CN115299494A - 乳双歧杆菌Probio-M8在改善儿童肠道菌群中的应用 - Google Patents
乳双歧杆菌Probio-M8在改善儿童肠道菌群中的应用 Download PDFInfo
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- CN115299494A CN115299494A CN202110491585.1A CN202110491585A CN115299494A CN 115299494 A CN115299494 A CN 115299494A CN 202110491585 A CN202110491585 A CN 202110491585A CN 115299494 A CN115299494 A CN 115299494A
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- Dairy Products (AREA)
Abstract
本发明涉及微生物技术领域,具体涉及乳双歧杆菌Probio‑M8在改善儿童肠道菌群中的应用。本发明提供乳双歧杆菌Probio‑M8在制备用于靶向调控儿童特征产短链脂肪酸肠道菌的产品中的应用,所述乳双歧杆菌Probio‑M8的保藏编号为CGMCC No.18610。本发明通过对儿童和成人肠道菌群组成进行分析,确定了儿童特征产SCFA肠道菌,针对儿童特征产SCFA肠道菌,发现乳双歧杆菌Probio‑M8具有靶向、精准调控儿童特征产SCFA肠道菌的功能,可用于改善儿童肠道菌群,增强肠道屏障功能,促进儿童肠道健康。
Description
技术领域
本发明涉及微生物技术领域,具体涉及乳双歧杆菌Probio-M8在改善儿童肠道菌群中的应用、乳双歧杆菌组合物以及产品。
背景技术
乳双歧杆菌(Bifidobacterium lactis)属于双歧杆菌属细菌,是定植于人体肠道内的有益菌,具有调节肠道菌群、促进营养物质消化吸收等多种功能。
肠道菌群与人体健康息息相关。肠道菌群可发酵不可消化的碳水化合物(膳食纤维)产生短链脂肪酸(short-chain fatty acids,SCFAs)。SCFAs是含1-6个碳原子的有机羧酸,包含乙酸、丙酸、丁酸、异丁酸等。90%的SCFAs是由人体内肠道菌群发酵不可消化的碳水化合物产生的,每日产生的SCFAs可达到50-100mmol。肠道SCFAs中含量较高的为丁酸、乙酸、丙酸,三者比例约为60:20:20。SCFAs通过神经-内分泌-免疫调节,直接或间接介导微生物肠脑轴,对宿主的生理功能产生重要影响。SCFAs还参与肠道能量供应,可影响肠腔pH、肠黏膜屏障的通透性,还可发挥调节免疫、抗炎、提高胰岛素敏感性、改善肥胖等生理功能。其中,乙酸是胆固醇合成的底物,丙酸影响脂肪生成和糖异生,丁酸是结肠细胞的主要能量来源,可提高胰岛素敏感性,并发挥抗炎、抗肥胖作用。常见的产SCFASs菌群包括:拟杆菌属、梭菌属、双歧杆菌属、真杆菌属、链球菌属、消化链球菌属等。其中,拟杆菌和双歧杆菌主要产生乙酸盐和丙酸盐,厚壁菌产生丁酸盐,不同SCFAs的产生量受厚壁菌和拟杆菌比例的影响。
SCFA合成的最重要手段是肠道细菌的内源性发酵,因此,不同人群特征产SCFA菌群的多样性及体内不可消化的碳水化合物种类决定了其内源性SCFA种类和量效。
目前采用的调节体内SCFA的方法主要包括:直接口服乙酸盐、丁酸盐等;补充低聚果糖、低聚半乳糖等益生元;补充普通双歧杆菌、乳酸杆菌等益生菌;食用不可消化的膳食纤维或食材。
现有的调节体内SCFA的手段多是对不同人群施用同样的方法,未针对特定人群的特征产SCFA肠道菌群进行精准施策。与成人肠道菌群不同,儿童肠道菌群仍处于过渡时期,容易受到营养水平、疾病、环境因素的影响。而且,儿童和成人的产SCFA肠道菌群的组成具有明显差异,但是,目前尚未有针对儿童特征产SCFA肠道菌群的特点进行精准调节的益生菌及其产品。
发明内容
本发明的目的是提供乳双歧杆菌Probio-M8在改善儿童肠道菌群中的应用。本发明的另一目的是提供含有乳双歧杆菌Probio-M8的产品、乳双歧杆菌组合物和食品,该组合物、产品和食品可针对儿童特征产SCFA肠道菌群的特点靶向调控儿童特征产SCFA肠道菌群。
