CN114886121A - 一种肠道益生菌高活性胶珠制备及应用 - Google Patents
一种肠道益生菌高活性胶珠制备及应用 Download PDFInfo
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- CN114886121A CN114886121A CN202210540993.6A CN202210540993A CN114886121A CN 114886121 A CN114886121 A CN 114886121A CN 202210540993 A CN202210540993 A CN 202210540993A CN 114886121 A CN114886121 A CN 114886121A
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- Prior art keywords
- solution
- probiotic
- sodium alginate
- beads
- glue
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Abstract
本发明公开了一种肠道益生菌高活性胶珠制备及应用,属于食品加工技术领域。本发明以海藻酸钠溶液或不同组合混合溶液为固定化细胞载体对益生菌进行固定,不仅提高了益生菌的存活率,同时由于除海藻酸钠以外其他成分的加入,调整了益生菌胶珠的机械强度,提高了益生菌包埋颗粒包埋率和模拟胃肠道环境下的活菌数,益生菌胶珠在胃中稳定性提高,肠道内释放彻底;在研发果汁、茶饮料过程中,通过加入制备的益生菌胶珠,不仅使产品的风味和外观得以改善,同时赋予产品一定的功能特性。
Description
技术领域
本发明涉及一种肠道益生菌高活性胶珠制备及应用,属于食品加工技术领域。
背景技术
乳酸菌(lactic acid bacteria,LAB)是一类能利用可发酵碳水化合物产生大量乳酸的细菌的统称。LAB是一种益生菌,其通过发酵产生的有机酸等物质可调节胃肠道正常菌群、维持微生态平衡、提高机体免疫力等。
据报道,益生菌在食物基质中的生存力应≥106CFU/mL(g),然而,在处理、储存过程中,有些因素会降低益生菌的活性,这些因素包括pH、氧气和温度等。益生菌在通过人体胃肠道时,需要经过胃、小肠等消化器官,里面存在的胃酸、胆汁、消化酶等使它们输送到胃肠道目标部位期间的活力降低,难以高效在肠道定植,所以达不到或益生效果不佳,应用受到限制。因此,提高益生菌对体内环境的耐受程度和存活率是益生菌发挥作用的关键。已有研究提出了提高益生菌在不利条件下的生存能力的方法,包括选择耐酸和耐胆汁的菌株、加入微量营养素、利用益生菌之间的协同作用来增加存活率和提高稳定性等,但目前的应用效果并未得到改善。
为保持益生菌活性、到达肠道内的活菌数量及有效释放这一难题,本专利提供一种改进的气控-包埋方法,通过合适的材料将益生菌包裹起来的同时充分进入无菌空气,这样既可以使益生菌较少的接触胃酸等刺激性物质,又可以将益生菌在肠道内进行有效释放,使其在肠道内高效定植,从而真正高效发挥益生菌的益生功效。
发明内容
本发明以海藻酸钠溶液或不同组合混合溶液为固定化细胞载体对益生菌进行固定,不仅提高了益生菌的存活率,同时由于除海藻酸钠以外其他成分的加入,调整了益生菌胶珠的机械强度,提高了益生菌包埋颗粒包埋率和模拟胃肠道环境下的活菌数,益生菌胶珠在胃中稳定性提高,肠道内释放彻底;在研发果汁、茶饮料过程中,通过加入制备的益生菌胶珠,不仅使产品的风味和外观得以改善,同时赋予产品一定的功能特性。
