CN110771898A - 一种益生菌微胶囊、其制备方法和用途 - Google Patents
一种益生菌微胶囊、其制备方法和用途 Download PDFInfo
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Abstract
本发明公开了一种益生菌微胶囊、其制备方法和用途,该益生菌微胶囊的颗粒直径为200‑400μm;在20‑30℃的室温环境下,包埋存活率≥95%;在人工胃液环境里的耐受率≥96%;在人工肠液环境里的缓释时间为120min以上。本发明还公开了该益生菌微胶囊的制备方法。本发明的益生菌微胶囊包埋存活率,便于常温存储和运输,适于作为食品添加剂、保健品添加剂、药品添加剂或饲料添加剂。
Description
技术领域
本发明涉及微生物添加剂领域,具体涉及一种益生菌微胶囊、其制备方法和用途。
背景技术
微生态学研究表明,以双歧杆菌为代表的“益生菌”,其对机体有重要的生理功能,包括抑制肠道致病菌的生长、降低血清中胆固醇含量、激活免疫系统及抗癌活性等作用,随着国内外人们对“益生菌”的认识不断深入,越来越多的益生菌制品在医疗、保健及营养方面得到推广应用。
但是,益生菌制品主要是活菌制剂,大多属专性厌氧菌,极易受到温度、氧气、水分及酸碱等外界环境因素的影响,通常情况活性保持较困难。目前市场上销售的益生菌产品,为保证其在有效期内的活菌数,一般采用2~8℃低温储存,但低温储存条件却限制了产品的储运及市场销售。除此之外,益生菌制品最大的问题是益生菌产品在口服时,容易受到胃酸的作用,在到达起效部位肠道之前活菌数会大幅度下降,从而影响益生菌产品的疗效或保健作用。
为了克服上述问题,在药物和食品工业领域提出了很多益生菌微胶囊化的研究方法。例如:
欧洲专利EP0634167A1公开了胶囊生产工艺,描述了采用三层挤压法:以室温下不流动的疏水物质作为芯材,双歧杆菌菌粉悬浮在芯材中,壁材为①熔点高于体温的油脂,②一定比例的明胶和果胶。芯材和壁材分别进入一个三层喷嘴的内、中、外层,然后同时滴入冷却油中形成直径2.5mm的双歧杆菌胶囊。
日本专利JP57-33543公开了一种保存双歧杆菌的工艺。描述了采用熔点在35℃,温度低于35℃固化的油脂,将双歧杆菌菌粉悬浮于液态油脂中,再降温使油脂固化,包覆住双歧杆菌的工艺。
中国专利CN1310016A公开了另一种生产工艺,利用双歧杆菌的活菌微粒,囊衣液是复合肠溶性材料,通过流化喷雾法制得活性双歧杆菌耐酸肠溶微囊。
中国专利文献CN1613455A公开了一种靶向益生菌微胶囊的制备方法,其包括下列步骤:(1)采用转谷氨酰胺酶交联后的蛋白质将益生菌和/保护剂包埋,并采用冷冻干燥制成初级微胶囊;(2)将初级微胶囊与适当的稀释剂混合后,在流化包衣机中用熔点在30-40℃的氢化油脂包覆制成次级微胶囊;(3)将次级微胶囊用控释包衣材料进行包囊,获得最终微胶囊制品。
上述这些方法的原理都是通过某些物质包覆益生菌,使之与外界空气中的氧、水分隔离,减少外界环境因素对其影响,从而提高益生菌在常温及酸碱条件下稳定性。但EP0634167A1所公开的方法无法避免水的影响(明胶和果胶工艺含水),且去除冷却油的工艺复杂,易影响益生菌存活;JP57-33543所公开的方法工艺条件苛刻,难以直接应用于生产;CN1310016A所公开的方法由于直接采用流化喷雾法,在包囊过程中菌体易造成伤害,从而影响益生菌的稳定性。因此,在益生菌微胶囊方面的方法均存在一定缺陷。另一方面,国内外正在通过生物技术开发高酸、高温耐受性的益生菌变异菌株,通过基因工程构造新菌株来提高益生菌稳定性的方法,但是这种方法需要大量的时间和较高的成本,同时还有可能的安全性争议,目前还没有产品得到应用。而CN1613455A并没有研究制备出的靶向益生菌微胶囊究竟有怎样的性能特性。
发明内容
有鉴于此,为解决上述技术问题,本发明的目的在于提出一种益生菌微胶囊、其制备方法和用途。本发明的益生菌微胶囊具有突出的性能特点。
所采用的技术方案为:
本发明的一种益生菌微胶囊,该益生菌微胶囊的颗粒直径为200-400μm;在20-30℃的室温环境下,包埋存活率≥95%;在人工胃液环境里的耐受率≥96%;在人工肠液环境里的缓释时间为120min以上。