本发明以开发针对儿童特征产SCFA肠道菌进行靶向、精准调节的益生菌为目的,首先对儿童肠道特征菌群进行了分析,发现了中国儿童特征产SCFA肠道菌群,其与成人产SCFA肠道菌群存在明显差异。进一步针对儿童特征产SCFA肠道菌,本发明对大量的益生菌进行筛选,获得了能够高效特异性富集儿童产SCFA肠道菌群的益生菌——乳双歧杆菌Probio-M8。本发明发现,与其他常用益生菌相比,乳双歧杆菌Probio-M8能够更有效地调控儿童肠道菌群,特异性富集儿童特征产SCFA肠道菌群。
乳双歧杆菌Probio-M8菌株已在发明专利CN 110892935 A中公开,该菌株分离自中国健康妇女母乳,是一株具有潜在益生特性的双歧杆菌。该菌株已于2019年9月20日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编100101),分类命名为乳双歧杆菌Bifidobacterium lactis,保藏编号为CGMCC No.18610。
具体地,本发明提供以下技术方案:
本发明首先提供乳双歧杆菌Probio-M8在制备用于靶向调控儿童特征产短链脂肪酸肠道菌的产品中的应用,所述乳双歧杆菌Probio-M8的保藏编号为CGMCC No.18610。
本发明所述的靶向调控具体为靶向富集。
优选地,所述儿童特征产短链脂肪酸肠道菌为产乙酸肠道菌、产丁酸肠道菌、产丙酸肠道菌中的一种或多种。
乳双歧杆菌Probio-M8可特异性富集儿童特征产乙酸肠道菌、产丁酸肠道菌和产丙酸肠道菌,尤其对于产乙酸肠道菌的富集效果更优。
优选地,所述儿童特征产短链脂肪酸肠道菌包括选自以下属中的一种或多种:双歧杆菌(Bifidobacterium)、链球菌(Streptococcus)、瘤胃球菌(Ruminococcus)和真杆菌(Eubacterium)。
本发明发现,与成年人的产短链脂肪酸肠道菌明显不同,双歧杆菌(Bifidobacterium)、链球菌(Streptococcus)、瘤胃球菌(Ruminococcus)和真杆菌(Eubacterium)是中国儿童特征产短链脂肪酸肠道菌,而乳双歧杆菌Probio-M8能够高效、特异地富集双歧杆菌(Bifidobacterium)、链球菌(Streptococcus)、瘤胃球菌(Ruminococcus)和真杆菌(Eubacterium)中的产短链脂肪酸肠道菌。
本发明发现,在上述产短链脂肪酸肠道菌中,乳双歧杆菌Probio-M8对于产乙酸肠道菌Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacterium dentium、Bifidobacterium hydrogenotrophica、Streptococcus agalactiae、Streptococcusdysgalactiae、Streptococcus thermophilus、Streptococcus urinalis、Streptococcusvestibularis、Bifidobncterium pseudongum、Bifidobncterium longum,产丙酸肠道菌Bifidobacterium adolescentis、Eubacterium hallii Ruminococcus torques、Ruminococcus bromii以及产丁酸肠道菌Eubacterium hallii、Subdoligranulumvariabile、Ruminococcus torques、butyrate-producing bacterium SSC/2、Eubacteriumdolichum的富集作用十分显著。
进一步优选地,所述儿童特征产短链脂肪酸肠道菌为选自Bifidobacteriumbifidum、Bifidobacterium breve、Bifidobacterium dentium、Bifidobacteriumhydrogenotrophica、Streptococcus agalactiae、Streptococcus dysgalactiae、Streptococcus thermophilus、Streptococcus urinalis、Streptococcus vestibularis、Bifidobncterium pseudongum、Bifidobncterium longum、Bifidobacteriumadolescentis、Ruminococcus torques、Ruminococcus bromii、Eubacterium hallii、Subdoligranulum variabile、butyrate-producing bacterium SSC/2、Eubacteriumdolichum中的一种或多种。
