CN117305158A - 一种具有改善记忆与认知功能的发酵乳乳清及其应用 - Google Patents
一种具有改善记忆与认知功能的发酵乳乳清及其应用 Download PDFInfo
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- CN117305158A CN117305158A CN202311155652.8A CN202311155652A CN117305158A CN 117305158 A CN117305158 A CN 117305158A CN 202311155652 A CN202311155652 A CN 202311155652A CN 117305158 A CN117305158 A CN 117305158A
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- lactobacillus helveticus
- ccfm1263
- fermented whey
- fermented
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Abstract
本发明公开了一种具有改善记忆与认知功能的发酵乳乳清及其应用,属于乳品加工与发酵技术领域。所述的发酵乳乳清以瑞士乳杆菌CCFM1263为发酵剂,发酵脱脂乳,并经沉淀、离心、膜过滤等操作除去蛋白后得到。瑞士乳杆菌CCFM1263发酵乳乳清能够改善记忆与认知障碍,有效缓解海马神经元的损伤,具有广泛的市场应用前景。
Description
技术领域
本发明涉及一种具有改善记忆与认知功能的发酵乳乳清及其应用,属于乳品加工与发酵技术领域。
背景技术
随着居民生活条件的改善,消费者对食品的消费需求也在不断升级,其中营养健康元素是多数人关注的重点。发酵乳由牛乳经乳酸菌发酵而成,是补充蛋白质和矿物质等营养物质的有效途径,然而目前市售发酵乳制品同质化严重问题,逐渐成为影响乳企发展的问题之一。而功能型乳制品的崛起改变了这一现状,由于其符合了消费者对健康生活方式的追求,受到强烈关注,成为乳品企业竞争中的热点。
“认知”指的是大脑对信息的获取、存储、提取和处理的所有活动和过程,包括记忆、学习、理解、判断、执行等多方面。认知功能障碍是指上述认知活动中的一项或多项受损,具体表现为一个人在记忆、学习新事物,集中注意力或作日常决策等方面存在困难。认知障碍从轻微到严重不等。轻度损伤时,人们可能逐渐开始注意到认知功能的变化,但仍然能够进行日常活动;待到严重损伤时,人们则会丧失理解事物的意义或重要性的能力,进而导致无法正常生活。Jia等人(Jia等,2020)的统计结果表明,在中国,截至2020年,60岁及以上的老年人中患认知障碍的概率约为15.5%,其中严重到痴呆程度的概率约为6.0%。《世界阿兹海默症报告2016》更是指出,2015年全球约有4680万痴呆症患者,预计到2050年,这一数字将增加至1.31亿。可见,随着人口老龄化的加剧,认知障碍已经成为一个全球性的公共卫生问题。同时随着生活压力和学习压力的增大,加上睡眠质量不佳,越来越多的年轻人也开始出现记忆力开下降。因此,如何改善记忆和认知功能越来越受到研究者的关注。
发酵乳乳清是在发酵乳制备过程中产生的副产品,具有多种营养成分,包括蛋白质、氨基酸、维生素、矿物质等,目前主要处理方式是直接作为动物饲料或者直接作为废水处理,导致额外的环境污染和费用,也构成了资源损失。为了有效利用发酵乳乳清,需要开发一种功能性的发酵乳清产品,但是发酵菌株的差异将影响发酵乳乳清的生物活性肽等功能成分及功能特性,现有技术中尚未发现能够发酵产生具有改善记忆与认知功能的发酵乳乳清的瑞士乳杆菌。因此,开发能产生功能性发酵乳乳清的瑞士乳杆菌,并开发更多有特色和功能特性的发酵乳乳清产品,以满足不同消费者的需求具有重要的意义。
