CN112890202A - 一种益生菌微囊及其制备方法 - Google Patents

一种益生菌微囊及其制备方法 Download PDF

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CN112890202A
CN112890202A CN202110138512.4A CN202110138512A CN112890202A CN 112890202 A CN112890202 A CN 112890202A CN 202110138512 A CN202110138512 A CN 202110138512A CN 112890202 A CN112890202 A CN 112890202A
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probiotic
preparation
coating
microcapsule
powder
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姚铭飞
韩圣逸
金欣
黄卫新
谢娇娇
陆艳蒙
王波娜
高岭
俞赤慧
李兰娟
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Shaoxing Tongchuang Biotechnology Co ltd
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Shaoxing Tongchuang Biotechnology Co ltd
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Priority to CN202110138512.4A priority Critical patent/CN112890202A/zh
Priority to PCT/CN2021/097408 priority patent/WO2022160540A1/zh
Priority to US17/762,078 priority patent/US20240108047A1/en
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Abstract

本发明提供了一种益生菌微囊及其制备方法，涉及益生菌制品技术领域。所述方法包括以下步骤：(a)载有益生菌的囊芯的制备：将包括益生菌菌粉、微晶纤维素和淀粉在内的囊芯材料混合后，再加入羟丙基甲基纤维素溶液，混合均匀；将得到的混合物料采用挤出滚圆法制备成球状颗粒囊芯；(b)雾化包衣：采用雾化方式将囊芯用包衣材料溶液进行单层或多层包衣，制得核‑壳型微囊。本发明所制备的益生菌微囊，包载量大，微囊颗粒均一，颗粒大小可控；耐贮存，并靶向肠道，耐胃酸且具有高温稳定性。

