CN112056399A - 一种增强免疫力的益生菌组合物及其应用 - Google Patents
一种增强免疫力的益生菌组合物及其应用 Download PDFInfo
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Abstract
本发明公开了一种增强免疫力的益生菌组合物及其应用。所述益生菌组合物包含益生元、K‑MYL益生菌组合和功能性物质,所述K‑MYL益生菌组合含有干酪乳杆菌,保加利亚乳杆菌,布拉迪酵母菌,罗伊氏乳杆菌,动物双歧杆菌中的至少一种,或者还包括鼠李糖乳杆菌、嗜酸乳杆菌、长双歧杆菌中的至少一种。本发明的益生菌组合物可以显著提升肠道有益菌群的数量,改善改善肠道免疫功能,提升肠道免疫因子的表达,抵抗病毒的感染以及缩短感染病毒后的恢复时间。而且本专利发明的益生菌株组合之间,以及益生元,益生菌菌株与功能性物质之间具有显著的协同增强作用。
Description
技术领域
本发明涉及肠道菌群调节领域,具体涉及通过调节肠道菌群增强免疫力,抵 抗病毒感染的口服益生菌组合物及其应用。
背景技术
人体肠道为微生物提供了良好的栖息环境,成人肠道内的微生物数量高达 1014,接近人体体细胞数量的10倍;质量达到1.2kg,接近人体肝脏的质量;其 包含的基因数目约是人体自身的100倍,具有人体自身不具备的代谢功能。作为 人体最庞大、最复杂的微生态系统,肠道微生物本身及其代谢产物不仅能调节人 体健康,更在膳食和宿主之间起到了重要的桥梁作用,是人体健康系统中非常重 要的组成部分。
肠道不仅是人体消化吸收的重要场所,同时也是最大的免疫器官,在维持正 常免疫防御功能中发挥着极其重要的作用。人体的健康与肠道内环境微生物有密 不可分的联系,正常身体状态下,肠道菌群通过与肠道的相互作用调控宿主的免 疫系统,维持动态平衡。一旦这种平衡被打破,肠道生态环境失衡,有害菌数量 增多,就会导致疾病。现代生活的疾病治疗中大量使用抗生素药物,以及食用含 抗生素或者农药等有毒物质残留的食物,严重危害着人类的肠道菌群生态健康, 有害菌比例上升产生有害物质,如产生诱发肝病的氨,引起肠道炎症的内毒素、 引发痛风,心血管疾病的尿酸,以及慢性肾病的对甲酚硫酸盐,从而导致慢性病 人越来越多,处于亚健康状态的人越来越多,非常容易被病毒或细菌感染生病, 甚至失去生命。在我国,每年因免疫力差,感染各种病毒(如流感病毒,轮状病毒,肺炎病毒,等)的人已达数亿人。
益生菌在调节肠道菌群生态的作用越来越被重视,大量的科学研究表明,益 生菌与肠道细胞具有紧密的联系和调控作用,诱导免疫因子的表达。而且最近的 许多研究结果表明,许多传统中草药的药效,亦是通过肠道菌群代谢后成为活性 物质发挥作用的。因为不同病人的肠道菌群不同,从而导致相同的中药方,在不 同的病人身上的效果会存在差异。此外,许多有增强免疫力作用的天然产物,如 香菇多糖,人参多糖,燕窝酸,壳寡糖,人参皂苷等,也与肠道益生菌以及肠道 免疫系统关系密切。调整菌群,重建肠道微生态,是一种安全的,无毒副作用的 提升亚健康人群的健康状况的方式。
然而,当前市场上的益生菌产品,多仅为补充益生菌为目标的,没有深入研 究益生菌株的功能与人体亚健康状态的关系以及益生菌菌株间的协同作用,又或 者仅把益生菌当做助消化或者增加风味的作用,如常温灭活酸奶,乳酸菌风味饮 料等,对人体健康的效果不显著,也远远低估了益生菌的作用。开发具有增强免 疫力的健康功能的益生菌产品,帮助亚健康人群恢复健康状态,可减少患病风险, 提升人民的生活质量,具有显著的社会意义,也具有巨大市场应用价值。
发明内容
针对上述现有技术的不足,本发明的目的在于提供一种通过调节肠道菌群增 强免疫力,抵抗病毒感染的益生菌组合物。本发明的益生菌组合物可以显著提升 肠道有益菌群的数量,改善肠道免疫功能,提升肠道免疫因子的表达,抵抗病毒 的感染以及缩短感染病毒后的恢复时间。而且本发明的益生菌株组合之间,以及 益生元,益生菌菌株与功能性物质之间具有显著的协同增强作用。