发明内容
本发明提供一株瑞士乳杆菌(Lactobacillus helveticus)CCFM1263,所用菌株是从我国青海西宁地区采集的曲拉中筛选出的。于2022年5月17日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCC No:62417,保藏地址为广州市先烈中路100号大院59号楼5楼。
本发明还提供一种微生物制剂。
在一种实施方式中,所述微生物制剂中含有含有所述瑞士乳杆菌CCFM1263,或含有瑞士乳杆菌CCFM1263的发酵液,或含有瑞士乳杆菌CCFM1263的冻干粉,或含有瑞士乳杆菌CCFM1263灭活的菌体,或含有瑞士乳杆菌CCFM1263的裂解物,或含有瑞士乳杆菌CCFM1263的提取物。
在一种实施方式中,所述微生物制剂中含有瑞士乳杆菌的菌体数≥1×103CFU/ml或≥1×103CFU/g。
本发明还提供一种具有改善记忆与认知功能的的发酵乳乳清,所述发酵乳乳清由所述瑞士乳杆菌CCFM1263,或所述微生物制剂发酵制得。
本发明还提供瑞士乳杆菌CCFM1263,或所述微生物制剂所述的发酵乳乳清在制备辅助改善记忆的保健品中的应用。
本发明还提供瑞士乳杆菌CCFM1263,或所述微生物制剂所述的发酵乳乳清在制备改善记忆与认知的药品中的应用。
在一种实施方式中,所述药品还包含药物载体或药用辅料。
在一种实施方式中,所述药物载体包含微囊、微球、纳米粒和/或脂质体。
在一种实施方式中,所述药用辅料包含赋形剂和/或附加剂。
在一种实施方式中,所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
本发明还提供瑞士乳杆菌CCFM1263,或所述微生物制剂所述的发酵乳乳清在制备改善记忆与认知的食品中的应用。
在一种实施方式中,所述食品包括乳制品、豆制品或果蔬制品。
在一种实施方式中,所述改善记忆与认知包括但不限于如下方面:
(1)显著缩短水迷宫空间探索期第一次潜伏期;
(2)显著提高新物体识别能力;
(3)显著提高Y迷宫自发交替率;
(4)显著改善焦虑样行为;
(5)有效缓解海马神经元损伤。
本发明还提供制备所述发酵乳乳清的方法,以瑞士乳杆菌CCFM1263(原始编号为DQHXN-Q32M42)或所述微生物制剂为发酵剂,在30~42℃发酵脱脂乳12~72h,接着经沉淀、离心、膜过滤等操作除去蛋白得到所述发酵乳乳清。
在一种实施方式中,所述脱脂乳为将5~20%(w/w)的脱脂乳粉溶于水中配制得到。
在一种实施方式中,所述方法将已纯化的瑞士乳杆菌CCFM1263先后经MRS培养基和脱脂乳体系适应性培养,至菌浓度达108~109cfu/mL。
在一种实施方式中,所述方法将已活化的瑞士乳杆菌CCFM1263以(1~5)×108CFU/mL的浓度接种于脱脂乳中发酵,接种量为2%~4%(v/v)。
本发明还提供所述的瑞士乳杆菌,或所述的微生物制剂,或所述的发酵乳乳清在制备缓解海马组织损伤的产品中的应用。
有益效果:
1、本发明中应用的瑞士乳杆菌CCFM1263是可用于食品的菌株,安全健康,可用于制备各类食品、药品、微生物制剂。
2、本发明的瑞士乳杆菌CCFM1263发酵乳乳清可以显著增强乙酰胆碱酯酶的抑制性,与ATCC15009发酵乳乳清相比,乙酰胆碱酯酶的抑制性增强了456%;与GDMCC No.60811发酵乳乳清相比,乙酰胆碱酯酶的抑制性增强了77%;与脑活素相比,乙酰胆碱酯酶的抑制性增强了32%。
