CN101534658B - 基于谷物混悬液的含有经包囊益生菌的预发酵共生基质、其制备方法和相关应用 - Google Patents
基于谷物混悬液的含有经包囊益生菌的预发酵共生基质、其制备方法和相关应用 Download PDFInfo
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
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Abstract
本发明涉及基于谷物混悬液(优选燕麦)的含有经包囊益生菌和益生元的预发酵共生基质、其制备方法和相关应用。本发明的一个目的是开发利用包囊的益生菌以及游离的和/或包囊的益生元进行预发酵的谷物共生基质(优选来自燕麦)，目的是补充现有的功能性食品市场，以及解决由于益生菌活力丧失到低于促进生物活性所需之最低限度的值所导致的这些食品的货架期缩短的固有问题。此外，本发明的一个目的是不同程度地改善发酵过程条件，即缩短发酵时间以降低该过程中的能耗，降低污染的风险以及促进维持长期的微生物稳定性。所述预发酵共生基质被设计为特别地用于对乳制品不耐受和/或过敏的那些情形，然而其还适用于制药工业、化妆品工业以及优选地食品工业(包括宠物食品)。
Description
技术领域
本发明涉及含有益生元(prebiotics)和/或益生菌(probiotics)的预发酵共生基质的制备，其适用于制药工业、化妆品工业以及优选地食品(包括宠物食品)工业。所述预发酵共生基质不含或不是乳成分，适用于所有群体，尤其适用于对乳制品不耐受和/或过敏的那些个体。
发明概述
本发明的一个目的是(优选从燕麦)开发谷物共生基质，其与包囊的益生菌以及游离的和/或包囊的益生元一起进行预发酵，目的是补充现有的功能性食品市场，以及解决这些食品的货架期缩短的固有问题。此外，本发明的一个目的是不同程度地改善发酵过程条件，即缩短发酵时间以节省制备过程中的能耗，降低污染的风险以及维持长期的微生物稳定性。
背景技术
在过去几十年中，有关饮食对人类健康之影响的详细知识大大增加，全世界的人们开始意识到需要所谓的“健康饮食”，这由预期寿命延长来佐证并代表了一个重要的公共健康问题。随着益生菌产品在消费者中日益流行，食品公司需要面临制造这种产品的要求以适当地满足稳定的市场需求。一般意义上来讲，所有食品均是功能性的，只要它们提供生长和维持所必需的能量和营养。如果以有效的科学方法明确地证明及科学地证实食品成分能有益地影响健康，超出与其相关的典型营养作用，则认为该食品成分是功能性的。
该市场的特征在于其是不断变化及创新性的，其中世界水平的年市场份额为10-15％，年增长速度为20-30％。
益生菌可被定义为由于能促进宿主肠内的细菌生态系统平衡而有益地影响所述宿主的活的微生物。潜在地被认为是益生菌的乳杆菌属(Lactobacillus)、双歧杆菌属(Bifidobacterium)和肠球菌属(Enterococcus)向宿主提供免受感染的保护，认为它们阻止特定肠致病原的攻击、建立、应答和/或毒力(抗微生物活性)。这些益生菌还在控制腹泻以及降低某些形式癌症之发生的风险中具有有益的作用(抗癌活性)。还描述了降低血液胆固醇水平的作用(降胆固醇活性)。已经被科学证实的另一个可能的作用是通过产生促进乳糖消化的乳糖酶(β-半乳糖苷酶)来消化乳糖的作用，这为个体不耐受乳糖提供了解决方案。当所述益生菌产品含有最少106CFU/ml时可确保其发挥有益作用，这符合所建议的每日108-109个活细胞的最小治疗剂量的假定，其可通过摄入大约100克的每毫升或每克含有106-107个活细胞的产品来实现。由于益生菌对某些技术和功能因素(例如高水平的氧、酸性环境、冷冻以及通过胃肠道)敏感，所以其对健康的有益作用存在天然局限性。
包囊(encapsulation)方法已开始用作通过保护免受上述不利条件而提高益生菌存活率的方法。微包囊(microencapsulation)是将固体、液体或气体包装非常小的囊状物内、能在特定条件下以受控速度释放其内容物的技术。