本发明的另一目的在于提供增强免疫力,抵抗病毒感染的益生菌组合物在制 备增强抗病毒感染能力的产品中的应用。
为实现上述目的,本发明的技术方案为:
本发明的第一方面,提供一种口服的增强免疫力的益生菌组合物,含有益生 元、K-MYL益生菌组合和功能性物质,所述K-MYL益生菌组合含有干酪乳杆 菌、保加利亚乳杆菌、布拉迪酵母菌、罗伊氏乳杆菌、动物双歧杆菌中的至少一 种。
优选的,所述K-MYL益生菌组合还包括鼠李糖乳杆菌、嗜酸乳杆菌、长双 歧杆菌中的至少一种。
优选的,所述益生菌组合物中的总活菌数≥1.0*108CFU/g,其中,干酪乳杆 菌、保加利亚乳杆菌、布拉迪酵母菌、罗伊氏乳杆菌、动物双歧杆菌中的单一益 生菌的活菌数≥2.0*107CFU/g,鼠李糖乳杆菌、嗜酸乳杆菌、长双歧杆菌中的单 一益生菌的活菌数≥0.5*107CFU/g。
优选的,所述益生元为低聚果糖、低聚半乳糖、低聚木糖、棉子糖、水苏糖 中的至少一种。
优选的,所述功能性物质为燕窝酸、壳寡糖、β-葡聚糖、针叶樱桃粉、猕 猴桃粉、维生素C中的至少一种。
进一步优选的,所述益生菌组合物含有低聚半乳糖、低聚木糖、干酪乳杆菌、 罗伊氏乳杆菌、保加利亚乳杆菌、布拉迪酵母菌、鼠李糖乳杆菌、长双歧杆菌、 嗜酸乳杆菌、动物双歧杆菌、β-葡聚糖、维生素C、猕猴桃水果粉;所述干酪乳 杆菌为保藏编号CGMCCNo.19708的干酪乳杆菌(Lactobacillus casei),罗伊氏乳 杆菌为保藏编号CGMCCNo.18700的罗伊氏乳杆菌(Lactobacillus reuteri),保加 利亚乳杆菌为保藏编号CGMCCNo.19709的保加利亚乳杆菌(Lactobacillus bulgaricus),鼠李糖乳杆菌为保藏编号CGMCC No.19711的鼠李糖乳杆菌 (Lactobacillus rhamnosus),动物双歧杆菌为保藏编号CGMCC No.19712的动物 双歧杆菌(Bifidobacterium animalis)。
第二方面,提供上述益生菌组合物在制备增强免疫力的产品中的应用。
优选的,上述应用中,所述增强免疫力的产品为食品、保健品、特医食品、 药品中的任一种。
优选的,上述应用中,所述增强免疫力的食品为固体饮料、酸奶、益生菌饮 料、酸奶块。
第三方面,提供上述益生菌组合物在制备抗病毒的产品中的应用。
优选的,上述应用中,所述抗病毒的产品为食品、保健品、药品中的任一种。
优选的,上述应用中,所述抗病毒的食品为固体饮料、酸奶、益生菌饮料、 酸奶块。
优选的,上述应用中,所述病毒为流感病毒。
在整个公开内容中,术语“K-MYL益生菌组合”为有助于增强免疫力的益 生菌组合。
本发明的有益效果:
本发明经过多轮的益生菌株功能的筛选和菌株组合优化,辅配协同增强益生 菌效果的功能性物质,并在动物体内和受试人群验证其健康功能效果。本发明的 益生菌组合物较常规的市售益生菌粉,其生长效率最高增加600%以上,可以显 著提升肠道有益菌群的数量。经动物试验验证,本发明的益生菌组合物可显著提 高小鼠的NK细胞活性和淋巴细胞转化能力,增强免疫力,且多种益生菌组合的 效果优于单一益生菌效果,证明本发明的益生菌组合之间具有协同增强的效果。 受试人群服用后,免疫球蛋白含量提高,减少流感病毒感染的风险,并缩短感染 流感后的恢复时间。本专利发明的益生菌株组合物,可以缓解当下社会中快节奏, 高压力生活节奏下带来的免疫力下降的亚健康状态,增强抵抗病毒感染的能力, 具有巨大的社会意义和经济价值。
生物保藏说明
干酪乳杆菌(Lactobacillus casei),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址北京市朝阳区北辰西路1号院3号,保藏机构简称: CGMCC,保藏日期为2020年04月23日,生物保藏编号为CGMCC No.19708, 菌株编号:KLca-10。