3、本发明的瑞士乳杆菌CCFM1263发酵乳乳清可以显著增强抗氧化能力,与GDMCCNo.60811发酵乳乳清相比,DPPH和ABTS自由基清除率分别增强了21%和17%。
4、本发明的瑞士乳杆菌CCFM1263发酵乳乳清可以显著改善东莨菪碱诱导的小鼠记忆与认知障碍,CCFM1263发酵乳乳清可以显著缩短水迷宫空间探索第一次到达平台期的178%,并提高了Y迷宫测试中显著提高自发交替率的15%。
5、本发明的瑞士乳杆菌CCFM1263发酵乳乳清可以有效缓解小鼠海马组织损伤。
生物材料保藏
本发明所提供的瑞士乳杆菌(Lactobacillus helveticus)CCFM1263,于2022年5月17日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCC No:62417,保藏地址为广州市先烈中路100号大院59号楼5楼。
附图说明
图1:瑞士乳杆菌(Lactobacillus helveticus)CCFM1263菌落形态图;
图2:瑞士乳杆菌发酵乳乳清对乙酰胆碱酯酶抑制能力和抗氧化能力评价。数据以平均数±标准差表示,n=3。柱形图上方不同字母a,b,c,d表示(p<0.05)差异有统计学意义。
图3:不同处理组小鼠水迷宫实验空间探索阶段代表性轨迹图;
图4:不同处理组小鼠水迷宫实验空间探索阶段第一次潜伏期;
图5:不同处理组小鼠新物体识别实验代表性轨迹图(左侧同心圆代表新物体,右侧同心圆代表旧物体)
图6:不同处理组小鼠Y迷宫自发交替率;
图7:不同处理组小鼠的海马组织形态(尼氏染色);
其中,*表示与模型组比较,*:p<0.05,**:p<0.01,***:p<0.001。
具体实施方式
技术术语:
本发明中的“接种量”是指加入种子液的体积与接种后培养液体积之比。
本发明中的“改善”是指在功能减退之后逆转、减轻、最小化、抑制、制止和/或停止功能减退进程的一种或多种临床症状。
本发明中“记忆和认知障碍”是指是与学习、记忆以及思维判断有关的大脑高级智能加工过程出现异常,例如,遗忘、记忆减退和记忆错误;所述记忆减退是指识记、保持、再认和回忆普遍减退。
本发明中“空间探索能力”是指是指探索物体形状和/或位置的能力。所述空间探索能力包括观察、思考、想象、认识和/或摸索物体形状和/或位置。
下述实施例中用于配制MRS液体/固体培养基的试剂购于国药集团化学试剂有限公司;下述实施例中涉及的脱脂乳粉购于光明乳业股份有限公司;下述实施例中涉及的SPF级6周龄雄性ICR小鼠购于维通利华实验动物有限公司。
下述实施例中涉及的培养基如下:
MRS固体培养基:蛋白胨10g/L、牛肉膏10g/L、葡萄糖20g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K2PO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO40.05 g/L、吐温80 1mL/L、琼脂17g/L。
MRS液体培养基:蛋白胨10g/L、牛肉膏10g/L、葡萄糖20g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K2PO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO40.05 g/L、吐温80 1mL/L。
实施例1:瑞士乳杆菌CCFM1263菌株分离鉴定方法
1、制备合适的样品稀释梯度并培养
取出-80℃冰箱中贮藏的青海西宁地区的曲拉,置于冰上缓慢解冻。震荡混匀后取0.5mL样品加入4.5mL无菌生理盐水中,完成一次10倍稀释,振荡混匀后再从该稀释液中取出0.5mL稀释液加入4.5mL无菌生理盐水中,完成第二次10倍稀释,以此类推,直至完成第五次10倍稀释(稀释到10-5),从每个梯度的稀释液中吸取100μL,均匀涂布于MRS固体培养基平板上,倒置于37℃条件下培养48h。