本发明提供了一种益生菌胶珠,所述益生菌胶珠按照以下方法制备:
(1)将载体溶液与益生菌菌悬液混合后,得到混合溶液;所述载体溶液为:海藻酸钠溶液、淀粉溶液、低聚果糖溶液、乳蔗糖溶液中的一种或多种;
(2)按照0.5~1.2L/min的通气量向混合溶液中持续通入空气后,将混合溶液滴入钙离子溶液中进行固化后制备得到益生菌胶珠。
在本发明的一种实施方式中,步骤(1)中载体溶液中溶质的浓度为10~50g/L;所述益生菌菌悬液与载体溶液添加的体积比为:1:(8~10)。
在本发明的一种实施方式中,步骤(2)为,在0.2~0.4kg/cm2压力条件下,按照0.5~1.2L/min的通气量向混合溶液中持续通入直径为2~5μm无菌空气,通气时间为:0.5~1h;同时,将混合溶液滴入到质量分数为0.2~1.0%的钙离子溶液中,固化30~50min,过滤后制备得到。
在本发明的一种实施方式中,所述益生菌胶珠外壁还涂覆有壳聚糖。
在本发明的一种实施方式中,所述载体溶液为海藻酸钠溶液,所述海藻酸钠溶液的浓度为10~50g/L。
在本发明的一种实施方式中,所述载体溶液为低聚果糖-海藻酸钠溶液,所述低聚果糖-海藻酸钠溶液是将低聚果糖溶液和海藻酸钠溶液按照体积比为(1~1.5):1的比例混合后得到的,其中,所述低聚果糖溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
在本发明的一种实施方式中,所述载体溶液为淀粉-海藻酸钠溶液,所述淀粉-海藻酸钠溶液是将淀粉溶液和海藻酸钠溶液按照体积比为(1~1.5):1混合后得到的,其中,所述淀粉溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
在本发明的一种实施方式中,所述载体溶液为乳蔗糖-海藻酸钠溶液,所述乳蔗糖-海藻酸钠溶液是将乳蔗糖溶液和海藻酸钠溶液按照体积比为(1~1.5):1混合后得到的,其中,所述乳蔗糖溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
在本发明的一种实施方式中,所述益生菌包括但不限于,嗜酸乳杆菌(Lactobacillus acidophilus)、植物乳杆菌(Lactobacillus plantarum)、干酪乳杆菌(Lactobacillus casei)、保加利亚乳杆菌(Lactobacillus bulgaricus)、鼠李糖乳杆菌(Lactobacillus rhamnosus)和嗜热链球菌(Streptococcus thermophilus)中的一种或几种。
在本发明的一种实施方式中,所述益生菌菌悬液制备方法为:将益生菌菌剂进行活化、培养后收集菌体,用0.9%无菌生理盐水制成益生菌菌悬液,活菌数≥1.0×109CFU/mL。
在本发明的一种实施方式中,所述益生菌胶珠的内部均匀分布直径2~5μm的气泡,气泡体积占胶珠总体积的5~12%,通过氧气控制了益生菌的代谢,延长了益生菌活性,使胶珠保藏期延长了15d。
在本发明的一种实施方式中,所述益生菌胶珠具有良好的稳定性,在4℃条件储藏30d后,含有空气的益生菌胶珠活菌数仍保持在108CFU/g及以上。