在上述产短链脂肪酸肠道菌中,乳双歧杆菌Probio-M8尤其对Bifidobacteriumbifidum、Streptococcus thermophilus、Streptococcus urinalis、Bifidobacteriumadolescentis的特异性富集效果更优。
本发明通过实验验证发现,乳双歧杆菌Probio-M8可单独发挥靶向调控儿童特征产短链脂肪酸肠道菌的功能。
基于此,本发明提供一种含有乳双歧杆菌Probio-M8的产品,所述乳双歧杆菌Probio-M8的保藏编号为CGMCC No.18610。
具体地,所述含有乳双歧杆菌Probio-M8的产品包括冷冻饮品、乳制品、调味品、焙烤品、肉制品、糖果或涂抹食品。
所述冷冻饮品包括冰淇淋、雪糕等。
所述乳制品包括牛奶、乳粉、酸牛奶、发酵乳、乳饮料、奶酪、奶片、黄油、稀奶油、生奶油等。
所述焙烤品包括面包、蛋糕、甜面团产品、叠层面团产品、液态奶蛋糊、松饼、油炸圈饼、饼干、薄脆饼干、曲奇饼等。
所述糖果包括巧克力、饴糖、胶皮糖、布丁等。
所述肉制品包括经加工的肉制品。
所述涂抹食品为用于食品的涂抹物。
具体地,以上所述的产品为发酵乳制品,所述发酵乳制品的原料包含如下重量份的组分:乳原料80-95份,乳双歧杆菌Probio-M80.001-0.5份、108-1011cfu/100g原料。
具体地,以上所述的产品为调制乳粉,其由包括如下重量份的原料制备得到:生牛乳320-355份,脱脂乳粉25.8-28.6份,浓缩乳清蛋白粉0.48-0.55份,加糖炼乳23.5-26份,磷脂0.56-0.63份,大豆油2.9-3.3份,低聚果糖3.1-3.5份,低聚半乳糖0.73-0.82份,维生素预混料0.2-0.27份,(3R,3’R)-二羟基-β-胡萝卜素(玉米黄质)0.0089-0.01份,七水硫酸亚铁0.039-0.044份,七水硫酸锌0.029-0.033份,四水硫酸镁0.23-0.26份,碳酸钙0.94-1.04份,5%硫酸铜0.029-0.033份,富硒酵母0.0075-0.009份,氯化胆碱0.094-1.04份,5%叶黄素0.0043-0.0048份,酵母β-葡聚糖0.01-0.36份,二十二碳六烯酸0-0.65份,花生四烯酸0-1.3份,乳铁蛋白0-0.11份,磷脂酰丝氨酸0-1.0份,N-乙酰神经氨酸0-0.5份,异构化乳糖0-2.21份,柠檬酸0.01-0.43份,氢氧化钾0.001-0.104份,乳双歧杆菌Probio-M8 0.001-0.6份、5×108-5×1010cfu/100g原料;
所述维生素预混料包括:醋酸维生素A、维生素D3、DL-α-醋酸生育酚、植物甲萘醌、盐酸硫胺素、核黄素、盐酸吡哆醇、L-抗坏血酸、D-泛酸钙、D-生物素、叶酸、烟酰胺、氰钴胺和牛磺酸。
本发明发现上述配方的发酵乳制品和调制乳粉具有更优的靶向调控儿童特征产短链脂肪酸肠道菌的功能。
以上所述的发酵乳制品和调制乳粉可采用本领域常规方法制备得到。
具体地,所述发酵乳制品的制备方法可包括如下步骤:将除菌种外的原料均质,均质后进行杀菌,冷却后进行接种,加入菌种,于40-42℃发酵,至pH为4.2-4.7结束发酵,进行破乳处理并包装成相应规格的产品。
具体地,所述调制乳粉的制备方法可包括如下步骤:将乳原料、碳水化合物原料和预混的油脂原料混合后均质,经冷却后加入维生素和矿物质,经杀菌和浓缩后进行喷雾干燥,经冷却过筛进行充氮包装,待检测合格后与小料混合并包装成相应规格的产品。
另一方面,本发明还发现,将乳双歧杆菌Probio-M8与益生元配合使用,能够更好地促进乳双歧杆菌Probio-M8发挥靶向富集儿童特征产短链脂肪酸肠道菌的作用。