2、划线分离与纯化
取出长出菌落的平板,选取单菌落明显的梯度平板,挑取不同菌落形态的菌落,进行二次划线,直至纯化出所有单菌落。
3、菌种保藏
将纯化完成后每种菌株的单菌落挑入5mLMRS液体培养基中,置于37℃条件下培养24h,混匀后吸取1mL菌液至保菌管中,6000rpm离心3min,弃掉上清,加入1mL 30%(v/v)无菌甘油溶液,重悬,置于-80℃保存。
4、16S rDNA序列扩增
混匀后吸取1mL菌液6000rpm离心3min,倾去上清,无菌水洗两次,离心弃掉上清液,获得菌泥,以此为模板,进行PCR扩增。扩增体系共20μL,其中模板量为1μL,双向引物各0.5μL,Taq酶MasterMix为10μL,ddH2O为8μL。所用引物为27F:AGAGTT TGATCC TGG CCT CA(SEQ ID No 1),1492R:GGT TAC CTT GTTACGACT T(SEQ ID No 2)。扩增条件为:预变性,94℃,10min;变性,94℃,30s;退火,55℃,30s;延伸,72℃,2min;循环30次;最后72℃延伸10min;12℃保持10min。
5、测序与鉴定
将PCR样品送到苏州金唯智生物科技有限公司进行测序,得到菌株编号DQHXN-Q32M42的扩增核苷酸序列测序结果,将测序结果在美国国家生物技术信息中心(NCBI)序列数据库(http://www.ncbi.nlm.nih.gov/blast)中进行BLAST比对,可以确定本发明提供的菌株为瑞士乳杆菌(Lactobacillus helveticus)。
实施例2:瑞士乳杆菌CCFM1263发酵乳乳清的制备方法
将实施例1得到的瑞士乳杆菌CCFM1263接种于MRS液体培养基中,在37℃下培养18~24h,传代培养2~3次,至菌浓度达108~109cfu/mL。取出在MRS中活化后的菌液,6000×g离心5min,用11%(w/w)脱脂乳重悬至相同体积,按2%(v/v)的接种量接入11%(w/w)脱脂乳中,传代培养2~3次,至菌浓度达108~109cfu/mL。用Tris酸洗去蛋白,并调整活菌数至108CFU/mL~109CFU/mL,以2%~4%(v/v)的接种量接种于11%(w/w)脱脂乳中,30~42℃发酵12~72h,得到瑞士乳杆菌CCFM1263发酵乳。在瑞士乳杆菌CCFM1263发酵乳中加入1mol/L的氢氧化钠溶液调节样品pH至4.5~4.7后,于高压蒸汽灭菌锅中巴氏杀菌。将灭菌后的瑞士乳杆菌CCFM1263发酵乳于10000×g,4℃,离心10min,取上清液,再用0.45μm有机滤膜过滤,即得到瑞士乳杆菌CCFM1263发酵乳乳清。
实施例3瑞士乳杆菌CCFM1263发酵乳乳清乙酰胆碱酯酶抑制性和抗氧化能力1、乙酰胆碱酯酶的抑制性
依次将30μL碘代硫代乙酰胆碱、125μL 5,5'-二硫代双(2-硝基苯甲酸)(DTNB)、40μL4-羟乙基哌嗪乙磺酸(HEPES)以及50μL样品或脑活素加入到96孔板中,混匀后,在37℃下孵育15min。各组别的样品分别为:脑活素组(10mg/mL脑活素)、原乳组(11%(w/w)脱脂乳)、ATCC15009发酵乳乳清组(ATCC15009发酵乳乳清)、CCFM1263发酵乳乳清组(CCFM1263发酵乳乳清)和GDMCC No.60811发酵乳乳清组(GDMCC No.60811发酵乳乳清)。接着,加入30μL乙酰胆碱酯酶(AChE)(0.05U/mL)激活反应。15min后,用酶标仪测定412nm波长下各反应孔的吸光值。样品的AChE抑制活性计算公式为:AChE抑制活性(%)=(1-(A样品-A样品空白)/(A对照-A对照空白))×100。其中,用HEPES代替AChE作为样品空白组,用HEPES代替AChE和样品作为对照组,用HEPES代替样品作为对照空白组。分别检测各组的AChE抑制活性。其中ATCC15009和GDMCC No.60811发酵乳乳清的制备方法参照实施例2。