在本发明的一种实施方式中,所述益生菌胶珠在模拟胃消化阶段(pH 2.0),2h内活菌未发生明显释放,且活细胞仍保持在71%以上;在模拟肠消化阶段(pH 7.4),0.5h内胶珠彻底破裂,活细胞得以充分释放,释放率为90%以上。
在本发明的一种实施方式中,所述益生菌胶珠在pH 1.5~5.0溶液条件下放置4h后,刚性表现为由4500~6500g降低至2500~2200g;
在pH 5.5~7.5溶液条件下放置0.1~4h后,胶珠刚性表现为1800~1000g,且极易在肠环境下破裂,这对其在肠消化阶段释放活细胞起到积极作用。
本发明还提供了一种含有上述益生菌胶珠的产品。
在本发明的一种实施方式中,所述产品为食品、药品或保健品。
在本发明的一种实施方式中,所述食品为果汁、茶饮料、奶酪、酸奶。
在本发明的一种实施方式中,所述药品或保健品为胶囊、片剂。
在本发明的一种实施方式中,按饮料与所述益生菌胶珠质量比为(1000-1200):1的比例混合后直接饮用,或暂贮于4℃冰箱中。
本发明还提供了上述益生菌胶珠,或上述产品在提高益生菌在胃肠道中活性中的应用。
本发明还提供了上述益生菌胶珠,或上述产品在制备提高益生菌在胃肠道中活性的产品中的应用。
本发明还提供了一种制备高活性益生菌胶珠的方法,所述方法包括以下步骤:
(1)将载体溶液与益生菌菌悬液混合后,得到混合溶液;所述载体溶液为海藻酸钠溶液、淀粉溶液、低聚果糖溶液、乳蔗糖溶液中的一种或多种;
(2)按照0.5~1.2L/min的通气量向混合溶液中持续通入空气后,将混合溶液滴入钙离子溶液中进行固化后制备得到益生菌胶珠。
在本发明的一种实施方式中,步骤(1)中,所述载体溶液的浓度为10~50g/L;所述益生菌菌悬液与载体溶液添加的体积比为:1:(8~10)。
在本发明的一种实施方式中,步骤(2)中的钙离子溶液的质量分数为0.2~1.0%,进行固化的时间为30~50min。
在本发明的一种实施方式中,所述方法还包括,将所述益生菌胶珠外壁涂覆有壳聚糖。
在本发明的一种实施方式中,所述载体溶液为海藻酸钠溶液,所述海藻酸钠溶液的浓度为10~50g/L;
或所述载体溶液为低聚果糖-海藻酸钠溶液,所述低聚果糖-海藻酸钠溶液是将低聚果糖溶液和海藻酸钠溶液按照体积比为(1~1.5):1的比例混合后得到的,其中,所述低聚果糖溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
或所述载体溶液为淀粉-海藻酸钠溶液,所述淀粉-海藻酸钠溶液是将淀粉溶液和海藻酸钠溶液按照体积比为(1~1.5):1混合后得到的,其中,所述淀粉溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
或所述载体溶液为乳蔗糖-海藻酸钠溶液,所述乳蔗糖-海藻酸钠溶液是将乳蔗糖溶液和海藻酸钠溶液按照体积比为(1~1.5):1混合后得到的,其中,所述乳蔗糖溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
有益效果
(1)益生菌胶珠制备过程中,向载体溶液中通气0.5~1h,使得胶珠内部均匀分布直径2~5μm的气泡,气泡体积占胶珠总体积的5~12%,通过氧气控制了益生菌的代谢,延长了益生菌活性,使胶珠保藏期延长了15d。
(2)采用本发明的方法,相较于游离的益生菌细胞溶液,益生菌胶珠表现出较好的存活力,在模拟胃消化120min后活细胞存活率保持在71%以上,模拟肠消化30min内活菌释放90%以上。
(3)采用本发明的方法,相较于单一的海藻酸钠益生菌胶珠,不同组合包封材料制备的益生菌胶珠的包封率在68.7~79.2%,提高了8.3~11.1%。