可利用几种微包囊技术，如乳化和喷雾干燥。乳化包囊包括将含有微生物细胞和聚合物的小体积溶液(不连续相)加入到较大体积的植物油(连续相)中。然后将该混合物均质形成“油包水”乳液。一旦得到上述乳液，必须通过盐溶液使所述水溶性聚合物不溶，目的是在所述油相中形成小的凝胶颗粒。可通过改变搅拌类型及其速度以及盐溶液添加机制来控制所述囊状物的大小。乳化包囊过程易于按比例扩大并且微生物的存活率高(80-95％)。所得囊状物具有25μm-50μm的各种尺寸范围。
另一方面，所述喷雾干燥法包括利用喷雾器干燥包囊剂与活的微生物细胞的含水混合物。当溶液被汽化后与热气流(进入温度)相接触并且随后借助于真空而收集在合适容器(recipient)中从而进行干燥。该技术具有的最大优点是：步骤成本低、易于操作、可能使用对热敏感的功能性成分、所得囊状物的质量/稳定性高以及易于大量生产。所得囊状物的大小可在5-75μm之间变化。
益生元被定义为正向影响宿主、选择性刺激结肠中一种或有限几种细菌之生长和/或活性的不可消化的食品成分。
术语“共生”是指在同一食品中的具有生理活性的益生元剂和益生菌剂的协同相互作用。
目前，采用包囊形式的益生元及其与益生菌的组合吸引了食品工业的投资，其目的在于维持其长期稳定性并使相关产品的营养品质最优化。
文献US2001/0016220列举了含有可被包囊之生物活性成分的食品组分及其生产方法和应用。该文献中所述组分包括植物纤维，其包括植物纤维(包括来自燕麦的植物纤维)、可溶的和不可溶的多糖、果胶、lenhin和树胶(gum)。上述植物纤维中的生物活性组分可以是益生菌微生物、益生元、酶、营养物、次生代谢物、天然或合成的具有抗氧化活性的物质等。包囊物质可以由多糖(植物或微生物来源的)、乳化剂、肽、蛋白质和益生元物质组成。
就这些组分的获得方法而言，所讨论的文献预见到将生物活性组分引入含有形成囊状物之物质的环境中。即使该文献描述了基于谷物的、含有包囊在由谷物植物纤维形成之基质中的生物活性组分(例如益生菌和/或益生元)的产品，但是其与本发明有很大差别，因为本发明描述了预发酵谷物的含水混悬液，随后通过乳化、流化床或通过干燥来包囊微生物以及随后添加益生元组分。换句话说，在先前的文献中，所有组分(益生元或益生菌)均被包囊并被引入到相同相的基质中，而本发明描述了处于多个相中的这些组分的获得方法，其中通过包囊微生物来保护微生物的活性。
文献WO2005.002367报道了由不含益生菌微生物的燕麦制得的产品和治疗组合物，其包含蛋白质、水解蛋白和乳化脂质。除此之外，所述产品和组合物还可包含β-葡聚糖以及植物固醇。相应的生产方法通过酶促处理燕麦级分以除去碳水化合物(优选通过水解)进行。
文献WO02.065855述及由谷物分散物制得的非乳制品，其包含β-葡聚糖、蛋白质、天然糖和蛋白质。得到这样产品的方法将酶(特别是水解酶以及异构酶)施用于谷物混悬液。
文献WO02/37984报道了对基于不含大豆和奶的燕麦混悬液的微生物培养物进行发酵而得的产品以及相应的制备。该文献预见到在燕麦混悬液的发酵物中使用乳杆菌和链球菌菌株以及在之后孵育用于发酵的燕麦含水混悬液中包含几种组分，例如磷酸氢钙和/或磷酸钙、β-葡聚糖、麦芽糖、麦芽糊精、蛋白质等。
文献WO00/65930报道了由谷物(特别是燕麦)制得的进一步用作食品工业中原材料的产品。这些产品的制备方法包括由谷物的糠、薄片或粉制备混悬液。然后以预定的温度和压力对该混悬液进行均质以得到乳液。然后，可利用微生物(例如乳杆菌和双歧杆菌等)对所述乳液进行发酵、酸化和最后进行巴氏消毒(或者以粉末形式呈现)。
文献CA2383021描述了从谷物中产生的、从谷物粉或提取物中得到的、通过细菌接种用于发酵的分离自β-葡聚糖的共生组合物。乳杆菌、链球菌和/或双歧杆菌被接种在谷物的含水混悬液中，用α-淀粉酶处理，并添加稳定剂。
文献WO2004/037191描述了包含益生菌组分(例如乳杆菌和双歧杆菌)和益生元的源自大豆或乳制品的液体或冷冻形式的共生产品，其中所述益生元可由聚合物(特别是菊粉或寡果糖)组成。其制备过程使用处于液相中的益生元和益生菌组分的混合物；对该混合物进行发酵直至pH达到4.5；最后混合得到最终产品。在该最后阶段中，还可能包括一定百分比的二氧化碳。
上述6篇文献中所述的内容与本发明的内容有根本性区别，因为它们描述了仅在一相中进行燕麦混悬液添加游离的微生物或酶的发酵，并且可最终包含其它添加组分，而待分析的本发明描述了用固定化微生物预发酵谷物混悬液，随后向其加入包囊的益生菌和游离的或包囊的益生元。
