CN115624517A - 一种益生菌后生元制品及其在促进皮肤健康中的应用 - Google Patents
一种益生菌后生元制品及其在促进皮肤健康中的应用 Download PDFInfo
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Abstract
本发明公开了一种益生菌后生元制品及其在促进皮肤健康中的应用，其中，一种益生菌后生元制品，包括：益生菌经发酵后的灭活菌体及其代谢产物；所述益生菌至少包括副干酪乳杆菌Probiosci‑101和发酵乳杆菌ProSci‑602；所述副干酪乳杆菌Probiosci‑101的保藏编号为CGMCCNo.25085，所述发酵乳杆菌ProSci‑602的保藏编号为CGMCC No.25451。本发明通过2种菌株的相互配合，可以获得能更有效改善皮肤状态的代谢产物，效果显著。
Description
技术领域
本发明涉及生物技术领域，具体涉及一种益生菌后生元制品及其在促进皮肤健康中的应用。
背景技术
皮肤作为人体最大的器官，皮肤上长驻着大约一千亿个微生物，也就是说大约一毛钱硬币大小的一片皮肤上，生存着超过两百万个微生物。这些微生物中大约有74-80％是细菌，5-10％是真菌，还有10-20％是噬菌体(细菌的天敌)，不同的物种之间维持着微妙的平衡。这种平衡不仅因人而异，如同基因和指纹一样，世界上找不到两个菌落分布相同的人，这个平衡还时时刻刻受着环境，气候，年龄，以及生活***衡，我们皮肤表面的微生态时时刻刻经受着各种挑战，研究显示：皮肤的微生态容易受到环境污染和年龄增长的影响。环境污染有可能引入外来微生物，使得皮肤微生态变得越来越复杂，失去平衡；年龄增长带来的皮脂分泌下降，尤其是更年期前后，会改变皮肤以皮脂为食的微生物比例下降，改变平衡态的比例。皮肤微生态失衡，最常见的后果就是痤疮，皮肤过敏，以及干燥脱皮等等。
后生元为直接在体外培养益生菌代谢后的产物，可以直接对皮肤产生正面作用，也是如今护肤品中最常见的微生态技术手段。有调查数据显示，听说且购买过“益生菌”成分护肤品的女性比例高达66％，说明采用益生菌后生元已普遍被接受。但现有公开的益生菌后生元产品，其代谢产物虽然能达到一定的维持皮肤微生态平衡、调节油水平衡，进而改善皮肤状态的目的，但是，其改善效果并不十分明显，尤其是在改善皮肤黑色素和皮肤皱纹上，效果并不理想。
发明内容
因此，本发明要解决的技术问题在于克服现有技术中公开的益生菌后生元产品对皮肤的改善效果并不十分明显的缺陷，从而提供一种能更有效改善皮肤状态的一种益生菌后生元制品及其在促进皮肤健康中的应用。
一种益生菌后生元制品，包括：益生菌经发酵后的灭活菌体及其代谢产物；所述益生菌至少包括副干酪乳杆菌Probiosci-101和发酵乳杆菌 ProSci-602；所述副干酪乳杆菌Probiosci-101的保藏编号为CGMCC No.25085，所述发酵乳杆菌ProSci-602的保藏编号为CGMCC No.25451。
本发明产品中包含的副干酪乳杆菌Probiosci-101，分离自新疆帕米尔高原酸牛乳中，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏号为CGMCC No.25085，分类命名为副干酪乳杆菌Lactobacillus paracasei，保藏时间为2022年6月13日，保藏地址为北京市朝阳区北辰西路1号院3号，经保藏中心于2022年6月13日检测为存活菌株。该菌株具有良好的抵抗胃酸、胆盐环境的能力；可改善脂质代谢，降低血脂，平衡机体的免疫功能。