罗伊氏乳杆菌(Lactobacillus reuteri),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址北京市朝阳区北辰西路1号院3号,保藏机构简称: CGMCC,保藏日期为2019年10月17日,生物保藏编号为CGMCC No.18700, 菌株编号:KLR-3。
保加利亚乳杆菌(Lactobacillus bulgaricus),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址北京市朝阳区北辰西路1号院3号,保藏机 构简称:CGMCC,保藏日期为2020年04月23日,生物保藏编号为CGMCC No.19709,菌株编号:KLdB-1。
鼠李糖乳杆菌(Lactobacillus rhamnosus),保藏于中国微生物菌种保藏管理 委员会普通微生物中心,保藏地址北京市朝阳区北辰西路1号院3号,保藏机构 简称:CGMCC,保藏日期为2020年04月23日,生物保藏编号为CGMCC No.19711,菌株编号:KLrh-10。
动物双歧杆菌(Bifidobacterium animalis),保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址北京市朝阳区北辰西路1号院3号,保藏机构 简称:CGMCC,保藏日期为2020年04月23日,生物保藏编号为CGMCC No.19712,菌株编号:KBla-22。
具体实施方式
通过以下详细说明可以进一步理解本发明的特点和优点。所提供的实施例仅 是对本发明方法的说明,而不以任何方式限制本发明揭示的其余内容。
以下实施例中所用到的原料和益生菌粉,除特别注明外,均为市售产品或湖 北康肽药业有限公司生产的益生菌粉。功效性性物质添加量参照了相应的国家标 准:GB 2760-2014《食品添加剂使用标准》;GB 14880-2012《食品营养强化剂使 用标准》;相应的行业标准和国家卫生部等主管部门颁布的最新的新食品原料使 用规范或公告等,配料添加量均符合相应的法规规定。
以下实施例中干酪乳杆菌,保加利亚乳杆菌,布拉迪酵母菌,罗伊氏乳杆菌, 动物双歧杆菌中的单一益生菌的活菌数≥2.0*107CFU/g,鼠李糖乳杆菌、嗜酸乳 杆菌、长双歧杆菌中的单一益生菌的活菌数≥0.5*107CFU/g。
实施例1:筛选对K-MYL复合益生菌的促进作用益生元
本实施例针对K-MYL益生菌组合进行了促增长益生元的筛选,从而获得促 进益生菌生长的益生元组合。以MRS培养基(牛肉膏10.0g,酵母膏5.0g,蛋 白胨10.0g,葡萄糖20.0g,K2HPO4 2.0g,醋酸钠5.0g,MgSO4·7H2O 0.2g, MnSO4·4H2O 0.05g,吐温80 1.0g,柠檬酸三铵2.0g,琼脂15.0g,加蒸馏水至 1000ml)为基础培养基,分别添加候选的益生元或益生元组合至终浓度为10%。 按相同的接种量接入K-MYL复合益生菌冻干粉(含布拉迪酵母菌粉35%,干酪 乳杆菌粉15%,保加利亚乳杆菌10%,动物双歧杆菌粉10%,鼠李糖乳杆菌粉 15%,长双歧杆菌粉10%,嗜酸乳杆菌粉5%),厌氧培养箱静置培养6h,测定 菌体生长的OD600值,筛选促进益生菌生长的益生元组合物。筛选结果(表1) 显示,菊粉(低聚果糖)、低聚半乳糖、低聚木糖、棉子糖、水苏糖等益生元的 促进益生菌生长的效果较好,促进效果较基础培养基提高105%-629%,可提高 在肠道内的益生菌数量。
表1.促进K-MYL复合益生菌生长的益生元筛选
实施例2:增强免疫力的固体饮料
本实施例提供一种能够维持肠道健康和增强免疫力的益生菌固体饮料,各辅 料的组成如表2所示,精确称量原料后,进行梯度混匀,最终分装为10g/袋。经 检测(参考GB4789.35-2016方法描述),除不含益生菌的产品10外,其余固体 饮料的益生菌活菌数为1.8-8.2×108CFU/g。
表2.增强免疫力的固体饮料
实施例3:增强免疫力的酸奶