当微生物以游离形式使用时，与本发明目的中利用包囊益生菌预发酵的共生基质(增加40-60％)相比，最终产品的货架期缩短，并且在贮存期间以及在通过胃肠道时微生物的稳定性/活力降低。
本发明还解决了与专利WO02.37984、WO00.65930、CA2383021、WO2004.037191中所述的货架期缩短以及维持微生物长期稳定性相关的问题(据报告与含有游离微生物的现有产品相比增加40-60％)。
文献EP 0 862 863 A2的发明目的是开发表面含有微生物和/或包封有微生物并具有可溶性纤维来源的干燥挤出谷物，列举的应用实例是早餐谷物和动物饲料。该发明的目的预见到开发用包囊益生菌及游离和/或包囊的益生元预发酵的谷物共生基质(优选在燕麦制品中)，其中当一起使用时会对最终产品中存在的微生物起到稳定作用，并且有利于通过胃肠道。此外，本发明的另一个目的是与β-葡聚糖相关的降低胆固醇的健康需求，所述β-葡聚糖作为不可消化的益生元可溶纤维的来源。所述谷物混悬液(优选燕麦制品)采用新鲜的、冻干的以及冷冻的形式，以适应食物链中介入部分的需要，因此在食品工业中有多种应用。
文献WO2004.070026公开了涉及酵母固定于κ-角叉藻聚糖或藻酸盐凝胶球中(例如在啤酒生产中)的连续方法，其通过使用静态搅拌器(static stirrer)形成乳液来实现，例如采用连续的非水相(植物油)和分散的水相(接种有酵母的κ-角叉藻聚糖)。该主题与本发明所公开的主题不同，因为尽管涉及植物油和接种有微生物之聚合物形成的乳液，但是该发明所述固定方法包含酵母，而本发明中的微生物均为益生菌微生物。
从含有加入包囊益生菌的预发酵谷物混悬液以及随后将益生元化合物引入到谷物基质中得到共生产品的分相法的构思得到优质产品，这不仅从营养角度考虑，而且还考虑了微生物长期稳定性的维持。
发明的一般描述
本发明报道了用包囊的益生菌和益生元化合物预发酵的谷物共生基质(优选燕麦制品)、其制备方法及其在多种应用(尤其在食品工业中，以及在制药工业或类似工业中)中的用途。
所得产品具有与通过传统发酵方法生产的产品相同的感官特征。
当含有包囊的微生物时，这些产品还具有如下优点，在长的货架期或在摄取后通过胃肠道期间活力/稳定性增加。
作为另一个优点，所述基质的有效期比市场上现有的产品的有效期长出40％-60％。
通过使用该技术，得到了包含残量的游离微生物并具有与传统发酵产品相同的感官特征的预发酵产品，因此是更有价值的产品。
与游离的细胞体系相比，微生物细胞的固定化技术具有以下优点，例如：(i)发酵时间缩短了多至50-60％；(ii)提高了微生物代谢和稳定性；(iii)降低了污染的风险；(iv)细胞密度更高；(v)例如与由于益生菌作用引起后酸化(postacidification)之风险降低相关的稳定的产品品质；(vi)改进底物利用以及(vii)由于持续的细胞再生所致的长时间细胞再利用。
获得这些产品的过程显示了用于在以下多个水平改善发酵过程条件的方法，例如(i)固定化细胞的连续再利用；(ii)发酵时间缩短有助于在整个过程中节能，(iii)降低污染的风险以及(iv)长期维持微生物稳定性。
发明详述
1.燕麦制品混悬液的制备方法
1.1.由薄片、糠和/或粉以及随后在水中混合来制备5-20％(重量/重量)的燕麦制品浓缩物；
1.2.连续搅拌下在80-110℃的温度范围内加热该混合物5-20分钟；
1.3.磨碎所得制备物；
1.4.过滤所得的混悬液；以及
1.5.冷却混合物至25-48℃的温度范围。
2.在流化床反应器中与通过乳化进行细胞包囊相关的预发酵过程
2.1.预发酵过程
利用步骤2.1.1至2.4.3中所得细胞的固定化微生物在流化床反应器中进行预发酵过程。
将囊状物引入孔隙小于所述囊状物直径的柱中诱导微生物-基质在具有恒定和受控的双向氮气流之加压反应器中相互作用。在发酵过程中柱内的固定化细胞得以再次利用直到它们丧失其代谢性质。
2.1.1.细胞培养物的制备
2.1.1.1.由冷冻的培养物制备接种物，随后在补充有0.05％(重量/体积)L-半胱氨酸-HCl的MRS发酵液中连续转移两次进行活化。