由图2A可知,CCFM1263发酵乳乳清组AChE抑制活性显著强于GDMCC No.60811发酵乳乳清组和脑活素组,ATCC15009发酵乳乳清组最弱。CCFM1263发酵乳乳清组比GDMCCNo.60811发酵乳乳清组增强了77%,比ATCC15009发酵乳乳清组增强了456%,比脑活素组增强了32%。
2、抗氧化能力测定
准确称取3.9432mg DPPH溶于10mL无水乙醇中,配制1mM DPPH工作液体。将100μL样品和100μLDPPH工作液加入到96孔板中,于室温下暗反应。30min后,用酶标仪测定510nm波长下各反应孔的吸光值。准确称取39.5mgABTS溶于10mL水,配制7.2mM ABTS溶液;称取6.8mg过硫酸钾溶于10mL水,配制2.5mM过硫酸钾溶液。将所得到的ABTS溶液和过硫酸钾溶液于暗处反应16h后得到ABTS工作液,备用。使用前需将ABTS工作液稀释至734nm处吸光度为0.8。将100μL样品和100μLABTS工作液加入到96孔板中,于室温下暗反应。15min后,用酶标仪测定734nm波长下各反应孔的吸光值。分别检测谷胱甘肽组(1mg/mL)、原乳组(11%(w/w)脱脂乳)、ATCC15009发酵乳乳清组、CCFM1263发酵乳乳清组和GDMCC No.60811发酵乳乳清组的DPPH和ABTS自由基清除率。
由图2B和C可知,CCFM1263发酵乳乳清组DPPH和ABTS自由基清除率显著强于GDMCCNo.60811发酵乳乳清组,ATCC15009发酵乳乳清组最弱。CCFM1263发酵乳乳清组DPPH和ABTS自由基清除率与GDMCC No.60811发酵乳乳清组比分别增强了21%和17%。
实施例4:瑞士乳杆菌CCFM1263发酵乳乳清对东莨菪碱诱导的认知障碍小鼠行为的影响
1、动物实验方案
取SPF级6周龄雄性ICR小鼠32只饲养于昼夜等分、恒温恒湿的屏障内。实验分为适应期、瑞士乳杆菌发酵乳乳清干预期及瑞士乳杆菌发酵乳乳清干预和东莨菪碱造模并行期。适应性喂养一周后,开始进入瑞士乳杆菌发酵乳乳清干预期,所有小鼠按照每组8只随机分成4组,分别为:空白组、模型组、ATCC15009发酵乳乳清组,GDMCC No.60811发酵乳乳清组和CCFM1263发酵乳乳清组,每日参照表1进行灌胃。其中,ATCC15009是瑞士乳杆菌模式菌株。灌胃第22天,开始进入瑞士乳杆菌发酵乳乳清干预和东莨菪碱造模并行期,先进行为期7天的水迷宫测试,休息5天后,进行历时2天的新物体识别实验。此时仍每日进行灌胃,灌胃30min后参照表1进行腹腔注射造模,若当日需进行行为学实验,行为学实验于腹腔注射30min后开始。具体的实验动物分组及处理方式见表1。
表1动物实验分组及处理方法
注:1mg/kg·BW的造模剂量中1mg为东莨菪碱含量,而非东莨菪碱溶液含量。
2、行为学评价指标与结果