本发明中产品中包含的植物乳杆菌发酵乳杆菌ProSci-602，分离自健康妇女母乳中，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏号为CGMCC No.25451，分类命名为发酵乳杆菌Lactobacillus fermentum，保藏时间为2022年8月1日，保藏地址为北京市朝阳区北辰西路1号院3号，经保藏中心于2022年8月1日检测为存活菌株。试验研究证明：发酵乳杆菌ProSci-602具有良好的耐受胃肠液、胆盐的能力，还可以阻止龋齿菌等口腔中潜在致病菌的定植和感染。
所述代谢产物中包含50mg/g以上的有机酸，7mg/g以上的短链脂肪酸， 2mg/g以上的氨基酸，90mg/g以上的小分子多肽，7mg/g以上具有抑菌功效的活性物质。优选的，包含100mg/g以上的小分子多肽。
所述有机酸包括柠檬酸、琥珀酸、酒石酸、苹果酸、4-羟基苯乳酸、苯丙氨酸、苯甲酸和草酸中的一种或多种；
所述短链脂肪酸包括乳酸、丙酸、丁酸、己酸和乙酸中的一种或多种；
所述氨基酸包括缬氨酸、酪氨酸、脯氨酸、苯丙氨酸、亮氨酸、异亮氨酸、丙氨酸、冬氨酸、谷氨酸、赖氨酸和组氨酸中的一种或多种；
所述活性物质包括：花生酸、甲氧基查耳酮、他克莫司、罗他霉素、异硬脂酸、马诺利德、咖啡酸、松脂素、青黛酮、神经酰胺、虾青素、赤霉素、五味子乙素、麦角甾醇、泰乐菌素、维生B2、大黄酸、柠檬黄素、脱皮甾酮、赤藓酮糖、茶碱、肉桂酸苄酯、丹酚酸、氢化可的松、角鲨胺、胡莫柳酯、荜茇明宁碱和维生素A酸中的一种或多种。
所述柠檬酸的含量为50mg/g以上；
所述乳酸的含量为5mg/g以上，所述己酸的含量为1.5mg/g以上；
所述缬氨酸的含量为0.6mg/g以上，酪氨酸的含量为0.4mg/g以上，脯氨酸的含量为0.3mg/g以上，苯丙氨酸的含量为0.2mg/g以上；
所述小分子多肽中二肽的含量为30mg/g以上，三肽的含量为50mg/g 以上，四肽的含量为10mg/g以上，五肽的含量为0.5mg/g以上；
所述花生酸的含量为2mg/g以上，甲氧基查耳酮的含量为0.9mg/g以上，他克莫司的含量为0.8mg/g以上，罗他霉素的含量为0.8mg/g以上，异硬脂酸的含量为0.8mg/g以上，马诺利德的含量为0.6mg/g以上，咖啡酸的含量为0.2mg/g以上，松脂素的含量为0.1mg/g以上。
所述副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602的菌落数目之比为2-3:2-3；
优选的，所述副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602的菌落数目之比为1:1或2:3。
所述发酵所用的培养基按质量百分数计，其组成包括：全脂豆粉 1.5％-7％、脱脂乳粉11％-18％、柠檬酸钠0.05％-0.5％，余量为水。
所述制品为化妆品，优选为功能性化妆品。
一种益生菌后生元制品的制备方法，包括：
获取上述的一种益生菌后生元制品中的益生菌菌种和培养基；
将益生菌菌种接种到灭菌后的培养基中，恒温发酵至pH为4.5-4.6停止，灭活并干燥后获得益生菌后生元制品。
所述恒温发酵的温度为33-37℃。
所述培养基在配置完成之后以及在灭菌之前还包括均质的步骤；
优选的，均质的温度为55-60℃、压力为18～20.0Mpa；
所述培养基灭菌时的温度为95℃，灭菌的时间为60min；
所述发酵之后、灭活之前还包括均质的步骤，均质的温度为55-60℃、一级压力为18～20.0Mpa、二级压力为5.0Mpa；
所述干燥为喷雾干燥。
上述的一种益生菌后生元制品或上述的制备方法制备得到的一种益生菌后生元制品在促进皮肤健康中的应用。
所述促进皮肤健康的应用包括但不限于如下任意一种中的应用：
1)辅助预防和/或改善面部皮肤状态；
2)制备辅助预防和/或改善面部皮肤状态的产品中的应用；