Description

一种益生菌微囊及其制备方法
技术领域
本发明涉及益生菌制品技术领域，尤其是涉及一种益生菌微囊及其制备方法。
背景技术
益生菌是对肠道具有有益功效的微生物，《益生菌的科学共识》指出:“足够数量、活菌状态和有益健康功能为益生菌的核心特征”。“活菌”是益生菌发挥功效的先决条件，但是益生菌对环境条件非常敏感，不仅菌体极易被氧化，而且被摄入后经过含胃酸、胆盐等复杂上消化道环境还极易失活。
采用微胶囊装包益生菌是当前国内外提升活菌存活率的最有效方法之一。当前的多种制备工艺都存在一定的局限性，例如挤压法效率低，并且微囊粒径大；乳化法需要使用大量油，且分离困难，步骤繁琐；喷雾干燥法则会造成活菌的大量死亡，都不适合用于扩大生产。
因此，特提出本发明。
发明内容
本发明的目的在于提供一种益生菌微囊及其制备方法。本发明所制备的益生菌微囊，包载量大，微囊颗粒均一，颗粒大小可控；耐贮存，并靶向肠道，耐胃酸且具有高温稳定性。
本发明提供的技术方案如下：
一种益生菌微囊的制备方法，所述方法包括以下步骤：
(a)载有益生菌的囊芯的制备：
将包括益生菌菌粉、微晶纤维素和淀粉的囊芯材料混合后，再加入羟丙基甲基纤维素溶液，混合均匀；将得到的混合物料采用挤出滚圆法制备成球状颗粒囊芯；
(b)雾化包衣：采用雾化方式将微囊芯用包衣材料溶液进行单层或多层包衣，制得核-壳型微囊。
在本发明的上下文中，所述的益生菌菌粉，指的是益生菌的冻干粉，包括第一代益生菌和第二代益生菌批准使用的所有菌种。包括例如但不限于：植物乳杆菌、动物双歧杆菌、干酪乳杆菌、副干酪乳杆菌、乳双歧杆菌、短双歧杆菌、鼠李糖乳杆菌、嗜酸乳杆菌、植发酵乳杆菌、肠膜明串珠菌、唾液乳杆菌、瑞士乳杆菌、罗伊氏乳杆菌、格氏乳杆菌、卷曲乳杆菌、约氏乳杆菌、保加利亚乳杆菌、嗜热链球菌、长双歧杆菌、短双歧杆菌、婴儿双歧杆菌、两歧双歧杆菌以及青春双歧杆菌等。
在一个实施方案中，所述囊芯材料还包括脱脂奶粉。
在一个实施方案中，步骤(a)中，所述益生菌菌粉、微晶纤维素、淀粉的质量比为：1～5：40～500：10～250。
优选的，淀粉的种类包括多孔淀粉或抗性淀粉。
在一个实施方案中，所述益生菌菌粉、微晶纤维素、淀粉、脱脂奶粉的质量比为：1～5：40～475：10～250：0～25。
在一个实施方案中，所述益生菌菌粉与所述羟丙基甲基纤维素的质量比为：1～5：0.5～2.5。
在一个实施方案中，所述挤出滚圆法包括以下步骤：
先将混合物料以10～100rpm的速度挤出，并以1500～2000rpm的转速滚圆成球状颗粒。
在一个实施方案中，所述羟丙基甲基纤维素溶液的浓度为0.1～0.5g/mL；优选0.2～0.4g/mL。
在一个实施方案中，雾化包衣的条件为：雾化压力0.1-0.5mpa，优选0.2mpa；入风口的风速为20～25m3/h，优选22～23m3/h，出风口速度为20～25m3/h，优选22～23m3/h；温度为23-27℃，优选25℃；所述包衣材料溶液的流速设置为0.3mL/min～0.6mL/min，优选为0.5mL/min。在一个实施方案中，雾化包衣的时间为：6～10min。
在一个实施方案中，雾化包衣后还包括将包衣好的益生菌微囊冷冻干燥的步骤。
在一个实施方案中，所制备的微囊囊芯的直径在300μm～1.2mm之间，核-壳型微囊直径在300μm～2mm之间。
在一个实施方案中，所述包衣材料可为羟丙基甲基纤维素(HPMC)、明胶、果胶、壳聚糖、黄原胶、阿拉伯胶、抗性淀粉、蛋白粉、聚氯乙烯、醋酸纤维素钛酸酯、羟丙甲纤维素邻苯二甲酸酯或聚乙烯醇钛酸酯中的一种或多种；优选壳聚糖。
在一个实施方案中，所述包衣材料溶液为将包衣材料溶于所述羟丙基甲基纤维素溶液中获得；优选地，所述包衣材料溶液的质量浓度范围为1～10％。
在一个实施方案中，所述制备方法在步骤(a)之前，还包括制备益生菌菌粉的步骤；优选地，通过冷冻干燥法制备菌种冻干粉；更优选地，所述菌粉的活菌数大于1011CFU/g。
本发明还提供了一种益生菌微囊，所述益生菌微囊由上述制备方法制备获得。
本发明还提供了所述益生菌微囊在制备益生菌食品或保健品中的应用。
有益效果：
本发明提供的益生菌微囊制备方法简便，制备步骤可操控性强，用时短，批次之间质量稳定，几乎无差异。
本发明所制备的益生菌微囊，包载量大，微囊颗粒均一，颗粒大小可控，耐贮存，并靶向肠道，耐胃酸且具有高温稳定性，适用于益生菌作为保健品或药品的工业化生产。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案，下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图是本发明的一些实施方式，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为本发明实施例1提供的唾液乳杆菌Li01微囊的形态结构(其中A图为包衣前的微囊囊芯，B图为包衣后的微囊)；
图2为本发明实施例1提供的唾液乳杆菌Li01微囊的抗逆性测定结果(其中A为模拟胃酸环境下进行的测定，B图为模拟肠液环境下进行的测定)；