2.1.1.2.在1000mL补充有0.05％(重量/体积)L-半胱氨酸-HCl的MRS发酵液(Man Rogosa and Sharpe)中接种1-20％(体积/体积)，例如，对于嗜酸乳杆菌(Lactobacillus acidophilus)Ki而言，随后于厌氧条件下在37℃孵育24小时；对于动物双歧杆菌(Bifidobacteriumanimalis)Bo和Bb12而言，随后于厌氧条件下在37℃孵育48小时。
2.1.1.3.在4℃，以4000rpm离心所得培养物15分钟，随后用例如0.9％(重量/体积)NaCl溶液清洗沉淀物(pellet)，并重悬在100mL同样的溶液中。
2.2.聚合物溶液的制备
2.2.1.聚合物溶液的制备，例如在60-80℃温度下连续搅拌(搅拌时间可从1小时至4小时)1-5％(重量/体积)κ-角叉藻聚糖，之后冷却至35-45℃的温度范围内。
2.3.油溶液的制备
2.3.1.植物油与下述化合物之一的混合物：0.2％(体积/体积)吐温80和/或保护剂(非限定性实例是0.5％(体积/体积)十二烷基硫酸钠)。
2.4.囊状物的制备
2.4.1.1-20％(体积/体积)细胞混悬液(参见2.1)与1-5％(重量/体积)聚合物溶液(参见2.2)的混合物。
2.4.2.将所得混合物加入到75-98％的所制备油溶液(参见2.3.)中。对所得溶液进行均质以形成油包水型乳液。
2.4.3.例如在4-8℃的温度范围内将10mM KCl溶液加入到所述混合物中之后开始形成囊状物。
2.5.预发酵过程操作条件
得到用于流化床反应器的包含微生物的囊状物后，在无菌和厌氧条件(循环氮气流)以及20℃-52℃的温度下进行预发酵过程4-8小时，得到发酵基质。将该混悬液排入反应器中，在该反应器中掺入其余的食品成分。
3.通过乳化和/或喷雾干燥进行的微生物包囊过程
利用下述包囊技术进行微生物的包囊：
3.1.如第2节所述进行乳化；
3.2.喷雾干燥：
3.2.1制备含有聚合物的细胞混悬液(参见2.2、2.2.1以及2.4.1)；
3.2.2.在进口温度和出口温度分别为150-175℃和50-85℃的恒定条件下干燥250ml前述混合物；
3.2.3.将所得粉末以2-5％(重量/体积)的比例以及确保每100g或100mL基质中含有108-1010CFU的方式加入到预发酵的燕麦混悬液(见2.5)中。
4.食品成分的掺入：
将成分以1-3％的浓度范围加入到前述过程(3.2.3)所得基质中，所述食品成分的一个实例是菊粉，非限定性的实例是海盐。
5.基质的呈现形式
基于燕麦混悬液的用包囊益生菌预发酵的共生基质可以新鲜形式、冻干形式和/或冷冻形式呈现。所述新鲜基质还可以凝胶或挤出形式呈现。
Claims (25)
1.一种预发酵共生基质，其特征在于用固定化微生物预发酵谷物混悬液，随后向其加入：
(a)包囊微生物，以及
(b)游离的和/或包囊的益生元，和
(c)其它食品成分。
2.根据权利要求1的预发酵共生基质，其特征在于所述谷物为薄片、粉或糠。
3.根据权利要求2的预发酵共生基质，其特征在于所述谷物是燕麦。
4.根据权利要求1或2的预发酵共生基质，其特征在于所述谷物是通常应用于食品工业中的多于一种谷物的组合，其中所述组合包含或不包含豆类。
5.根据权利要求1或2的预发酵共生基质，其特征在于所述谷物为燕麦与大麦和/或大豆的组合。
6.根据权利要求1的预发酵共生基质，其特征在于所述益生元包含生物活性量的β-葡聚糖可溶性纤维。
7.根据权利要求6的预发酵共生基质，其特征在于所述β-葡聚糖可溶性纤维从谷物提取。
8.根据权利要求6的预发酵共生基质，其特征在于所述β-葡聚糖可溶性纤维从豆类提取。
9.根据权利要求7或8的预发酵共生基质，其特征在于最少含有以重量计0.75％的β-葡聚糖可溶性纤维。
10.根据权利要求6至8中任一项的预发酵共生基质，其特征在于所述益生元还包含其它益生元化合物。
11.根据权利要求10的预发酵共生基质，其中所述其它益生元化合物包括菊粉、果寡糖(FOS)和壳聚糖。
12.根据权利要求1至3和6至8中任一项的预发酵共生基质，其特征在于整合了除益生元功能之外还具有其它功能的其它食品成分。
13.根据权利要求1至3和6至8中任一项的预发酵共生基质，其特征在于整合了除益生元功能之外还具有其它功能的其它食品成分，所述其它功能是感官功能和质构功能。