基于此,本发明提供一种乳双歧杆菌组合物,其包括乳双歧杆菌Probio-M8和益生元,所述乳双歧杆菌Probio-M8的保藏编号为CGMCC No.18610;所述益生元为选自菊粉、低聚半乳糖、低聚果糖、低聚异麦芽糖、低聚木糖、低聚甘露糖、半乳甘露聚糖、水苏糖、聚葡萄糖、抗性糊精中的一种或多种。
本发明进一步发现,与其他益生元相比,将菊粉与乳双歧杆菌Probio-M8配合使用,能够更好地促进乳双歧杆菌Probio-M8发挥靶向富集儿童特征产短链脂肪酸肠道菌的作用。
本发明尝试将不同链长的菊粉与乳双歧杆菌Probio-M8组合使用,并发现,乳双歧杆菌Probio-M8与不同链长的菊粉组合,对于儿童特征产短链脂肪酸肠道菌均具有一定的靶向调控作用。
本发明进一步发现,与其他聚合度的菊粉相比,聚合度为2-10的短链菊粉更容易被乳双歧杆菌Probio-M8利用,且能够更好地促进乳双歧杆菌Probio-M8发挥靶向调控儿童特征产短链脂肪酸肠道菌的作用,将聚合度为2-10的菊粉与乳双歧杆菌Probio-M8组合使用,针对儿童特征肠道菌群的靶向、精准调控作用效果明显更优。
优选地,所述益生元为菊粉,所述菊粉的聚合度为2-10。
本发明所述的聚合度为2-10的菊粉可为在该聚合度范围内的任意聚合度菊粉的混合物。
以上所述的乳双歧杆菌组合物中,乳双歧杆菌Probio-M8与菊粉的比例为108-1011cfu:(0.2-2)g,优选为108-1010cfu:(0.5-1)g。
以上乳双歧杆菌组合物可采用本领域常规方法制备得到。其中,乳双歧杆菌Probio-M8的培养可在MRS等常用双歧杆菌培养基中,于37℃厌氧培养。
本发明提供的上述含有乳双歧杆菌Probio-M8的组合物可直接添加于食品、保健品或药物中,或者,作为原料用于制备食品、保健品或药物。基于其特异性精准调控儿童产SCFA肠道菌群的优势,该组合物更适合应用于儿童食品、保健品或药品。
本发明还提供上述乳双歧杆菌组合物在食品、保健品或药物制备中的应用。
本发明所述的食品包括但不限于乳制品,冷冻饮品,可可制品、巧克力和巧克力制品,糖果,焙烤食品,蛋制品,饮料类食品,脂肪、油和乳化脂肪制品,肉制品等。
其中,乳制品包括牛奶、乳粉、酸奶、凝乳、发酵乳、乳饮料、奶酪、奶片、黄油、稀奶油、生奶油等。冷冻饮品包括冰淇淋、雪糕等。焙烤品包括面包、蛋糕、甜面团产品、叠层面团产品、液态奶蛋糊、松饼、油炸圈饼、饼干、薄脆饼干、曲奇饼等。糖果包括饴糖、胶皮糖、布丁、果冻等。
进一步地,本发明提供一种食品,所述食品包含所述乳双歧杆菌组合物,或者,所述食品的制备原料包含所述乳双歧杆菌组合物。
作为本发明的一种实施方式,所述食品为发酵乳制品,所述发酵乳制品的原料包含如下重量份的组分:乳原料80-95份,乳双歧杆菌Probio-M8 0.001-0.5份、108-1011cfu/100g原料,菊粉0.2-2份。
优选地,所述乳原料为选自生牛乳、复原乳中的一种或两种。
以上所述的发酵乳制品原料还包括糖类和蛋白粉。
优选地,本发明提供的发酵乳制品的原料包含如下重量份的组分:乳原料80-95份,糖类3-15份,蛋白粉0.1-5份,乳双歧杆菌Probio-M8 0.001-0.05份、108-1011cfu/100g原料,菊粉0.2-2份。
本发明发现上述配方的发酵乳制品具有更优的靶向调控儿童特征产短链脂肪酸肠道菌的功能。
作为本发明的优选方案,本发明提供的发酵乳制品的原料包含如下重量份的组分:生牛乳85-95份,白砂糖5-10份,乳双歧杆菌Probio-M8 0.001-0.05份、108-1011cfu/100g原料,菊粉0.5-1份。
本发明所述的发酵乳制品中,乳双歧杆菌Probio-M8的浓度为106-1010cfu/g。乳双歧杆菌Probio-M8在该含量范围内,发酵乳制品对于儿童肠道菌群和健康状况的改善作用更优。
本发明还提供所述发酵乳制品的制备方法,其包括对所述原料进行发酵的步骤,具体地,所述发酵为在41-43℃发酵,至pH为4.2-4.7结束发酵。
以上所述的发酵乳制品的制备方法具体包括如下步骤:将除菌种外的原料均质,均质后进行杀菌,冷却后进行接种,加入菌种,于40-42℃发酵,至pH为4.2-4.7结束发酵,进行破乳处理并包装成相应规格的产品。