Morris水迷宫用于测试实验小鼠的空间探索能力和学习记忆能力,具体来讲,Morris水迷宫所测试的是实验动物在多次训练中形成的稳定的空间位置认知。由于实验动物入水位置的变换,使得其难以根据自身位置,而是需要借助空间信息(外部线索提示)来定位隐蔽平台的位置,因此这是一种以他物为参照的参考认知,所形成的是一种空间参考记忆。实验装置由一个直径120cm、高60cm的水池和一个直径12cm的高度可调节、位置可移动的平台组成。正式实验期间,在水池中放入30cm深的水,并用黑色墨汁将水搅拌至深黑色,以平台在水下时不可见作为墨汁添加量的判断标准。水温一直保持在21℃左右。在水池的四周等距贴上四个不同颜色、不同形状的卡纸片作为标识物。实验主要包括三个部分:平台可见期(1天)、定位巡航期(第2~6天)和空间探索期(第7天)。三个阶段的测试时间均设置为60s,摄像头录像记录小鼠的游泳轨迹用于后续数字化分析。在定位巡航期,若小鼠在60s内未能找到平台,需将其引导至平台并在平台上停留15s以上;空间探索期则任小鼠自由探索60s。
由图3组复合热区图可知,空白组能够相对准确地记住平台所在位置,因而在其附近盘旋;而模型组则记忆出现了偏差或者没有记住平台所在位置,几乎未在平台附近停留,主要途径区域与平台相距甚远。对照组(ATCC15009发酵乳乳清组)的表现与模型组相似,极大地偏离了平台;而CCFM1263发酵乳乳清组和GDMCC No.60811发酵乳乳清组的实验结果与空白组相仿,能够在平台贴壁区域小范围盘旋。此外,从代表性轨迹图可见,空白组采取的探索策略为直线式,而模型组小鼠的游走路径则毫无规律,属于边缘式和随机式。相较于模型组,ATCC15009发酵乳乳清组在探索策略上有所进步,属于趋向式,CCFM1263发酵乳乳清组比GDMCC No.60811发酵乳乳清组的表现则更佳,接近空白组,属于直线式。
图4的结果也表明东莨菪碱造模引起了小鼠的记忆与认知障碍,具体表现为与空白组相比,模型组小鼠水迷宫空间探索期第一次到达平台的时间(即第一次潜伏期)显著延长。与模型组相比,GDMCC No.60811发酵乳乳清和CCFM1263发酵乳乳清能显著缩短小鼠水迷宫空间探索期第一次潜伏期,分别缩短了160%和178%,且显著低于ATCC15009发酵乳乳清组。
新物体识别测试旨在根据动物探索新物体的天性来评估其学习和记忆能力。实验历时两天,包括适应期、熟悉期和测试期。第一天为适应期,小鼠在40cm×40cm×30cm的黑色旷场箱内自由探索,时长为6min。次日上午为熟悉期,黑色旷场箱内被放入两个一模一样的六棱柱木块,木块距离两边的墙面均为10cm。小鼠在其中自由探索6min。待小鼠回到原笼盒休息6h后,开始进行测试期实验。此时将其中一个六棱柱木块替换为圆锥体木块,再次让小鼠自由探索。红外摄像头录像记录小鼠6min内的探索轨迹。每次实验结束后需用75%的酒精擦拭Y迷宫以消除气味。
由图5可知,在新物体测试阶段,与空白组相比,模型组小鼠更倾向于贴壁行走,表现出更高的焦虑水平,对新物体的探索倾向也不明显。ATCC15009发酵乳乳清组小鼠的表现与模型组相似;而GDMCC No.60811发酵乳乳清组和CCFM1263发酵乳乳清组小鼠焦虑样水平明显改善,中间区域的探索痕迹明显,且相较于右侧旧物体,更倾向于探索左侧新物体区域,整体表现趋向于模型组。
自发交替是一种衡量空间工作记忆的标准,它由啮齿动物对未知区域的好奇心驱动,可以通过让小鼠探索Y迷宫的所有三个臂来进行评估。工作记忆完好的小鼠,会记住之前访问过的臂,进而表现为交替进入三个臂,图6的结果显示,与模型组相比,仅CCFM1263发酵乳乳清组在Y迷宫测试中显著提高自发交替率,并提高了15%,提示CCFM1263的发酵乳乳清可显著改善短期记忆。
综上所述,行为学(Morris水迷宫空间探索期,新物体识别测试期和Y迷宫自发交替期)实验结果表明,CCFM1263发酵乳乳清能够有效改善东莨菪碱诱导的记忆和认知障碍。实施例5:瑞士乳杆菌CCFM1263发酵乳乳清对东莨菪碱诱导的认知障碍小鼠海马组织形态的影响