图3为本发明实施例2提供的唾液乳杆菌Li05微囊的形态结构(其中A图为包衣前的微囊囊芯，B图为包衣后的微囊)；
图4为本发明实施例2提供的Li05微囊的抗逆性测定结果(其中A为模拟胃酸环境下进行的测定，B图为模拟肠液环境下进行的测定)。
具体实施方式
下面将结合实施例对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
实施例1
1.唾液乳杆菌Li01微囊的制备：
(1)乳酸杆菌冻干粉的制备：采用现有的技术，借助发酵罐培养乳杆菌，并进行冷冻干燥，得到唾液乳杆菌Li01冻干粉，其中Li01活菌数大于1011CFU/g。
(2)益生菌微囊囊材的配置：称取Li01益生菌冻干粉1g，微晶纤维素40g和淀粉10g混合制得材料A。
称取HPMC 1g加入50mL 60℃以上的温水混合均匀，制成浓度0.2g/mL溶液B，待B降至室温。将混合好的粉末A倒入托盘中，分多次倒入B溶液，共加入25mL。混合均匀。该步骤和以下的制备过程，均需在洁净室(洁净度为万级)中完成。
(3)用挤出滚圆法制备囊芯：使用孔径为1mm的挤出板，将混合好的物料以36rpm的速度挤出，并用1800rpm的转速滚圆成球状颗粒。
(4)包衣液的制备：将4g壳聚糖溶于B液，制备成包衣液体。
(5)微囊的包衣：取50g左右的囊芯进行包衣：
将入风口速度调至22～23m3/h，温度为25℃，包衣液的流速设置为0.5mL/min,包衣时间为6～10min。
将包衣好的唾液乳杆菌Li01微囊冷冻干燥制成成品。
2.唾液乳杆菌Li01微囊的特征和评价
(1)形态结构：所制备得到的囊芯直接为1mm左右，大小均一；制备的乳酸杆菌微囊在包衣前后尺寸大小变化少。
图1为所提供的唾液乳杆菌Li01微囊的形态结构(其中a图为包衣前的微囊囊芯，B图为包衣后的微囊)。粒径改变较小。
(2)益生菌的包封率：
将微囊轻轻碾碎，配置成混悬液，采用平板计数法计数，测定益生菌的包封率，经测定统计乳酸杆菌微囊的包封率为大于97％。
(3)益生菌的抗逆性：
A、测定益生菌(乳酸杆菌)在模拟胃酸(pH 2.0)、模拟肠液(pH 6.5)中的存活率。
图2为本发明实施例1提供的唾液乳杆菌Li01微囊的抗逆性测定结果(其中A为模拟胃酸环境下进行的测定，B图为模拟肠液环境下进行的测定)。
结果表明：
与未经微囊包埋的乳唾液乳杆菌Li01(Free Li01)相比，微囊包埋的乳唾液乳杆菌Li01(Li01微囊)在胃酸和肠液环境下存活率高。
B、高温(63℃)试验存活率
Figure BDA0002927737530000061
结果表明：
与未经微囊包埋的乳唾液乳杆菌Li01(Free Li01)相比，微囊包埋的乳唾液乳杆菌Li01(Li01微囊)在高温环境下存活率高，且在高温10-40min内任由较高的存活率。
其他性能
本发明实施例1所制备的Li01微囊，将100mg用生理盐水溶解灌胃后，可以在14小时内定植于肠道。
实施例2
1.唾液乳杆菌Li05微囊的制备：
(1)唾液乳杆菌冻干粉的制备：采用现有的技术，借助发酵罐培养唾液乳杆菌Li05，并进行冷冻干燥，得到100g冻干粉，其中戊糖片球菌Li05活菌数大于1011CFU/g。
(2)益生菌微囊囊材的配置：称取Li05益生菌冻干粉2g，微晶纤维素80g和淀粉20g混合制得材料A。
称取HPMC 1g加入50mL 60℃以上的温水混合均匀，制成浓度2g/mL溶液B，待B降至室温。将混合好的粉末A倒入托盘中，分多次倒入B溶液，共加入52mL。混合均匀。该步骤和以下的制备过程，均需在洁净室(洁净度为万级)中完成。
(3)用挤出滚圆法制备囊芯：使用孔径为1mm的挤出板，将混合好的物料以40rpm的速度挤出，并用1700rpm的转速滚圆成球状颗粒。
(4)包衣液的制备：将2g壳聚糖溶于B液，制备成包衣液体。
(5)微囊的包衣：取50g左右的囊芯进行包衣：
将入风口速度调至22m3/h，温度为25℃，包衣液的流速设置为0.4mL/min，包衣时间为8min。
将包衣好的Li05微囊冷冻干燥制成成品。
2.Li05微囊的特征和评价
(1)形态结构：所制备得到的囊芯直接为1mm左右，大小均一；制备的微囊在包衣前后尺寸大小变化少。
图3为所提供的唾液乳杆菌Li05微囊的形态结构(其中a图为包衣前的微囊囊芯，B图为包衣后的微囊)。粒径改变较小。
(2)益生菌的包封率：
将微囊轻轻碾碎，配置成混悬液，采用平板计数法计数，测定益生菌的包封率，经测定统计乳酸杆菌微囊的包封率为大于99％。
(3)益生菌的抗逆性：
A、测定益生菌(乳酸杆菌)在模拟胃酸(pH 2.0)、模拟肠液(pH 6.5)中的存活率。
图4为本发明实施例2提供的Li05微囊的抗逆性测定结果(其中A为模拟胃酸环境下进行的测定，B图为模拟肠液环境下进行的测定)。
结果表明：Li05微囊相比于游离Li05，稳定性显著提高，耐酸和耐胆盐的能力增强。与市面上的微囊相比，该方法制备的微囊颗粒圆，包封率高，大小均一，在包埋过程中益生菌几乎没有损失。
最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。

