏营养豆奶饮料。
本发明将不同种类的乳酸菌与Balb/c小鼠脾细胞在体外进行共培养,并以合适的卵白蛋白剂量刺激小鼠脾细胞致敏,以脾细胞分泌Th2型细胞因子IL-4及Th1型细胞因子IFN-γ的水平选择三种抗过敏作用显著的乳酸菌作为发酵菌剂,通过接种发芽豆浆,再添加其他天然抗过敏因子制备抗食物过敏和增强机体免疫功能的豆奶饮料;并利用构建的KU812细胞脱颗粒模型,体外评估本发明豆奶饮料的抗过敏作用。
本发明与现有技术相比具有显著的优点和有益效果,具体表现在:
(1)具有显著的抗过敏功能。①制备饮料所用的乳酸菌活菌是经过免疫调节实验筛选获得,饮料中含有大量的乳酸菌活菌、马齿苋提取物、葡萄籽提取物和橘皮果胶都具有抗过敏功效;同时,经发酵后的乳酸菌豆奶饮料中游离的异黄酮增加,这些成分协同作用产生显著的抗过敏效果。②大豆经发芽、打浆和乳酸菌发酵后,饮料中大豆蛋白降解为具有免疫功能的活性肽;饮料中所含低聚糖也可以通过增殖肠道中的乳酸菌,其在平衡肠道菌群、改善胃肠道和消化***疾病方面效果明显,长期服用能强身健体,增强机体的免疫调节功能。因此,本发明饮料特别是适用于食物过敏患者食用,为抗过敏预防提供了全新的选择和策略。
(2)具有高的营养价值。大豆经发芽、乳酸菌发酵后总氨基酸、维生素C及铁、钙、锌和磷等矿物质含量显著增加,并降低了抗营养因子等不利于健康的因素;同时,乳酸菌发酵赋予产品风味口感好,无豆腥味。
(3)制备方法简单,易于实际操作,果胶还可作为良好的稳定剂解决了饮料的沉淀和分层问题,本发明饮料采用纯天然成分,安全性好,生产成本低,可适合工业化生产,市场前景广阔。
具体实施方式
实施例1
一种抗食物过敏和增强免疫作用的豆奶饮料,由以下物质按质量百分比组成:乳酸菌发酵豆奶50%、马齿苋提取物2.0%、葡萄籽提取物1.5%、橘皮果胶0.2%、蔗糖1.5%、低聚糖4.0%、维生素C 0.03%、余量为水。
所述豆奶饮料的制备方法,包括如下步骤:
(1)将大豆经水清洁,用水浸泡8h,大豆转移至种子培养箱中,湿度为85%,培养温度为28℃,发芽时间为3天;
(2)将得到的大豆芽冲洗干净,与水按重量比1:8的配比进行磨浆,煮沸10分钟后,用100目筛过滤除去豆渣,得发芽豆浆;
(3)制备乳酸菌发酵豆奶:向步骤(2)发芽豆浆中添加蔗糖,经均质、灭菌和冷却后,在无菌操作下接种乳酸菌发酵菌剂,接菌量3%,在37℃下恒温发酵24h,直至豆奶pH值为3.7时,终止发酵,将发酵产物进行冷却处理,得到乳酸菌发酵豆奶;
所述的乳酸菌发酵菌剂包括嗜酸乳杆菌CICC 6081、植物乳杆菌植物亚种CICC20988和德式乳杆菌保加利亚亚种CICC 6103的单菌种或它们的混合菌种;
(4)按比例将低聚糖、果胶、葡萄籽提取物和维生素C加入到马齿苋提取物中,再添加一部分水进行搅拌混合后,进行微波杀菌处理;
所述马齿苋提取物的制备:取马齿苋全草,经干燥、粉碎后过100目筛,采用5倍60%乙醇,在60℃水浴条件提取,经过滤、干燥得到;
(5)按比例将步骤(3)中乳酸菌发酵豆奶加入步骤(4)的杀菌产物中,再加入剩余的无菌水,充分混合,无菌罐装,即得抗过敏营养豆奶饮料。
实施例2
一种抗食物过敏和增强免疫作用的豆奶饮料,由以下物质按质量百分比组成:乳酸菌发酵豆奶52%、马齿苋提取物2.5%、葡萄籽提取物1.7%、橘皮果胶0.3%、蔗糖1.8%、低聚糖4.5%、维生素C 0.04%、余量为水。
所述豆奶饮料的制备方法,包括如下步骤:
(1)将大豆经水清洁,用水浸泡8h,大豆转移至种子培养箱中,湿度为85%,培养温度为30℃,发芽时间为4天;
(2)将得到的大豆芽冲洗干净,与水按重量比1:8的配比进行磨浆,煮沸12分钟后,用100目筛过滤除去豆渣,得发芽豆浆;