14.根据权利要求13的预发酵共生基质，其中所述具有感官功能的食品成分是调味剂和/或果浆。
15.根据权利要求13的预发酵共生基质，其中所述具有质构功能的食品成分是酶。
16.根据权利要求1至3和6至8中任一项的预发酵共生基质，其特征在于除了已存在于谷物中的以外，其还包含游离或包囊形式的额外的抗氧化剂来源、脂肪酸、维生素和矿物质。
17.根据权利要求4的预发酵共生基质，其特征在于除了已存在于所述谷物的组合以外，其还包含游离或包囊形式的额外的抗氧化剂来源、脂肪酸、维生素和矿物质。
18.根据权利要求16的预发酵共生基质，其中所述脂肪酸是ω3脂肪酸、ω6脂肪酸。
19.根据权利要求1的预发酵共生基质，其特征在于所述微生物是“GRAS”的益生菌和/或非益生菌，其中在被包囊的情形下，包被材料是下述物质中的至少一种：蛋白质、多糖、脂质和水胶体。
20.根据权利要求19的预发酵共生基质，其特征在于所述微生物属于双歧杆菌属和乳杆菌属。
21.根据权利要求19或20的预发酵共生基质，其特征在于以不低于108CFU/g的量接种微生物，确保当消费时最终产品含有106-108CFU/g。
22.根据权利要求1-3、6-8和19-20中任一项的预发酵共生基质，其特征在于采用新鲜、冻干和/或冷冻形式。
23.根据权利要求22的预发酵共生基质，其特征在于所述新鲜基质采用凝胶或挤出形式。
24.获得根据前述权利要求中任一项的预发酵共生基质的方法，其特征在于通过下述步骤实现：
a)预发酵步骤，将谷物混悬液置于反应器中，所述反应器中含有用蛋白质、多糖、脂质和水胶体包被的大囊状物中的固定化微生物细胞；
b)分离步骤，其中从所述基质中分离出所述微生物；
c)包囊步骤，其中对微生物包囊，或者对微生物和益生元包囊；
d)将所述包囊微生物以及益生元加入谷物混悬液中，其中所述益生元是游离的和/或包囊的；
e)掺入食品成分。
25.权利要求24的方法，其中所述包囊步骤通过干燥、雾化、乳化或凝聚来实现。
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| PCT/PT2007/000042 WO2008041876A2 (en) | 2006-10-06 | 2007-10-04 | Pre-fermented symbiotic matrix based on a cereal suspension with encapsulated probiotics, manufacture process and corresponding uti lization |
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| ES2368401B2 (es) | 2011-09-30 | 2013-06-27 | Universidad Miguel Hernández De Elche | Productos gelificados probióticos o simbióticos y procedimiento para su obtención. |
| CN103013970A (zh) * | 2012-11-29 | 2013-04-03 | 上海致一食品有限公司 | 同时富含植物纤维与益生菌的固定化益生菌及其制备方法 |
| CN104721652A (zh) * | 2015-02-11 | 2015-06-24 | 江苏大学 | 乳酸菌发酵大麦粉及其制备方法与用途 |
| CN107920575A (zh) * | 2015-07-01 | 2018-04-17 | 裕得生物科技有限公司 | 一种合生素发酵产物的制造方法及由此制造的合生素发酵产物 |
| GR1010362B (el) * | 2021-07-06 | 2022-12-21 | Πανας Π. - Κλαουδατος Α. Οε, | Διεργασια ακινητοποιησης προβιοτικων κυτταρων σε βρωμη για παραγωγη προϊοντων-σνακ εμπλουτισμενων με προβιοτικα |
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