(4)采用本发明的方法,相较于游离的益生菌细胞悬液,包埋对益生菌的生长没有影响。
附图说明
图1:胶珠制备示意图。
图2:益生菌胶珠(含空气)的SEM图像,每列图像的放大倍数分别为20×、2400×和5000×,每行图像分别显示;其中,A为海藻酸钠益生菌胶珠;B为低聚果糖-海藻酸钠益生菌胶珠。
图3:不同益生菌胶珠的稳定性检测;其中,A:游离细胞;B:海藻酸钠益生菌胶珠(含空气);C:海藻酸钠-壳聚糖益生菌胶珠(含空气);D:淀粉-海藻酸钠益生菌胶珠(含空气);E:低聚果糖-海藻酸钠益生菌胶珠(含空气);F:海藻酸钠益生菌胶珠(不含空气);G:海藻酸钠-壳聚糖益生菌胶珠(不含空气);H:淀粉-海藻酸钠益生菌胶珠(不含空气);I:低聚果糖-海藻酸钠益生菌胶珠(不含空气)。
图4:模拟胃肠道环境后益生菌活细胞存活率;其中,A:游离细胞;B:海藻酸钠益生菌胶珠(含空气);C:海藻酸钠-壳聚糖益生菌胶珠(含空气);D:淀粉-海藻酸钠益生菌胶珠(含空气);E:低聚果糖-海藻酸钠益生菌胶珠(含空气);F:海藻酸钠益生菌胶珠(不含空气);G:海藻酸钠-壳聚糖益生菌胶珠(不含空气);H:淀粉-海藻酸钠益生菌胶珠(不含空气);I:低聚果糖-海藻酸钠益生菌胶珠(不含空气)。
图5:不同益生菌胶珠对益生菌生长的影响;其中,A:游离细胞;B:海藻酸钠益生菌胶珠(不含空气);C:淀粉-海藻酸钠益生菌胶珠(不含空气);D:低聚果糖-海藻酸钠益生菌胶珠(不含空气)。
具体实施方式
下面通过实施例对本发明进一步说明,实施例只用于对本发明进行进一步的说明,不能理解为对本发明保护范围的限制,本领域的技术人员根据上述本发明的内容做出的一些非本质的改进和调整也属于本发明保护的范围。
本发明中采用的益生菌为Lactobacillus plantarum CICC 25034、Lactobacillus acidophilus CICC 22162、Lactobacillus rhamnosus CICC 20255中的一种或几种,购自中国工业微生物菌种保藏管理中心。
下述实施例中以Lactobacillus plantarum CICC 25034为例,仅仅是为了说明本实验的技术效果,并不能成为本发明的限定。
下述实施例中所涉及的培养基如下:
MRS培养基(g/L,液体):葡萄糖20;蛋白胨10;牛肉浸膏10;酵母浸膏5;乙酸钠2;柠檬酸氢二铵2;K2HPO4 2;MgSO4·7H2O 0.58;MnSO4·H2O 0.25;吐温80 1mL;pH 6.2±0.2。115℃灭菌20min。
MRS培养基(固体):上述液体培养基中加1.5-2%琼脂,115℃灭菌20min。
下述实施例中所涉及的检测方法如下:
包封率的测定
包封率=N/N0·100,其中,N为益生菌胶珠包埋的活细胞数,N0是最初添加活游离细胞总数。
储藏稳定性的检测
将益生菌胶珠在4℃、25℃下分别储藏,浸泡于水中或未浸泡,每隔3d取样,对活细胞数量通过平板计数法进行测定。首先,取1g胶珠于9mL无菌3%柠檬酸钠溶液中,漩涡震荡0.5h,使胶珠完全崩解。然后,取1mL样品悬液,用无菌0.9%生理盐水连续稀释,选取106、107两个稀释度,取100μL接种于MRS琼脂平板上,每个稀释度三个平行。37℃培养箱中培养48h后进行活菌计数。用游离细胞溶液作为对照。
实施例1:海藻酸钠益生菌胶珠制备及包封率测定
具体步骤如下:
(1)益生菌菌悬液及海藻酸钠溶液的制备(如图1所示)
将益生菌接种至MRS培养基中,于37℃培养,取对数生长期(OD值为8.56)培养液离心,收集菌体,菌体用0.9%无菌生理盐水制成益生菌菌悬液,经检测,活菌数为1.0×109CFU/mL;
将海藻酸钠添加至无菌水中,制备得到浓度为20g/L的海藻酸钠溶液。
(2)海藻酸钠益生菌胶珠的制备
将步骤(1)制备得到的益生菌菌悬液和海藻酸钠溶液,按照体积比为1:8的比例,混合,并装入容器中,在0.2kg/cm2压力条件下,向载体溶液中持续通入无菌空气(通气量为:0.5L/min),通气时间为:0.5h,使胶珠内部均匀分布直径为2μm的气泡,同时将溶液逐滴滴入到质量分数为0.2%的CaCl2溶液中,固化40min,过滤后用无菌去离子水洗涤,得到海藻酸钠益生菌胶珠(含空气),其SEM图像如图2所示。
(3)包封率的检测
取1g步骤(2)得到的胶珠于9mL 3%柠檬酸钠溶液中使其破碎,用无菌0.9%生理盐水进行梯度稀释,得到106、107两个稀释度的样品悬液;
分别取100μL上述样品悬液,接种于MRS培养基中,37℃培养48h,按照GB4789.35-2016进行活菌计数。
经测定,其包封率为69.4%。
(4)对照胶珠
具体实施方式同步骤(1)~(2),区别在于,步骤(2)为:将步骤(1)制备得到的益生菌菌悬液和海藻酸钠溶液,按照体积比为1:8的比例,混合,将溶液逐滴滴入到质量分数为0.2%的CaCl2溶液中,固化40min,过滤后用无菌去离子水洗涤,得到海藻酸钠益生菌胶珠(不含空气)。
实施例2:涂覆有壳聚糖的海藻酸钠益生菌胶珠的制备
(1)1%壳聚糖溶液的制备:
按照质量比1:99的比例将乙酸与水充分混合,制成1%乙酸溶液,1g壳聚糖加入100mL1%乙酸溶液中得质量分数为1%的壳聚糖溶液。
(2)取制备好的1%壳聚糖溶液,按照质量比1:10的比例,将实施例1中制备得到的海藻酸钠益生菌胶珠(含空气)加入1%壳聚糖溶液中,100r/min,摇床搅拌10min,过滤,制备得到涂覆有壳聚糖的海藻酸钠益生菌胶珠(含空气)。
(3)将上述胶珠用无菌0.1mol/L的PBS缓冲液冲洗后,于4℃冰箱中贮存备用。
包封率测定方法同实施例1,经测定,其包封率为72.4%。
(4)对照胶珠
将实施例1制备得到的对照胶珠按照步骤(1)~(2)的方法制备得到涂覆有壳聚糖的海藻酸钠益生菌胶珠(不含空气)。
实施例3:海藻酸钠与其他包封材料组合制备益生菌胶珠及包封率测定
1、淀粉-海藻酸钠益生菌胶珠
(1)益生菌菌悬液及海藻酸钠溶液的制备
将益生菌接种至MRS培养基中,于37℃培养,取对数生长期(OD值为8.56)培养液离心,收集菌体,菌体用0.9%无菌生理盐水制成益生菌菌悬液,经检测,活菌数为1.0×109CFU/mL;
将海藻酸钠添加至无菌水中,制备得到20g/L的海藻酸钠溶液,淀粉添加至无菌水中,制备得到20g/L的淀粉溶液,将制备得到的海藻酸钠溶液与淀粉溶液按照体积比为1:1的比例制备得到淀粉-海藻酸钠溶液。
(2)将步骤(1)制备得到的益生菌菌悬液和淀粉-海藻酸钠溶液,按照体积比1:10的比例,将益生菌菌悬液和混合载体溶液装入容器中并充分搅拌,向溶液中通气充入直径为3μm的无菌空气(通气量为:0.7L/min),通气时间为:0.6h,同时,将溶液逐滴滴入到质量分数为0.4%的CaCl2溶液中,固化40min,过滤后用无菌去离子水洗涤,得到淀粉-海藻酸钠益生菌胶珠(含空气)。
包封率测定方法同实施例1,经测定,其包封率为79.2%。