(3)制备乳酸菌发酵豆奶:向步骤(2)发芽豆浆中添加蔗糖,经均质、灭菌和冷却后,在无菌操作下接种乳酸菌发酵菌剂,接菌量4%,在40℃下恒温发酵30h,直至豆奶pH值为3.8时,终止发酵,将发酵产物进行冷却处理,得到乳酸菌发酵豆奶;
所述的乳酸菌发酵菌剂包括嗜酸乳杆菌CICC 6081、植物乳杆菌植物亚种CICC20988和德式乳杆菌保加利亚亚种CICC 6103的单菌种或它们的混合菌种;
(4)按比例将低聚糖、果胶、葡萄籽提取物和维生素C加入到马齿苋提取物中,再添加一部分水进行搅拌混合后,进行微波杀菌处理;
所述马齿苋提取物的制备:取马齿苋全草,经干燥、粉碎后过100目筛,采用7倍60%乙醇,在60℃水浴条件提取,经过滤、干燥得到;
(5)按比例将步骤(3)中乳酸菌发酵豆奶加入步骤(4)的杀菌产物中,再加入剩余的无菌水,充分混合,无菌罐装,即得抗过敏营养豆奶饮料。
实施例3
一种抗食物过敏和增强免疫作用的豆奶饮料,由以下物质按质量百分比组成:乳酸菌发酵豆奶55%、马齿苋提取物3.0%、葡萄籽提取物2.0%、橘皮果胶0.5%、蔗糖2.0%、低聚糖5.0%、维生素C 0.05%、余量为水。
所述豆奶饮料的制备方法,包括如下步骤:
(1)将大豆经水清洁,用水浸泡10h,大豆转移至种子培养箱中,湿度为85%,培养温度为30℃,发芽时间为5天;
(2)将得到的大豆芽冲洗干净,与水按重量比1:8的配比进行磨浆,煮沸15分钟后,用100目筛过滤除去豆渣,得发芽豆浆;
(3)制备乳酸菌发酵豆奶:向步骤(2)发芽豆浆中添加蔗糖,经均质、灭菌和冷却后,在无菌操作下接种乳酸菌发酵菌剂,接菌量5%,在42℃下恒温发酵36h,直至豆奶pH值为4.0时,终止发酵,将发酵产物进行冷却处理,得到乳酸菌发酵豆奶;
所述的乳酸菌发酵菌剂包括嗜酸乳杆菌CICC 6081、植物乳杆菌植物亚种CICC20988和德式乳杆菌保加利亚亚种CICC 6103的单菌种或它们的混合菌种;
(4)按比例将低聚糖、果胶、葡萄籽提取物和维生素C加入到马齿苋提取物中,再添加一部分水进行搅拌混合后,进行微波杀菌处理;
所述马齿苋提取物的制备:取马齿苋全草,经干燥、粉碎后过100目筛,采用8倍60%乙醇,在60℃水浴条件提取,经过滤、干燥得到;
(5)按比例将步骤(3)中乳酸菌发酵豆奶加入步骤(4)的杀菌产物中,再加入剩余的无菌水,充分混合,无菌罐装,即得抗过敏营养豆奶饮料。
抗过敏实例
1.KU812细胞的培养
KU812细胞为悬浮细胞,于配置好的1640完全培养基中培养,放置于37℃、5%CO2的培养箱孵育。每日观察细胞长势,采用半换液方式,每隔两天加入新鲜完全培养基,当细胞密度占全部培养基80%左右时进行传代。
2.诱导KU812细胞脱颗粒模型
(1)取状态良好的KU812细胞,常温离心,弃上清,加入含0.5μg/mL Human MyelomaIgE(人骨髓瘤IgE)的完全培养基,于37℃、5%CO2的培养箱孵育一周,以促进KU812细胞表面FcεRI受体的表达。
(2)将处于对数期、形态完整的细胞分别接种到24孔细胞培养板中(1mL/孔),培养过夜使细胞适应新的环境。要求每孔KU812细胞数目达到2×106/mL。
(3)次日,每孔加入20μL鸡蛋过敏患者混合血清与细胞进行孵育,37℃、5%CO2继续培养24h。
(4)实验组每孔加入加入50μL乳酸菌发酵豆奶饮料或对照样品,培养1h;空白对照组加入50μL的无菌PBS溶液。
(5)向各组中加入20μL卵白蛋白(20μg/孔)激发细胞,培养4h,常温下离心,收集上清液用于检测或分装置于-20℃冻存备用。
3.β-氨基己糖苷酶(β-HEX)释放率