所述皮肤状态包括但不限于增加面部弹性、减少面部皱纹、增加面部水分含量、减少面部油性、减少面部黑色素。
本发明技术方案，具有如下优点：
本发明提供的一种益生菌后生元制品，优化了发酵用的益生菌菌株，选择不同来源、不同种类和不同益生特性的益生菌菌株相互配合，本发明中的益生菌菌株至少包括副干酪乳杆菌Probiosci-101和发酵乳杆菌 ProSci-602；通过上述至少2种菌株的相互配合，可以更加有效地将培养基底物转化为更加有利于改善皮肤状态的代谢产物，提高益生菌后生元制品在改善皮肤健康中的应用效果，尤其是不仅仅能达到常规调整皮肤上微生物菌群的目的，达到水油平衡，并且还能有效达到增加面部弹性、减少面部皱纹、减少面部黑色素等效果，综合效果十分优异。
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为实验例2中志愿者使用益生菌后生元28天后，皮肤含水量水平；图a：使用后生元护肤品0天和28天；图b：使用含和不含后生元护肤品28天；
图2为实验例2中志愿者使用益生菌后生元28天后，皮肤弹性情况；图a：使用后生元护肤品0天和28天；图b：使用含和不含后生元护肤品28天；
图3为实验例2中志愿者使用益生菌后生元28天后，皮肤黑色素水平；图a：使用后生元护肤品0天和28天；图b：使用含和不含后生元护肤品28天；
图4为实验例2中志愿者使用益生菌后生元28天后，皮肤油性水平；图a：使用后生元护肤品0天和28天；图b：使用含和不含后生元护肤品28天；
图5为实验例2中志愿者使用益生菌后生元28天后的皱纹水平；图a：使用后生元护肤品0天；图b：使用后生元护肤品28天；图c：使用后生元护肤品0天和28天；图d：使用含和不含后生元护肤品28天。
具体实施方式
实施例1
一种益生菌后生元制品，其制备方法如下：
1)培养基称料：将全脂豆粉5％、脱脂乳粉15％、柠檬酸钠0.25％和水 79.75％按比例需求混合；
2)化料：58℃化料，得到料液；
3)均质：58℃，压力为19.0Mpa；
4)杀菌：上述3)均质后的料液进行95℃、60min杀菌；
5)冷却：杀菌后的料液冷却至35℃；
6)添加菌种和乳糖酶：料液中加入活菌数量比为1:1的菌种，副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602在料液中菌种的接种总量为 4×106CFU/mL。按生产需求添加0.5g/1000g的乳糖酶，可通过酶解为益生菌提供碳源；
7)恒温发酵：35℃进行恒温发酵，至pH4.5-4.6停止，此发酵步骤大约需要18-19h，此实施例中发酵18h，发酵终点的pH值为4.6；
8)灭活：按生产需求混合麦芽糊精(为喷雾干燥提供理想的液体分散)，预热至58℃，经一级压力19.0Mpa、二级压力5.0Mpa均质后，进行杀菌灭活，杀菌灭活的参数条件为75℃、15min；
9)喷雾干燥：将灭活的菌悬液经喷雾干燥(进风温度160℃，出风温度 45℃)得到益生菌后生元粉。
实施例2
一种益生菌后生元制品，本实施例与实施例1的区别在于，培养基组成不同，本实施例中培养基组成为：全脂豆粉1.5％、脱脂乳粉18％、柠檬酸钠0.05％，余量为水。
步骤2)中的化料温度为55℃；
步骤3)中的均质温度为55℃，压力为20.0Mpa；
步骤7)中的发酵时间为19h，发酵终点的pH为4.5；
步骤8)中的均质温度为60℃，一级压力为18.0Mpa。
实施例3
一种益生菌后生元制品，本实施例与实施例1的区别在于，培养基组成不同，本实施例中培养基组成为：全脂豆粉7％、脱脂乳粉11％、柠檬酸钠0.5％，余量为水。
步骤2)中的化料温度为60℃；
步骤3)中的均质温度为60℃，压力为18Mpa；
步骤7)中的发酵时间为19h，发酵终点的pH为4.5；