(3)对照胶珠
具体实施方式同步骤(1)~(2),区别在于,步骤(2)为,将步骤(1)制备得到的益生菌菌悬液和淀粉-海藻酸钠溶液,按照体积比1:10的比例,将益生菌菌悬液和混合载体溶液装入容器中并充分搅拌,将溶液逐滴滴入到质量分数为0.4%的CaCl2溶液中,固化40min,过滤后用无菌去离子水洗涤,得到淀粉-海藻酸钠益生菌胶珠(不含空气)。
2、低聚果糖-海藻酸钠益生菌胶珠
(1)益生菌菌悬液及海藻酸钠溶液的制备
将益生菌接种至MRS培养基中,于37℃培养,取对数生长期(OD值为8.56)培养液离心,收集菌体,菌体用0.9%无菌生理盐水制成益生菌菌悬液,经检测,活菌数为1.0×109CFU/mL;
将海藻酸钠添加至无菌水中,制备得到20g/L的海藻酸钠溶液,低聚果糖添加至无菌水中,制备得到20g/L的低聚果糖溶液,将制备得到的海藻酸钠溶液与低聚果糖溶液按照体积比为1:1的比例制备得到低聚果糖-海藻酸钠溶液。
(2)将步骤(1)制备得到的益生菌菌悬液和低聚果糖-海藻酸钠溶液,按照体积比1:12的比例,将益生菌菌悬液和混合载体溶液充分搅拌,并装入容器中,充入直径为4μm的无菌空气(通气量为:1.0L/min),通气时间为:0.8h,同时将溶液逐滴滴入到质量分数0.6%的CaCl2溶液中,固化40min,过滤后用无菌去离子水洗涤,得到低聚果糖-海藻酸钠益生菌胶珠(含空气),其SEM图像如图2所示。
包封率测定方法同实施例1,其包封率为74.6%。
(3)对照胶珠
具体实施方式同步骤(1)~(2),区别在于,步骤(2)为,将步骤(1)制备得到的益生菌菌悬液和低聚果糖-海藻酸钠溶液,按照体积比1:12的比例,将益生菌菌悬液和混合载体溶液充分搅拌,将溶液逐滴滴入到质量分数0.6%的CaCl2溶液中,固化40min,过滤后用无菌去离子水洗涤,得到低聚果糖-海藻酸钠益生菌胶珠(不含空气)。
实施例4:不同益生菌胶珠的稳定性及性能研究
(1)将实施例1-3制备的益生菌胶珠、对照胶珠于4℃下储藏,每隔3d取样,利用平板计数法检测活菌数量,以游离细胞溶液作为对照(如图3所示)。
结果显示,储存30天后,游离细胞溶液的活菌数量明显减少,从109CFU/mL降至105CFU/mL,不含空气的益生菌胶珠活菌数从109CFU/g下降到107CFU/g,而含有空气的益生菌胶珠表现为109CFU/g到108CFU/g的平缓下降。经包埋后,在储存过程中益生菌的生存能力得到了提高,具有优良的稳定性。
(2)使用TA-XT plus物性分析仪对不同益生菌胶珠的弹性进行测定(表1)。
表1不同益生菌胶珠的弹性比较
结果表明,含空气的益生菌胶珠弹性较大,其弹性为26.97-35.64%,而不含空气的益生菌胶珠弹性在16.72-22.74%范围内。
实施例5:不同益生菌胶珠在模拟胃肠环境条件下的生存能力
具体步骤如下:
(1)溶液的配制:
模拟胃液:制备0.2g NaCl,1g胃蛋白酶,蒸馏水100mL,1mol/L HCl调pH至2.0,用0.22μm水系滤菌器过滤除菌。
模拟肠液:1.2g KH2PO4、0.45g胆盐,1g胰蛋白酶,蒸馏水100mL,1mol/L NaOH调pH至7.4,用0.22μm水系滤菌器过滤除菌。
(2)模拟胃环境实验
分别将实施例1-3制备得到的1g益生菌胶珠、对照胶珠,放入含有9ml模拟胃液的试管中,放置于37℃,120r/min的摇床中,在0、1、2h取样,并通过活细胞计数进行分析后计算存活率(图4),模拟胃环境2h后活细胞释放率见表2。