进一步地,该益生菌微胶囊包括益生菌和胶囊壁,所述的胶囊壁包括第一保护层、第二保护层、第三保护层和第四保护层,第一层保护层是用酪蛋白和卵磷脂包埋益生菌形成初级微胶囊;第二层保护层是用淀粉和卵磷脂包覆初级微胶囊形成次级微胶囊;第三保护层是用海藻酸钠、氯化钙和卵磷脂交联包裹次级微胶囊形成耐酸的三级微胶囊,第四保护层用壳聚糖、氯化钙和卵磷脂交联包覆三级微胶囊形成最终的益生菌微胶囊。
进一步地,所述益生菌为选自乳杆菌属、双歧杆菌属中的一种或多种菌株。
本发明的益生菌微胶囊的制备方法,是在流化包衣机中依次完成如下四次包埋步骤:
S1.采用酪蛋白和卵磷脂将益生菌包埋,制成初级微胶囊;
S2.将初级微胶囊用淀粉和卵磷脂包覆制成次级微胶囊;
S3.将次级微胶囊用海藻酸钠、氯化钙和卵磷脂进行包囊,制成三级微胶囊制品;
S4.将三级微胶囊用壳聚糖、氯化钙和卵磷脂进行包覆,制得益生菌微胶囊。
进一步地,在流化包衣机中完成过程的温度为20-30℃。
本发明的益生菌微胶囊的用途中,包括如下:
本发明的益生菌微胶囊用于用于制备食品添加剂。
本发明的益生菌微胶囊用于制备保健品添加剂。
本发明的益生菌微胶囊用于制备饲料添加剂。
本发明的益生菌微胶囊用于制备药物添加剂。
在上述的技术方案中,目前日本的技术所能达到的粒径(颗粒直径)一般能达到0.5mm,但是本发明能够达到粒径为200-400μm。目前的胶囊壁一般为2层或3层,没有发现有采用本发明的四层结构。在20-30℃的室温环境下,包埋存活率≥95%,可以延长益生菌在室温条件下的存活时间,也方便室温下的存储和运输。在人工胃液环境里的耐受率≥96%,可以提高益生菌微胶囊的过胃率;在人工肠液环境里的缓释时间为60min以上;可以使益生菌安全过胃到达肠道,在肠道发挥疗效并提高缓释时间。
在本发明的四层保护层中,每一层都有卵磷脂,能够大大提高相邻各层之间的相容性和粘结力。也是目前没有发现的新技术。
酪蛋白具有较高的表面活性,能够与卵磷脂相容在流化包衣机中完成对益生菌的包埋,而且用酪蛋白包被的益生菌能够提高肠溶的缓释时间;淀粉是由D-葡萄糖通过糖苷键连接而成,用淀粉包埋益生菌提高胃酸的过胃率。海藻酸钠是由D-吡喃甘露糖醛酸和L-吡喃古洛醛酸组成的线性多糖。海藻酸钠能够提高益生菌微胶囊对胃酸的耐受性。壳聚糖[chitosan CS,(1,4)-2-氨基-2-脱氧-B-D-葡聚糖]是一种性质活泼的高分子聚合物,是由甲壳质脱乙酰基后生成的产物。低分子量壳聚糖做成的微胶囊能更好的控制释放速率,提高肠溶缓释时间。
综上,通过本发明制备的益生菌微胶囊具有突出的性能特点,是目前现有技术所不具备的。
具体实施方式
下面通过具体的实施例对本发明进行详细说明,但这些例举性实施方式的用途和目的仅用来例举本发明,并非对本发明的实际保护范围构成任何形式的任何限定,更非将本发明的保护范围局限于此。
实施例1
一种益生菌微胶囊的制备方法,其是在流化包衣机中20-30℃下依次完成如下四次包埋步骤:
S1.采用酪蛋白和卵磷脂将益生菌包埋,制成初级微胶囊;
S2.将初级微胶囊用淀粉和卵磷脂包覆制成次级微胶囊;
S3.将次级微胶囊用海藻酸钠、氯化钙和卵磷脂进行包囊,制成三级微胶囊制品;
S4.将三级微胶囊用壳聚糖、氯化钙和卵磷脂进行包覆,制得益生菌微胶囊。
本实施例具体步骤如下:
S1.初级微胶囊的制备
a.复合包埋液的制备
取100ml去离子水,加入10g酪蛋白,溶混匀搅拌10分钟,在37℃水浴锅中恒温30分钟,制成包埋液。
b.将含有双歧杆菌的发酵液12公斤置于管式(4000-7000rpm)离心机离心收集菌体,每公斤所获湿菌泥加入10ml包埋液,混匀后置于真空冷冻干燥机内,干燥24-36小时,然后置于流化包衣机内(北京长征天民高科技有限公司的BZT360型)喷涂卵磷脂,制得冻干双歧杆菌初级微胶囊。
S2.次级微胶囊制备
a.60g双歧杆菌初级微胶囊与100g淀粉按等倍稀释法混合均匀备用。
b.将上述混合物置于流化包衣机内(北京长征天民高科技有限公司的BZT360型)喷涂卵磷脂进行包埋操作,包覆时间为30-40分钟。按上述条件制得双歧杆菌次级微胶囊。
S3.三级微胶囊制备
a.60g双歧杆菌初级微胶囊、海藻酸钠40g和氯化钙40g,溶于体积浓度80%的乙醇,定容至1000ml。