按照实施例4的动物实验设计,在瑞士乳杆菌发酵乳乳清干预和东莨菪碱造模并行期结束的次日,牺牲小鼠。具体操作为麻醉小鼠并迅速摘眼球取血,辅以颈椎脱臼法处死。对处死后小鼠进行解剖,摘取全脑,于4%多聚甲醛固定液中固定。固定48h后进行石蜡包埋、切片,采用尼氏染色观察海马组织形态。
图7尼氏染色结果显示,空白组小鼠海马CA1区和DG区尼氏体染色均匀,胞体形状规则;而模型组小鼠海马CA1区和DG区完整存活的神经元密度低,排列散乱,染色过深,形态异常,出现核固缩,呈菱形和三角形。CCFM1263发酵乳乳清组神经元细胞形态规整、核质分界清晰,深染现象明显缓解,能有效缓解这种损伤;ATCC15009发酵乳乳清组的损伤情况虽较模型组略有减轻,但损伤仍是很严重。以上结果说明CCFM1263发酵乳乳清能够缓解东莨菪碱引起的小鼠海马神经元损伤,进而改善认知障碍。
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 (15)
1.一株瑞士乳杆菌(Lactobacillus helveticus)CCFM1263,其特征在于,所述瑞士乳杆菌CCFM1263已于2022年5月17日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCCNo:62417。
2.一种微生物制剂,其特征在于,所述微生物制剂中,含有权利要求1所述瑞士乳杆菌CCFM1263,或含有瑞士乳杆菌CCFM1263的发酵液,或含有瑞士乳杆菌CCFM1263的冻干粉,或含有瑞士乳杆菌CCFM1263灭活的菌体,或含有瑞士乳杆菌CCFM1263的裂解物,或含有瑞士乳杆菌CCFM1263的提取物。
3.根据权利要求2所述的微生物制剂,其特征在于,所述含有瑞士乳杆菌CCFM1263,或含有瑞士乳杆菌CCFM1263的冻干粉,或含有瑞士乳杆菌CCFM1263灭活的菌体的微生物制剂中,菌体数≥1×103CFU/ml或≥1×103CFU/g。
4.一种改善记忆与认知功能的发酵乳乳清,其特征在于,所述发酵乳乳清由权利要求1所述瑞士乳杆菌CCFM1263,或权利要求2或3所述微生物制剂发酵制得。
5.权利要求1所述的瑞士乳杆菌,或权利要求2或3所述的微生物制剂,或权利要求4所述的发酵乳乳清在制备辅助改善记忆的保健品中的应用。
6.权利要求1所述的瑞士乳杆菌,或权利要求2或3所述的微生物制剂,或权利要求4所述的发酵乳乳清在制备改善记忆与认知的药品中的应用。
7.根据权利要求6所述的应用,其特征在于,所述药品还包含药物载体或药用辅料。
8.根据权利要求7所述的应用,其特征在于,所述药物载体包含微囊、微球、纳米粒和/或脂质体。
9.根据权利要求8所述的应用,其特征在于,所述药用辅料包含赋形剂和/或附加剂。
10.根据权利要求9所述的应用,其特征在于,所述药品的剂型为粉剂、颗粒剂、胶囊剂、片剂、丸剂或口服液。
11.权利要求1所述的瑞士乳杆菌,或权利要求2或3所述的微生物制剂,或权利要求4所述的发酵乳乳清在制备改善记忆与认知的食品中的应用。
12.根据权利要求11所述的应用,其特征在于,所述食品包括乳制品、豆制品或果蔬制品。
13.制备权利要求4所述的发酵乳乳清的方法,其特征在于,以权利要求1所述瑞士乳杆菌CCFM1263,或权利要求2或3所述的微生物制剂为发酵剂,于30~42℃发酵脱脂乳12~72h,并除去蛋白。
14.根据权利要求13所述的方法,其特征在于,所述发酵剂的接种量按体积比计,为2%~4%,所述发酵剂中瑞士乳杆菌CCFM1263的浓度为(1~5)×108cfu/mL。
15.权利要求1所述的瑞士乳杆菌CCFM1263,或权利要求2或3所述的微生物制剂,或权利要求4所述的发酵乳乳清在制备缓解海马组织损伤的产品中的应用。
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