(3)模拟肠道环境实验
将步骤(2)中实验结束后,除去胃溶液,加入5mL无菌模拟肠液中,37℃,100r/min,在0、1、2、3h时取样,并通过活细胞计数进行分析,同时以游离细胞进行对照实验(图4),模拟肠道环境0.5h后活细胞释放率见表2。
表2:不同益生菌胶珠在模拟胃肠环境条件下活细胞释放情况
模拟胃消化实验(pH 2.0)2h后,游离细胞的活细胞数损失了80%,而经包埋的益生菌胶珠的活细胞存活率在71%以上;模拟肠消化实验(pH 7.4)0.5h后,游离细胞未检测到活菌数,经包埋的益生菌胶珠的活细胞释放率在90%以上。
同时,测定了不同益生菌胶珠在模拟胃消化2h、模拟肠消化0.5h后的刚性,结果见表3。
表3:不同益生菌胶珠的刚性变化
结果显示,在经过胃环境2h、肠环境0.5h后,不同益生菌胶珠的刚性发生明显下降,这有利于胶珠在模拟肠消化阶段的活菌释放。
对比例1:海藻酸钠益生菌胶珠制备及包封率测定
具体实施方式同实施例1的步骤(1)~(3),区别在于,调整步骤(2)中充入的无菌空气的直径分别为:2μm(通气量为:0.5L/min)、3μm(通气量为:0.7L/min)、4μm(通气量为:1.0L/min),制备得到海藻酸钠益生菌胶珠(含空气),结果显示,其包封率分别为:78.6%、73.5%、70.2%。
具体实施方式同实施例1的步骤(1)~(3),区别在于,调整步骤(2)中钙离子溶液的浓度分别为0.2%、0.6%、1.0%,制备得到海藻酸钠益生菌胶珠(含空气),结果显示,其包封率分别为:69.2%、79.0%、72.3%。
实施例6:包埋对益生菌生长的影响
益生菌胶珠的制备如实施例1~3所述,制备得到:海藻酸钠益生菌胶珠(不含空气)、淀粉-海藻酸钠益生菌胶珠(不含空气)、低聚果糖-海藻酸钠益生菌胶珠(不含空气)。
分别将1g益生菌胶珠、1mL游离细胞悬液接种于99mL MRS液体培养基中,通过监测它们的生长曲线来比较包埋对益生菌生长的影响。结果如图5所示。
从图中可以看出,四个测试组都表现出典型的相似的微生物生长曲线,这表明包埋后,仍然可以保持益生菌的生长能力。
实施例7:柠檬百香果茉莉益生菌茶饮料
(1)按照实施例3的方法制备得到含空气的淀粉-海藻酸钠益生菌胶珠。
(2)柠檬百香果茉莉益生菌茶饮料制备:将百香果洗净取果肉,柠檬洗净切片,25g百香果果肉于500mL PP杯中,将冲泡好的茉莉花茶(带3-5颗茉莉花)与制备好的淀粉-海藻酸钠益生菌胶珠按质量比1200:1的比例混合,最后加入2片柠檬片,得到柠檬百香果茉莉益生菌茶饮料,直接饮用或暂贮于4℃冰箱中。
(3)柠檬百香果茉莉益生菌茶饮料感官评价:对茶饮料的色泽、风味、口感、组织状态进行评价,感官评价小组由10名食品专业人员组成,最终结果见表4。
表4益生菌茶饮料/果汁感官分析
结果显示,益生菌胶珠的加入赋予益生菌茶饮料/果汁以良好的口感和组织状态,同时又使其具备一定的功能特性。
实施例8:青柠益生菌芒果汁制备
(1)按照实施例3的方法分别制备得到含空气的低聚果糖-海藻酸钠益生菌胶珠。
(2)青柠益生菌芒果汁制备:将芒果洗净,去皮,切成果肉丁,按加水量:果肉重量=1:1的比例加水,榨汁,加0.1%VC防止褐变;青柠洗净,切薄片。将芒果汁与低聚果糖-海藻酸钠益生菌胶珠按质量比1000:1的比例混合并搅拌,最后加入2-3片青柠,得到青柠益生菌芒果汁,直接饮用或暂贮于4℃冰箱中。
(3)青柠芒果汁益生菌饮料感官评价:同实施例6;