本发明的有益效果在于:本发明通过对儿童和成人肠道菌群组成进行分析,发现了儿童特征产SCFA肠道菌。针对儿童特征产SCFA肠道菌,发现乳双歧杆菌Probio-M8具有靶向、精准调控儿童特征产SCFA肠道菌的功能,乳双歧杆菌Probio-M8能够特异性高效富集儿童特征产SCFA肠道菌,用于改善儿童肠道菌群,增强肠道屏障功能,促进儿童肠道健康。
在乳双歧杆菌Probio-M8的基础上,本发明还开发了能够靶向调控儿童特征产SCFA肠道菌的含乳双歧杆菌Probio-M8的发酵乳、调制乳粉、以及乳双歧杆菌Probio-M8的组合物和含有该组合物的发酵乳制品,这些产品均具有优异的靶向富集儿童特征产SCFA肠道菌的功能。
附图说明
图1为本发明实施例1中的中国儿童特征肠道菌群(种水平),其中,经过平方根转换的相对丰度差异倍数的log2值,正值表示在儿童中丰度高,负值表示在成年人中丰度高。
图2为本发明实施例1中的中国儿童特征肠道菌群(种水平),其中,经过平方根转换的相对丰度差异倍数的log2值,正值表示在儿童中丰度高,负值表示在成年人中丰度高。
图3为本发明实施例1中乳双歧杆菌Probio-M8干预后中国儿童特征产乙酸代谢肠道菌的丰度。
图4为本发明实施例1中乳双歧杆菌Probio-M8干预后中国儿童特征产丙酸代谢肠道菌的丰度。
图5为本发明实施例1中乳双歧杆菌Probio-M8干预后中国儿童特征产丁酸代谢肠道菌的丰度。
图6为本发明实施例2中乳双歧杆菌Probio-M8干预后中国儿童特征产乙酸代谢肠道菌的丰度。
图7为本发明实施例2中乳双歧杆菌Probio-M8干预后中国儿童特征产丙酸代谢肠道菌的丰度。
图8为本发明实施例2中乳双歧杆菌Probio-M8干预后中国儿童特征产丁酸代谢肠道菌的丰度。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。
实施例1乳双歧杆菌Probio-M8对儿童特征产SCFA肠道菌的富集作用检测(1)
本实施例首先制备了含乳双歧杆菌Probio-M8的益生菌酸奶,用于检测乳双歧杆菌Probio-M8对儿童特征产SCFA肠道菌的富集作用。
益生菌酸奶的原料由如下组分组成:菊粉(聚合度为2-10)0.7g,白砂糖7.5g,蛋白粉1.0g,乳双歧杆菌Probio-M8 0.0025g(含乳双歧杆菌Probio-M8 5×108cfu),生牛乳补足至100g。
该益生菌酸奶产品中乳双歧杆菌Probio-M8的含量为5×106cfu/g酸奶。
该益生菌酸奶的制备方法具体如下:将除乳双歧杆菌之外的所有原料混合后,于65℃预热,在40-160bar压力条件下均质后,于95℃杀菌5min,冷却至41±1℃,加入菌种,42℃发酵,至pH4.5-4.6时破乳,冷却至20℃。
对于儿童特征产SCFA肠道菌的富集作用检测的实验方法如下:
招募儿童志愿者共106名,招募标准为:年龄8-9岁的中国学龄儿童;一个月内未使用过益生菌、益生元制剂或抗生素,且入组时无呼吸道和胃肠道疾病症状。将志愿者分为2组,其中,对照组49人,每日正常膳食;干预组57人,除正常膳食外,每日饮用上述益生菌酸奶产品,每日饮用1次,每次服用量为200mL,干预周期为40天。
在干预前和干预结束后,分别采集志愿者的粪便样品,利用Illumina Miseq高通量测序平台进行粪便样品的宏基因组测序,通过属水平和种水平物种富集分析,获得肠道菌群数据。
1、儿童特征产SCFA肠道菌群分析
将对照组儿童的肠道菌群数据与随机选取的相同数量的健康成年人的肠道菌群数据进行比较,分析儿童人群的肠道特征菌群(Adjusted p value<0.01)。
属水平的分析结果如表1所示,结果显示,Bacteroides、Prevotella、Faecalibacteriuim均为中国成人和儿童肠道菌群丰度的前三位,而Ruminococcus、Blautia和Bifidobacterium是中国儿童特征的肠道菌群,Escherichia、Parabacteroides和Clostridium中国成人特征性肠道菌群。
表1儿童特征产SCFA肠道菌群属水平分析结果