本实施例提供一种能够维持肠道健康和增强免疫力的功能益生菌发酵酸奶, 是由下列重量的组分组成:鲜牛乳100g,白砂糖12g,酵母β-葡聚糖250mg, 维生素C 200mg,95℃灭菌5min后冷却至40℃,接种K-MYL复合益生菌发酵 剂0.18g(嗜热链球菌冻干粉20%,保加利亚乳杆菌冻干粉15%,干酪乳杆菌冻 干粉20%,鼠李糖乳杆菌冻干粉20%,动物双歧乳杆菌冻干粉15%,嗜酸乳杆 菌冻干粉10%),密封后置于酸奶机在40℃发酵7-12h,然后置于4℃冰箱中12h。 在洁净环境下,加入1%的布拉迪酵母菌粉,搅拌均匀后分装,4℃条件下储存。 经检测(乳酸菌参考GB4789.35-2016方法描述,酵母菌参考GB/T 22547方法描述),该酸奶的乳酸菌活菌数为2.2×108CFU/g,布拉迪酵母活菌数为3.4×107 CFU/g。
实施例4:增强免疫力的益生菌饮料
本实施例提供一种能够维持肠道健康和缓解抑郁/焦虑情绪的功能性益生菌 饮料,是由下列重量的组分组成:每100mL的益生菌饮料中含有脱脂乳粉10g, 食用白砂糖8g,5g针叶樱桃粉,100mg维生素C,和K-MYL益生菌0.82g(20% 干酪乳杆菌冻干粉、20%布拉迪酵母菌冻干粉,10%保加利亚乳杆菌冻干粉,10% 长双歧杆菌冻干粉、10%鼠李糖乳杆菌冻干粉、10%的罗伊氏乳杆菌冻干粉,10% 嗜酸乳杆菌冻干粉、10%动物双歧杆菌冻干粉)。经检测(乳酸菌参考 GB4789.35-2016方法描述,酵母菌参考GB/T 22547方法描述),益生菌饮料的 活菌量为2.5×108CFU/g,布拉迪酵母活菌数为3.5×107CFU/g。
实施例5:增强免疫力的酸奶块
本实施例提供一种能够维持肠道健康和提高免疫力的功能性益生菌饮料,首 先是酸奶发酵,将鲜牛乳1000g,白砂糖90g混合均匀,加热至80-90℃维持10 分钟,降温至40℃以下,加入K-MYL复合益生菌发酵剂1g(嗜热链球菌冻干 粉20%,保加利亚乳杆菌冻干粉15%,干酪乳杆菌冻干粉20%,罗伊氏乳杆菌 冻干粉10%,鼠李糖乳杆菌冻干粉20%,长双歧乳杆菌冻干粉5%,嗜酸乳杆菌 冻干粉10%)并搅拌均匀,密封后置于酸奶机在40℃发酵7-12h使酸奶凝固, 然后置于4℃冰箱中12h。将酸奶与辅料按以下重量分混合:酸奶30%、白砂糖 7%、奶粉7%、乳清粉5%、柠檬酸6%、柠檬酸钠3%,布拉迪酵母粉0.5%,淀 粉1%、果胶1%、β-葡聚糖5%,水24.5%、香蕉水果粒10%。所有物料混合均 匀后,倒入乳化罐中乳化30分钟,装入酸奶块制备不锈钢格板,放入冷冻干燥 机冻干,冻干程序如表3所示。经检测(乳酸菌参考GB4789.35-2016方法描述, 酵母菌参考GB/T 22547方法描述),益生菌饮料的活菌量为15.5×108CFU/g, 布拉迪酵母活菌数为6.4×108CFU/g。
表3.益生菌酸奶块冻干程序
实施例6:提高免疫力试验
100只SPF昆明种小白鼠,雌雄各半,6-8周龄,体重18-22g。受试样品为实 施例2-5中的样品,人体推荐剂量为10g/天(以体重60kg计),在该试验中受试样品 剂量为1.7g/kg·bw/d(相当于受试样品人体推荐摄入量的10倍),以奶粉代替受试 物为空白对照。受试样品用无菌水配制,配制浓度分别为500g/L,经口每日分三 次给予小鼠相应剂量的受试物,小鼠总灌胃量为3.4mL/kg·bw,连续灌胃30天。 采用胸腺/体重比值、脾脏/体重比值,ConA诱导的小鼠脾淋巴细胞转化试验(MTT 法)、NK细胞活性测定(LDH法)的方式对本发明产品进行增强免疫力功能验证。 试验结果(表4)显示,产品对小鼠胸腺和脾脏的重量没有显著影响(p>0.05)。 免疫力测试结果(表5)显示,实施例2-5的受试样品组与空白对照组相比,均可 显著提高小鼠溶血空斑数,表明本发明的益生菌组合物具有增强小鼠体液免疫功能,同时小鼠NK细胞活性提高表明,益生菌颗粒具有显著增强小鼠NK细胞活性 作用。实施例2中的产品1、2、6、8、10的受试结果显示,益生菌株组合的效果 优于单一益生菌效果,益生元,益生菌和功能物质的组合(产品1)优于益生元 与益生菌组合(产品9)或益生元与功能物质组合(产品10)的受试组,说明益 生元,益生菌和功效物质具有协同增强作用。