b.将上述混合物置于流化包衣机内(北京长征天民高科技有限公司的BZT360型)喷涂卵磷脂进行包埋操作,包覆时间为30-40分钟。按上述条件制得双歧杆菌三级微胶囊。
S4.最终的益生菌微胶囊制备
a.60g双歧杆菌初级微胶囊、壳聚糖10g和氯化钙30g,溶于体积浓度80%的乙醇,定容至1000ml。
b.将上述混合物置于流化包衣机内(北京长征天民高科技有限公司的BZT360型)喷涂卵磷脂进行包埋操作,包覆时间为30-40分钟。按上述条件制得最终的双歧杆菌微胶囊制品。
得到的双歧杆菌微胶囊的颗粒直径为200-400μm;在20-30℃的室温环境下,包埋存活率≥95%;在人工胃液环境里的耐受率≥96%;在人工肠液环境里的缓释时间为120min以上。
其中微胶囊粒径的测定是通过用荧光摄影显微镜测定大量微胶囊的粒径。包埋存活率的测定是=微胶囊中活菌数/微胶囊化前的活菌数x100%。
益生菌微胶囊耐酸性的测定:是取微胶囊0.1g置于盛有50mL,pH1.2的人工胃液的三角瓶中,于摇床中以37±1℃,180rpm/min,处理0,l,2,3h时取样测定其透光率,根据透光率变化来分析人工胃液中微胶囊的溶出情况,用透光率的值来间接表达益生菌微胶囊在人工胃液环境里的耐受率。
益生菌微胶囊肠液性测定:将微胶囊0.1g置于盛有50mL,pH6.8人工肠液的三角瓶中,于摇床中以以37±1℃,180rpm/min进行崩解.分别于0,l5,30,45,60,120min时取样测其透光率,根据透光率变化来分析人工肠液中微胶囊的溶出情况。实验表明,在120min后,仍有5.24%的透光率。
该双歧杆菌微胶囊可用于制备食品添加剂、保健品添加剂、饲料添加剂或药物添加剂。
实施例2
参照实施例1,与实施例1不同的是,本实施例用的是乳杆菌,制得的是乳杆菌微胶囊制品。
得到的乳杆菌微胶囊仍具有实施例1的双歧杆菌微胶囊的特性:颗粒直径为200-400μm;在20-30℃的室温环境下,包埋存活率≥95%;在人工胃液环境里的耐受率≥96%;在人工肠液环境里的缓释时间为120min以上。
该乳杆菌微胶囊可用于制备食品添加剂、保健品添加剂、饲料添加剂或药物添加剂。
上文所列出的一系列的详细说明仅仅是针对本发明的可行性实施例的具体说明,它们并非用以限制本发明的保护范围,凡未脱离本发明技艺精神所作的等效实施例或变更均应包含在本发明的保护范围之内。
Claims (9)
1.一种益生菌微胶囊,其特征在于,该益生菌微胶囊的颗粒直径为200-400μm;在20-30℃的室温环境下,包埋存活率≥95%;在人工胃液环境里的耐受率≥96%;在人工肠液环境里的缓释时间为120min以上。
2.根据权利要求1所述的益生菌微胶囊,其特征在于,包括益生菌和胶囊壁,所述的胶囊壁包括第一保护层、第二保护层、第三保护层和第四保护层,第一层保护层是用酪蛋白和卵磷脂包埋益生菌形成初级微胶囊;第二层保护层是用淀粉和卵磷脂包覆初级微胶囊形成次级微胶囊;第三保护层是用海藻酸钠、氯化钙和卵磷脂交联包裹次级微胶囊形成耐酸的三级微胶囊,第四保护层用壳聚糖、氯化钙和卵磷脂交联包覆三级微胶囊形成最终的益生菌微胶囊。
3.根据权利要求2所述的益生菌微胶囊,其特征在于,所述益生菌为选自乳杆菌属、双歧杆菌属中的一种或多种菌株。
4.一种权利要求1-3任一所述的益生菌微胶囊的制备方法,其特征在于,在流化包衣机中依次完成如下四次包埋步骤:
S1.采用酪蛋白和卵磷脂将益生菌包埋,制成初级微胶囊;
S2.将初级微胶囊用淀粉和卵磷脂包覆制成次级微胶囊;
S3.将次级微胶囊用海藻酸钠、氯化钙和卵磷脂进行包囊,制成三级微胶囊制品;
S4.将三级微胶囊用壳聚糖、氯化钙和卵磷脂进行包覆,制得益生菌微胶囊。
5.根据权利要求4所述的益生菌微胶囊的制备方法,其特征在于,在流化包衣机中完成过程的温度为20-30℃。
6.权利要求1-3任一所述的益生菌微胶囊用于制备食品添加剂。
7.权利要求1-3任一所述的益生菌微胶囊用于制备保健品添加剂。
8.权利要求1-3任一所述的益生菌微胶囊用于制备饲料添加剂。
9.权利要求1-3任一所述的益生菌微胶囊用于制备药物添加剂。
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