步骤8)中的均质温度为55℃，一级压力为20.0Mpa。
实施例4
一种益生菌后生元制品，本实施例与实施例1的区别在于，副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602的菌落数目之比为2:3。
实施例5
一种益生菌后生元制品，本实施例与实施例1的区别在于，副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602的菌落数目之比为3:2。
试验例1
将实施例1-5制备得到的益生菌后生元粉分别采用LC-MS测定益生菌后生元中含有的代谢产物。因各成分的检测手段为常规手段，具体检测过程在此不再赘述。
实施例1-5得到的益生菌后生元粉测定结果如下表1-3所示。表1为实施例1-5中益生菌后生元粉中有机酸和短链脂肪酸的测定结果；表2为实施例1-5中益生菌后生元粉中氨基酸和小分子多肽的测定结果，表3为实施例 1-5中益生菌后生元粉中活性物质的测定结果。
表1
表2
表3
由上表可知实施例2-5检测得到的各成分的含量与实施例1无显著性差异。
试验例2
验证实施例1制备得到的益生菌后生元粉对皮肤健康的改善作用，具体的：
1试验分组
随机选择110名(女性60名，男性50名)无皮肤疾病，无皮肤感染的受试者，年龄在25-28岁之间。所有志愿者在前1周(第0周至第1周) 控制不使用其他美容产品。随机分为后生元组和对照组。在第2周至第5 周，分别使用提供的美肤产品(后生元组：添加1％益生菌后生元粉的睡眠面膜膏组；对照组：不加后生元面膜膏组)。在采样前的8小时内，受试者被要求不要用任何东西(包括水)清洗，其他情况下则允许进行典型的日常卫生操作。两组产品均为免洗式睡眠面膜膏，产品使用方法为：
(1)晚间洁面时，将毛孔中的油污及化妆品洗净；
(2)取适量本产品，轻薄均匀涂抹于脸部，避开眼周区域；
(3)涂抹全脸后，静待20分钟，轻柔按摩至吸收；
(4)吸收后，可免洗直接入睡或者清洗后入睡。
0、28天分别使用肤质诊断专家Derma-Expert MC760，对受试者面部皮肤表型(皮肤水分含量、皮肤弹性、皮肤黑色素、皮肤油性、皮肤皱纹) 进行检测。
2实验结果
气相色谱检测结果显示：后生元组在皮肤水分含量、皮肤弹性、皮肤黑色素、皮肤油性、皮肤皱纹改善上都显著优于对照组。试验结果如图1-5 所示，具体的：
皮肤水分含量检测指标分析结果如图1所示，后生元组和对照组的皮肤水分含量水平在治疗后均有上升的趋势，但后生元组上升更显著 (p<0.01)。
皮肤弹性检测指标分析结果如图2所示，后生元组和对照组在皮肤弹性方面在治疗后均有上升的趋势，但后生元组上升更显著(p<0.01)。
皮肤黑色素检测指标分析结果如图3所示，后生元组在降低皮肤黑色素水平上效果显著(p<0.01)，对照组在治疗28天后黑色素水平上升。
皮肤油性检测指标分析结果如图4所示，后生元组和对照组在皮肤油性方面在治疗后均有下降的趋势，但后生元组下降更显著(p<0.01)。
皮肤皱纹检测指标分析结果如图5所示，后生元组和对照组在皮肤皱纹方面在治疗后均有下降的趋势，但后生元组下降更显著(p<0.01)。
3结论
通过对受试者面部皮肤水平检测分析发现，后生元可以通过降低皮肤黑色素、油性和皱纹水平(p<0.01)，并且增加皮肤弹性和皮肤含水量 (p<0.01)来达到修护皮肤的效果。经过人体试验表明，该产品对于修护皮肤有显著的功效。
显然，上述实施例仅仅是为清楚地说明所作的举例，而并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (10)