种水平的分析结果如图1和图2所示,结果显示,Bifidobacterium longum、Bifidobacterium adolescentis、Bifidobacterium dentium、Bifidobacteriumcatenulatum、Bifidobacterium pseudocatenulatumcomplex、Lactobacillus reuteri等有益菌是中国儿童特征性的肠道菌群,而Fusobacterium varium、Klebsiellapneumoniae、Klebsiella variicola、Enterobacter aerogenes等是中国成人特征性肠道菌群。
儿童特征产SCFA肠道菌群分析结果如表2所示,结果显示,Bifidobacterium、Streptococcus、Ruminococcus和Eubacterium是中国儿童特征性产SCFA肠道菌群,而Bacteroides、Prevotella和Megasphaera是中国成人特征性产SCFA肠道菌群。
表2中国儿童和成人特征性产SCFA肠道菌群
2、酸奶对儿童产SCFA肠道菌群的选择性富集作用
利用Illumina Miseq高通量测序平台进行粪便样品的宏基因组测序,进行属水平和种水平物种富集分析,结果如图3、图4和图5所示,结果显示,与对照组相比,含乳双歧杆菌Probio-M8的酸奶显著提升了儿童特征性产乙酸菌Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacterium dentium、Bifidobacteriumhydrogenotrophica、Streptococcus agalactiae、Streptococcus dysgalactiae、Streptococcus thermophilus、Streptococcus urinalis和Streptococcus vestibularis的丰度,以上产乙酸菌丰度分别是对照组的4.99、1.25、2.16、1.37、3.80、1.34、20.54、10.50和1.92倍(图3);与对照组相比,含乳双歧杆菌Probio-M8酸奶显著提升了儿童特征性产丙酸菌Bifidobacterium adolescentis、Eubacterium hallii和Ruminococcus torques的丰度,提升倍数分别为4.03、1.71和1.70倍(图4);与对照组相比,含乳双歧杆菌Probio-M8酸奶显著提升了儿童特征性产丁酸菌Eubacterium hallii、Subdoligranulumvariabile、Ruminococcus torques和butyrate-producing bacterium、butyrate-producing bacterium SSC/2的丰度,以上产丁酸菌的丰度分别是对照组的1.71、1.44、1.70、1.39和1.42倍(图5)。
综上所述,本发明解析了中国儿童肠道菌群特点(与中国成人相比)及其特征性产SCFA菌群(Bifidobacterium、Streptococcus、Ruminococcus和Eubacterium等)。本发明提供的乳双歧杆菌组合物及其酸奶产品可以选择性富集中国儿童特征性产SCFA菌群,显著提升Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacteriumhydrogenotrophica、Streptococcus thermophilus等产乙酸代谢菌,提升Ruminococcustorques、Eubacterium hallii、Subdoligranulum variabile等产丁酸代谢菌,并提升Bifidobacterium adolescentis、Eubacterium hallii、Ruminococcus torques等产丙酸代谢菌。
实施例2乳双歧杆菌Probio-M8对儿童特征性产SCFA肠道菌的富集作用检测(2)
本实施例与实施例1的区别仅在于,干预组给予的含乳双歧杆菌Probio-M8的益生菌酸奶的各原料用量不同:
益生菌酸奶的原料由如下组分组成:菊粉(聚合度为2~10)0.7g,白砂糖7.5g,蛋白粉1.0g,乳双歧杆菌Probio-M8 0.025g(含乳双歧杆菌Probio-M8 5×109cfu),生牛乳补足至100g。
该益生菌酸奶产品中乳双歧杆菌Probio-M8的含量为5×107cfu/g酸奶。其制备方法与实施例1相同。