(4)青柠芒果汁益生菌饮料性能测定:同实施例6。使用未添加益生菌胶珠的芒果汁进行对照实验,结果如表5所示。
表5益生菌茶饮料/果汁感官分析
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (10)
1.一种益生菌胶珠,其特征在于,所述益生菌胶珠按照以下方法制备:
(1)将载体溶液与益生菌菌悬液混合后,得到混合溶液;所述载体溶液为海藻酸钠溶液、淀粉溶液、低聚果糖溶液、乳蔗糖溶液中的一种或多种;
(2)按照0.5~1.2L/min的通气量向混合溶液中持续通入空气后,将混合溶液滴入钙离子溶液中进行固化后制备得到益生菌胶珠。
2.如权利要求1所述的益生菌胶珠,其特征在于,步骤(1)中,所述载体溶液的浓度为10~50g/L;所述益生菌菌悬液与载体溶液添加的体积比为1:(8~10)。
3.如权利要求2所述的益生菌胶珠,其特征在于,步骤(2)中的钙离子溶液的质量分数为0.2~1.0%,进行固化的时间为30~50min。
4.如权利要求3所述的益生菌胶珠,其特征在于,所述益生菌胶珠外壁还涂覆有壳聚糖。
5.如权利要求4所述的益生菌胶珠,其特征在于,所述载体溶液为海藻酸钠溶液,所述海藻酸钠溶液的浓度为10~50g/L;
或所述载体溶液为低聚果糖-海藻酸钠溶液,所述低聚果糖-海藻酸钠溶液是将低聚果糖溶液和海藻酸钠溶液按照体积比为(1~1.5):1的比例混合后得到的,其中,所述低聚果糖溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L;
或所述载体溶液为淀粉-海藻酸钠溶液,所述淀粉-海藻酸钠溶液是将淀粉溶液和海藻酸钠溶液按照体积比为(1~1.5):1混合后得到的,其中,所述淀粉溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L;
或所述载体溶液为乳蔗糖-海藻酸钠溶液,所述乳蔗糖-海藻酸钠溶液是将乳蔗糖溶液和海藻酸钠溶液按照体积比为(1~1.5):1混合后得到的,其中,所述乳蔗糖溶液的浓度为10~50g/L,所述海藻酸钠溶液的浓度为10~50g/L。
6.如权利要求1~5任一所述的益生菌胶珠,其特征在于,所述益生菌包括但不限于,嗜酸乳杆菌(Lactobacillus acidophilus)、植物乳杆菌(Lactobacillus plantarum)、干酪乳杆菌(Lactobacillus casei)、保加利亚乳杆菌(Lactobacillus bulgaricus)、鼠李糖乳杆菌(Lactobacillus rhamnosus)和嗜热链球菌(Streptococcus thermophilus)中的一种或几种。
7.含有权利要求1~6任一所述的益生菌胶珠的产品。
8.如权利要求7所述的产品,其特征在于,所述产品为食品、药品或保健品。
9.权利要求1~5任一所述的益生菌胶珠,或权利要求7或8所述的产品在制备提高益生菌在胃肠道中活性的产品中的应用。
10.一种制备高活性益生菌胶珠的方法,其特征在于,所述包括以下步骤:
(1)将载体溶液与益生菌菌悬液混合后,得到混合溶液;所述载体溶液为海藻酸钠溶液、淀粉溶液、低聚果糖溶液、乳蔗糖溶液中的一种或多种;
(2)按照0.5~1.2L/min的通气量向混合溶液中持续通入空气后,将混合溶液滴入钙离子溶液中进行固化后制备得到益生菌胶珠。
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