1.益生菌后生元制品，其特征在于，包括：益生菌经发酵后的灭活菌体及其代谢产物；所述益生菌至少包含副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602；所述副干酪乳杆菌Probiosci-101的保藏编号为CGMCC No.25085，所述发酵乳杆菌ProSci-602的保藏编号为CGMCC No.25451。
2.根据权利要求1所述的益生菌后生元制品，其特征在于，所述代谢产物中包含50mg/g以上的有机酸，6mg/g以上的短链脂肪酸，2mg/g以上的氨基酸，90mg/g以上的小分子多肽，7mg/g以上的活性物质。
3.根据权利要求2所述的益生菌后生元制品，其特征在于，所述有机酸包括柠檬酸、琥珀酸、酒石酸、苹果酸、4-羟基苯乳酸、苯丙氨酸、苯甲酸和草酸中的一种或多种；
所述短链脂肪酸包括乳酸、丙酸、丁酸、己酸和乙酸中的一种或多种；
所述氨基酸包括缬氨酸、酪氨酸、脯氨酸、苯丙氨酸、亮氨酸、异亮氨酸、丙氨酸、冬氨酸、谷氨酸、赖氨酸和组氨酸中的一种或多种；
所述活性物质包括：花生酸、甲氧基查耳酮、他克莫司、罗他霉素、异硬脂酸、马诺利德、咖啡酸、松脂素、青黛酮、神经酰胺、虾青素、赤霉素、五味子乙素、麦角甾醇、泰乐菌素、维生B2、大黄酸、柠檬黄素、脱皮甾酮、赤藓酮糖、茶碱、肉桂酸苄酯、丹酚酸、氢化可的松、角鲨胺、胡莫柳酯、荜茇明宁碱和维生素A酸中的一种或多种。
4.根据权利要求3所述的益生菌后生元制品，其特征在于，
所述柠檬酸的含量为50mg/g以上；
所述乳酸的含量为5mg/g以上，所述己酸的含量为1.5mg/g以上；
所述缬氨酸的含量为0.6mg/g以上，酪氨酸的含量为0.4mg/g以上，脯氨酸的含量为0.2mg/g以上，苯丙氨酸的含量为0.2mg/g以上；
所述小分子多肽中二肽的含量为30mg/g以上，三肽的含量为50mg/g以上，四肽的含量为10mg/g以上，五肽的含量为0.6mg/g以上；
所述花生酸的含量为2mg/g以上，甲氧基查耳酮的含量为0.9mg/g以上，他克莫司的含量为0.8mg/g以上，罗他霉素的含量为0.8mg/g以上，异硬脂酸的含量为0.8mg/g以上，马诺利德的含量为0.6mg/g以上，咖啡酸的含量为0.2mg/g以上，松脂素的含量为0.1mg/g以上。
5.根据权利要求1-4任一项所述的益生菌后生元制品，其特征在于，所述副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602的菌落数目之比为2-3:2-3；
优选的，所述副干酪乳杆菌Probiosci-101和发酵乳杆菌ProSci-602的菌落数目之比为1:1或2:3。
6.根据权利要求1-5任一项所述的益生菌后生元制品，其特征在于，所述发酵所用的培养基按质量百分数计，其组成包括：全脂豆粉1.5％-7％、脱脂乳粉11％-18％、柠檬酸钠0.05％-0.5％，余量为水。
7.根据权利要求1-6任一项所述的益生菌后生元制品，其特征在于，所述制品为化妆品，优选为功能性化妆品。
8.根据权利要求1-7任一项所述的益生菌后生元制品的制备方法，其特征在于，包括：将益生菌菌种接种到灭菌后的培养基中，恒温发酵至pH为4.5-4.6停止，灭活并干燥后获得益生菌后生元制品。
9.权利要求1-7任一项所述的益生菌后生元制品以及权利要求8的制备方法制备得到的益生菌后生元制品在促进皮肤健康中的应用。
10.根据权利要求9所述的应用，其特征在于，所述促进皮肤健康的应用包括但不限于如下任意一种中的应用：
1)辅助预防和/或改善面部皮肤状态；
2)制备辅助预防和/或改善面部皮肤状态的产品中的应用；
所述皮肤状态包括但不限于增加面部弹性、减少面部皱纹、增加面部水分含量、减少面部油性或减少面部黑色素。
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