招募儿童志愿者共135名,招募标准为:年龄8-9岁的中国学龄儿童;一个月内未使用过益生菌、益生元制剂或抗生素,且入组时无呼吸道和胃肠道疾病症状。将志愿者分为2组,其中,对照组63人,每日正常膳食;干预组72人,除正常膳食外,每日饮用上述益生菌酸奶产品,每日饮用1次,每次服用量为200mL,干预周期为40天。
在干预前和干预结束后,分别采集志愿者的粪便样品,利用Illumina Miseq高通量测序平台进行粪便样品的宏基因组测序,通过属水平和种水平物种富集分析,获得肠道菌群数据。
实验结果显示,本实施例中的含乳双歧杆菌Probio-M8的儿童益生菌酸奶对于儿童特征性产SCFA肠道菌的靶向富集作用效果与实施例1基本相当。与对照组相比,含乳双歧杆菌Probio-M8的酸奶显著提升了儿童特征性产乙酸菌Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacterium dentium、Bifidobncterium pseudongum、Bifidobncterium longum、Bifidobacterium hydrogenotrophica、Streptococcusdysgalactiae、Streptococcus thermophilus、Streptococcus agalactiae、Streptococcus urinalis和Streptococcus vestibularis的丰度(图6),其中,Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacterium dentium、Bifidobncterium pseudongum、Bifidobncterium longum、Bifidobacteriumhydrogenotrophica、Streptococcus dysgalactiae、Streptococcus thermophilus、Streptococcus agalactiae、Streptococcus urinalis和Streptococcus vestibularis的丰度分别是对照组的29.2、8.6、12.6、15.5、11.9、16.9、38.8、89.0、22.4、40.5和19.2倍。
与对照组相比,含乳双歧杆菌Probio-M8酸奶显著提升了儿童特征性产丙酸菌Bifidobacterium adolescentis、Eubacterium hallii、Ruminococcus torques和Ruminococcus bromii的丰度,提升倍数分别为9.1、3.7、2.1和2.9倍(图7);与对照组相比,含乳双歧杆菌Probio-M8酸奶显著提升了儿童特征性产丁酸菌Eubacterium hallii、Subdoligranulum variabile、Ruminococcus torques、butyrate-producing bacterium、butyrate-producing bacterium SSC/2和Eubacterium dolichum的丰度,以上产丁酸菌的丰度分别是对照组的3.7、6.5、2.1、2.0、7.4和3.4倍(图8)。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (10)
1.乳双歧杆菌Probio-M8在制备用于靶向调控儿童特征产短链脂肪酸肠道菌的产品中的应用,所述乳双歧杆菌Probio-M8的保藏编号为CGMCC No.18610。
2.根据权利要求1所述的应用,其特征在于,所述儿童特征产短链脂肪酸肠道菌为产乙酸肠道菌、产丁酸肠道菌、产丙酸肠道菌中的一种或多种。
3.根据权利要求1或2所述的应用,其特征在于,所述儿童特征产短链脂肪酸肠道菌包括选自以下属细菌中的一种或多种:双歧杆菌(Bifidobacterium)、链球菌(Streptococcus)、瘤胃球菌(Ruminococcus)和真杆菌(Eubacterium);