Claims (10)

1.一种益生菌微囊的制备方法，其特征在于，所述方法包括以下步骤：
(a)载有益生菌的囊芯的制备：
将包括益生菌菌粉、微晶纤维素和淀粉在内的囊芯材料混合后，再加入羟丙基甲基纤维素溶液，混合均匀；将得到的混合物料采用挤出滚圆法制备成球状颗粒囊芯；
(b)雾化包衣：采用雾化方式将囊芯用包衣材料溶液进行单层或多层包衣，制得核-壳型微囊。
2.根据权利要求1所述的制备方法，其特征在于，步骤(a)中，所述挤出滚圆法包括以下步骤：
先将混合物料以10～100rpm的速度挤出，并以1500～2000rpm的转速滚圆成球状颗粒。
3.根据权利要求1所述的制备方法，其特征在于，所述囊芯材料还包括脱脂奶粉。
4.根据权利要求1或3所述的制备方法，其特征在于，步骤(a)中，所述益生菌菌粉、微晶纤维素、淀粉、脱脂奶粉的质量比为：1～5：40～500：20～250：10～25。
5.根据权利要求1所述的制备方法，其特征在于，步骤(a)中，所述益生菌菌粉与所述羟丙基甲基纤维素的质量比为：1～5：0.5～2.5；所述羟丙基甲基纤维素溶液的浓度为0.1～0.5g/mL；优选0.2～0.4g/mL。
6.根据权利要求1所述的制备方法，其特征在于，步骤(b)中，所述雾化包衣的条件为：雾化压力0.1-0.5mpa，优选0.2mpa；入风口的风速为20～25m3/h，优选22～23m3/h，出风口速度为20～25m3/h，优选22～23m3/h；温度为23-27℃，优选25℃；所述包衣材料溶液的流速设置为0.3mL/min～0.6mL/min，优选为0.5mL/min。
7.根据权利要求1所述的制备方法，其特征在于，所述包衣材料溶液为将包衣材料溶于所述羟丙基甲基纤维素溶液中获得；优选地，所述包衣材料溶液的质量浓度范围为1％～10％。
8.根据权利要求1或7所述的制备方法，其特征在于，所述包衣材料为羟丙基甲基纤维素、明胶、果胶、壳聚糖、黄原胶、阿拉伯胶、抗性淀粉、蛋白粉、聚氯乙烯、醋酸纤维素钛酸酯、羟丙甲纤维素邻苯二甲酸酯或聚乙烯醇钛酸酯中的一种或多种；优选壳聚糖。
9.根据权利要求1所述的制备方法，其特征在于，在步骤(a)之前，还包括制备益生菌菌粉的步骤；
优选地，通过冷冻干燥法制备菌种冻干粉；更优选地，制备的菌粉的活菌数大于1011CFU/g。
10.由权利要求1-9任一项所述的制备方法制备获得的益生菌微囊。
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