待细胞被卵白蛋白激发4h后,取出每组细胞并分成实验组和对照组两组进行检测。取细胞液离心后的上清液作为实验组;同时,在细胞液中加入TritonX-100,裂解细胞30min,离心后取上清液作为对照组。取每组的上清50μL,并以50μL PBS为空白对照,在每孔中加50μL的底物(4-硝基苯-N-乙酰-β-D-氨基葡萄糖苷),放置37℃培养箱反应1h后,加入200μL终止液(0.1mol/L Na2CO3/NaHCO2)终止反应。于405nm处测OD值,根据各组所测OD值,按照下列公式计算β-HEX释放率。
4.组胺释放量
细胞待卵白蛋白激发4h后,离心,收集细胞上清液,利用商业化人组胺ELISA试剂盒检测细胞激发后释放的组胺含量。
5.细胞因子分泌水平
细胞待卵白蛋白激发4h后,离心,收集上清液,利用商业化ELISA试剂盒检测细胞激发后培养上清液中IL-4及IFN-γ的含量。
表1各组抗过敏效果
为了评估本发明豆奶饮料的抗过敏效果,将发明中使用的3个抗过敏组分乳酸菌溶液、马齿笕提取物、发酵豆奶与发明实施例样品在相同条件进行抗过敏效果评估,从表1可看出,单独使用三种抗过敏组分,与空白对照组相比,乳酸菌溶液、发酵豆奶、马齿笕提取物与致敏KU812细胞孵育后能够显著提高细胞分泌IFN-γ的水平,降低细胞分泌IL-4的水平,即增强Th1免疫、抑制Th2免疫,同时β-HEX和组胺释放率显著下降,具有抗食物过敏的作用。但本发明饮料与其组分相比,进一步增强了IFN-γ的分泌,抑制IL-4的产生,β-HEX和组胺释放率继续下降,说明本发明的豆奶饮料在多种抗过敏成分协同作用下,抗过敏作用更加显著。
以上所述仅表达了本发明的优选实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形、改进及替代,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (4)
1.一种抗食物过敏和增强免疫作用的豆奶饮料,其特征是由以下物质按质量百分比组成:乳酸菌发酵豆奶50~55%、马齿苋提取物2.0~3.0%、葡萄籽提取物1.5~2.0%、橘皮果胶0.2~0.5%、蔗糖1.5~2.0%、低聚糖4.0~5.0%、维生素C 0.03~0.05%、余量为水。
2.根据权利要求1所述豆奶饮料的制备方法,其特征是包括如下步骤:
(1)将大豆经水清洁,用水浸泡8~10h,大豆转移至种子培养箱中,湿度为85%,培养温度为28~32℃,发芽时间为3~5天;
(2)将得到的大豆芽冲洗干净,与水按重量比1:8的配比进行磨浆,煮沸10~15分钟后,用100目筛过滤除去豆渣,得发芽豆浆;
(3)制备乳酸菌发酵豆奶:向步骤(2)发芽豆浆中添加蔗糖,经均质、灭菌和冷却后,在无菌操作下接种乳酸菌发酵菌剂,接菌量为3~5%,在37~42℃下恒温发酵24~36h,直至豆奶pH值为3.7~4.0时,终止发酵,将发酵产物进行冷却处理,得到乳酸菌发酵豆奶;
(4)按比例将低聚糖、果胶、葡萄籽提取物和维生素C加入到马齿苋提取物中,再添加一部分水进行搅拌混合后,进行微波杀菌处理;
(5)按比例将步骤(3)中乳酸菌发酵豆奶加入步骤(4)的杀菌产物中,再加入剩余的无菌水,充分混合,无菌罐装,即得抗过敏营养豆奶饮料。
3.根据权利要求2所述豆奶饮料的制备方法,其特征是,所述的乳酸菌发酵菌剂包括嗜酸乳杆菌CICC 6081、植物乳杆菌植物亚种CICC 20988或德式乳杆菌保加利亚亚种CICC6103的单菌种或它们的混合菌种。
4.根据权利要求2所述豆奶饮料的制备方法,其特征是,所述马齿苋提取物的制备:取马齿苋全草,经干燥、粉碎后过100目筛,采用5~8倍60%乙醇,在60℃水浴条件提取,经过滤、干燥得到。
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