优选地,所述儿童特征产短链脂肪酸肠道菌为选自Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacterium dentium、Bifidobacteriumhydrogenotrophica、Streptococcus agalactiae、Streptococcus dysgalactiae、Streptococcus thermophilus、Streptococcus urinalis、Streptococcus vestibularis、Bifidobncterium pseudongum、Bifidobncterium longum、Bifidobacteriumadolescentis、Ruminococcus torques、Ruminococcus bromii、Eubacterium hallii、Subdoligranulum variabile、butyrate-producing bacterium SSC/2、Eubacteriumdolichum中的一种或多种。
4.一种产品,其特征在于,包含乳双歧杆菌Probio-M8,所述乳双歧杆菌Probio-M8的保藏编号为CGMCC No.18610。
5.根据权利要求4所述的产品,其特征在于,所述产品为发酵乳制品,所述发酵乳制品的原料包含如下重量份的组分:乳原料80-95份,乳双歧杆菌Probio-M8 0.001-0.5份、108-1011cfu/100g原料。
6.根据权利要求4所述的产品,其特性在于,所述产品为调制乳粉,其由包括如下重量份的原料制备得到:生牛乳320-355份,脱脂乳粉25.8-28.6份,浓缩乳清蛋白粉0.48-0.55份,加糖炼乳23.5-26份,磷脂0.56-0.63份,大豆油2.9-3.3份,低聚果糖3.1-3.5份,低聚半乳糖0.73-0.82份,维生素预混料0.2-0.27份,(3R,3’R)-二羟基-β-胡萝卜素0.0089-0.01份,七水硫酸亚铁0.039-0.044份,七水硫酸锌0.029-0.033份,四水硫酸镁0.23-0.26份,碳酸钙0.94-1.04份,5%硫酸铜0.029-0.033份,富硒酵母0.0075-0.009份,氯化胆碱0.094-1.04份,5%叶黄素0.0043-0.0048份,酵母β-葡聚糖0.01-0.36份,二十二碳六烯酸0-0.65份,花生四烯酸0-1.3份,乳铁蛋白0-0.11份,磷脂酰丝氨酸0-1.0份,N-乙酰神经氨酸0-0.5份,异构化乳糖0-2.21份,柠檬酸0.01-0.43份,氢氧化钾0.001-0.104份,乳双歧杆菌Probio-M8 0.001-0.6份、5×108-5×1010cfu/100g原料;
所述维生素预混料包括:醋酸维生素A、维生素D3、DL-α-醋酸生育酚、植物甲萘醌、盐酸硫胺素、核黄素、盐酸吡哆醇、L-抗坏血酸、D-泛酸钙、D-生物素、叶酸、烟酰胺、氰钴胺和牛磺酸。
7.一种乳双歧杆菌组合物,其特征在于,其包括乳双歧杆菌Probio-M8和益生元,所述乳双歧杆菌Probio-M8的保藏编号为CGMCC No.18610;
所述益生元为选自菊粉、低聚半乳糖、低聚果糖、低聚异麦芽糖、低聚木糖、低聚甘露糖、半乳甘露聚糖、水苏糖、聚葡萄糖、抗性糊精中的一种或多种;
优选地,所述益生元为菊粉,所述菊粉的聚合度为2-10;
更优选地,所述组合物中,所述乳双歧杆菌Probio-M8与所述菊粉的比例为108-1011cfu:(0.2-2)g;优选为108-1011cfu:(0.5-1)g。
8.权利要求7所述的乳双歧杆菌组合物在食品、保健品或药物制备中的应用。
9.一种食品,其特征在于,所述食品包含权利要求7所述的乳双歧杆菌组合物,或者,所述食品的制备原料包含权利要求7所述的乳双歧杆菌组合物。
10.根据权利要求9所述的食品,其特征在于,所述食品为发酵乳制品,所述发酵乳制品的原料包含如下重量份的组分:乳原料80-95份,乳双歧杆菌Probio-M8 0.001-0.5份、108-1011cfu/100g原料,菊粉0.2-2份;
所述乳原料为选自生牛乳、复原乳中的一种或两种;
优选地,所述原料还包括糖类和蛋白粉;
更优选地,所述发酵乳制品的原料包含如下重量份的组分:乳原料80-95份,糖类3-15份,蛋白粉0.1-5份,乳双歧杆菌Probio-M8 0.001-0.05份、108-1011cfu/100g原料,菊粉0.2-2份。
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