表4.不同测试组的小鼠胸腺/体重和脾脏/体重比值
注:与对照组比较,*:p<0.05;#:p<0.01
表5.ConA诱导的小鼠脾淋巴细胞转化试验
注:与对照组比较,*:p<0.05;#:p<0.01
实施例7:增强免疫力,抗流感病毒的人体效果试验
选取100名中老年人(年龄50-70岁),分为5组,分别服用按实施例2-5配方 生产的固体饮料(产品1),发酵酸奶,益生菌饮料,酸奶块,每日1次,固体饮 料和酸奶块每次10g,发酵酸奶和益生菌饮料每次100ml;对照组不做干预,持续 6个月(9月至次年2月份)。干预前后,分别测定血液生化,比较其体液抗体水平。 同时统计不同测试组这6个月内的流感发病率,以及感冒后的康复时间。
表6.受试者干预前后的免疫球蛋白变化
表7.不同产品的受试者在干预后的感冒发生率及恢复时间
表6与表7的结果显示,与未做干预的对照组相比,服用了本发明实例的含 K-MYL益生菌的产品,显著提升了血液免疫球蛋白的数量,且明显降低了感冒 的发病率,其发病率能够降至10%以下,说明本发明提供的增强免疫力的含益 生菌组合物配方制备的产品,能够增强免疫力,有效降低流感病毒引起感冒几率, 同时缩短康复时间。
Claims (10)
1.一种口服的增强免疫力的益生菌组合物,其特征在于,包含益生元、K-MYL益生菌组合和功能性物质,所述K-MYL益生菌组合含有干酪乳杆菌、保加利亚乳杆菌、布拉迪酵母菌、罗伊氏乳杆菌、动物双歧杆菌中的至少一种。
2.根据权利要求1所述的益生菌组合物,其特征在于,所述益生元为低聚果糖、低聚半乳糖、低聚木糖、棉子糖、水苏糖中的至少一种。
3.根据权利要求1所述的益生菌组合物,其特征在于,所述功能性物质为燕窝酸、壳寡糖、β-葡聚糖、针叶樱桃粉、猕猴桃粉、维生素C中的至少一种。
4.根据权利要求1所述的益生菌组合物,其特征在于,所述K-MYL益生菌组合还包括鼠李糖乳杆菌、嗜酸乳杆菌、长双歧杆菌中的至少一种。
5.根据权利要求4所述的益生菌组合物,其特征在于,所述益生菌组合物中的总活菌数≥1.0*108CFU/g,其中,干酪乳杆菌、保加利亚乳杆菌、布拉迪酵母菌、罗伊氏乳杆菌、动物双歧杆菌中的单一益生菌的活菌数≥2.0*107CFU/g,鼠李糖乳杆菌、嗜酸乳杆菌、长双歧杆菌中的单一益生菌的活菌数≥0.5*107CFU/g。
6.根据权利要求5所述的益生菌组合物,其特征在于,所述益生菌组合物含有低聚半乳糖、低聚木糖、干酪乳杆菌、罗伊氏乳杆菌、保加利亚乳杆菌、布拉迪酵母菌、鼠李糖乳杆菌、长双歧杆菌、嗜酸乳杆菌、动物双歧杆菌、β-葡聚糖、维生素C、猕猴桃水果粉;所述干酪乳杆菌为保藏编号CGMCC No.19708的干酪乳杆菌(Lactobacillus casei),罗伊氏乳杆菌为保藏编号CGMCC No. 18700的罗伊氏乳杆菌(Lactobacillus reuteri),保加利亚乳杆菌为保藏编号CGMCC No.19709的保加利亚乳杆菌(Lactobacillus bulgaricus),鼠李糖乳杆菌为保藏编号CGMCC No.19711的鼠李糖乳杆菌(Lactobacillus rhamnosus),动物双歧杆菌为保藏编号CGMCC No.19712的动物双歧杆菌(Bifidobacterium animalis)。
7.权利要求1-6任一项所述的的益生菌组合物在制备在制备增强免疫力和/或抗病毒的产品中的应用。
8.根据权利要求7所述的应用,其特征在于,所述的产品为食品、保健品、特医食品、药品中的任一种。
9.根据权利要求8所述的应用,其特征在于,所述的食品为固体饮料、酸奶、益生菌饮料、酸奶块。
10.根据权利要求7所述的应用,其特征在于,所述病毒为流感病毒。
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