CN112725219B - 一种青春双歧杆菌菌株及其应用 - Google Patents
一种青春双歧杆菌菌株及其应用 Download PDFInfo
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- CN112725219B CN112725219B CN202011420948.4A CN202011420948A CN112725219B CN 112725219 B CN112725219 B CN 112725219B CN 202011420948 A CN202011420948 A CN 202011420948A CN 112725219 B CN112725219 B CN 112725219B
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- bifidobacterium adolescentis
- bifidobacterium
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Abstract
本发明公开了一种青春双歧杆菌菌株ZJ2,该菌株是从健康百岁长寿老人粪便中分离筛选而得,该菌株对酸和胆盐具有良好的耐受作用,培养原液以及5倍浓缩液对小鼠无毒性作用,青春双歧杆菌ZJ2对外源加入的0.1mg/ml胆固醇孵育48h后胆固醇的降解率29.15%,对大肠杆菌ATCC33849、沙门氏菌ATCC14028、绿脓杆菌ATCC9027、肺炎克雷伯杆菌ATCC700603、金黄色葡萄球菌NCTC8325、单增李斯特菌ATCC19115均有一定的抑制效果以及对菌株ZJ2对万古霉素(Van)、红霉素(Ery)以及四环素(Tet)敏感,同时青春双歧杆菌菌株ZJ2具有良好的抗氧化能力。
Description
技术领域
本发明属于益生菌技术领域,具体涉及一种青春双歧杆菌菌株。
背景技术
肠道作为人体最庞大、最复杂的微生态系统,在维持机体正常免疫防御功能中发挥着极其重要的作用,肠道菌群本身及其代谢产物越来越受到科学家们的重视。人体肠道中定植着大量的微生物。肠道微生物与人体健康与疾病之间存在着十分密切的关系。肠道微生物可分为有益菌、中性菌和有害菌3 大类。有益菌,主要以各种双歧杆菌、乳酸杆菌为代表的益生菌群体,是人体健康不可缺少的要素,可以合成各种维生素,发挥着参与食物的消化,促进肠道蠕动,抑制致病菌群的生长,分解有害、有毒物质等重要作用。有害菌,即能够产生致癌物等有害物质,或者影响免疫系统的功能的致病菌。中性菌,即具有双重作用的细菌,如大肠杆菌、肠球菌等,在正常情况下对健康有益,一旦肠道内的益生菌屏障受到破坏,肠道通透性增加,中性菌就会加入有害菌的大军,威胁人体健康。
益生菌是一类可以改善人体肠内微生态的平衡,并对机体有正面效益的活性微生物,能够定植于人体肠道、生殖系统内,具有缓解不耐乳糖症状、预防生殖系统感染、增强人体免疫力、降胆固醇、帮助吸收营养成分、增强植物新陈代谢等有益作用。人体内常见的益生菌主要有嗜酸乳杆菌、双歧杆菌、乳酸菌、酪酸梭菌、酵母菌和放线菌等,其中双歧杆菌是一种专性厌氧的革兰氏阳性细菌,不运动,形态多变,呈现弯曲杆状、L、V或Y形等多种形态。细胞呈杆状,其末端分叉,故命名为双歧杆菌。
双歧杆菌主要寄居在人体的胃肠道内,分布在胃肠的数量及种类会随年龄阶段的增长而减少,分布最多的是母乳婴儿时期,是健康者肠道内的优势菌种,其种类和数量随年龄呈动态变化,而且其改变与机体内许多生理病理变化关系密切。随着年龄增加,有害菌的数量增多,产生的毒素加速肠道老化和免疫力下降等因素,正常老年人肠道有益菌(如双歧杆菌)的丰度降低。然而有文章报道发现健康百岁老人肠道内的双歧杆菌种类丰富,研究者对这些百岁老人进行体检发现:他们体内的双歧杆菌等有益菌占比率明显高于普通老人,相当于青少年水平。通过多年考察,研究者们认为这些百岁老人与其所在的良好的自然生态环境和生活习惯密不可分。
发明内容
本发明的目的在于提供一种青春双歧杆菌菌株,克服了现有技术的不足,在长寿老人的粪便中分离筛选出了青春双歧杆菌菌株ZJ2,并对该菌株的环境耐受情况、抑菌作用、自由基的清除能力和还原力大小进行试验。
为解决上述问题,本发明所采取的技术方案如下:
一种青春双歧杆菌菌株,所述菌株由健康百岁长寿老人的粪便中分离筛选得到的,命名为青春双歧杆菌(Bifidobacterium adolescents)ZJ2,且已于 2019年11月04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,保藏编号为CGMCC NO.18901。
本发明还保护了一种活性成分中含有上述青春双歧杆菌菌株ZJ2的菌剂。
本发明还保护了一种成分中含有上述青春双歧杆菌菌株ZJ2的菌剂的组合物。
本发明还保护了上述组合物或者菌剂在食品中的应用。
本发明还保护了上述组合物或者菌剂在保健产品中的应用。
本发明最后还保护了上述组合物或者菌剂在抑制胃肠道致病菌药物中的应用。
青春双歧杆菌ZJ2菌株在pH 3.0环境中孵育2h后仍能存活,对0.3%胆盐环境也有一定的耐受;对青春双歧杆菌ZJ2对大肠杆菌ATCC33849、沙门氏菌ATCC14028、绿脓杆菌ATCC9027、肺炎克雷伯杆菌ATCC700603、金黄色葡萄球菌NCTC8325、单增李斯特菌ATCC19115均有抑制效果;对外源加入0.1mg/ml胆固醇孵育48h后胆固醇的降解率为29.15%;且对DPPH自由基和ABTS+自由基有良好的清除能力,有良好的还原力并对亚麻酸过氧化有一定抑制作用。
本发明与现有技术相比较,具有以下有益效果:
本发明在长寿老人的粪便中分离筛选出了青春双歧杆菌菌株ZJ2,并对该菌株的环境耐受情况、抑菌作用、自由基的清除能力和还原力大小进行试验;经过试验发现,该菌株对胃肠道内的致病菌具有一定的抑制作用,有助于恢复胃肠道微生物菌群的稳定和协调。
附图说明
图1为青春双歧杆菌ZJ2菌株菌落示意图。
图2为青春双歧杆菌ZJ2菌株与相关种的16S rDNA序列系统发育树。
图3为青春双歧杆菌ZJ2菌株对酸的耐受情况的柱状统计图。
图4为青春双歧杆菌ZJ2菌株对胆盐的耐受情况的柱状统计图。
图5为青春双歧杆菌ZJ2菌株对几种病原菌的抑菌作用效果图。
图6为青春双歧杆菌ZJ2菌株对DPPH自由基的清除能力的柱状统计图。
图7为青春双歧杆菌ZJ2菌株的还原力大小的柱状统计图。
图8为青春双歧杆菌ZJ2菌株对亚麻酸过氧化的抑制作用的柱状统计图。
图9为青春双歧杆菌ZJ2菌株对ABTS+自由基的清除能力的柱状统计图。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1分离筛选鉴定
在厌氧工作站里,取安徽省亳州市谯城区的健康百岁长寿老人的粪便样品2g,用无菌生理盐水进行梯度稀释,分别涂布于BL、BBL、BS、TPY及 MRS-L等固体培养基,于37℃厌氧培养24-48h后选取菌落数量适中的平板,挑取乳白色圆形菌落进行划线分离2-3次,得到纯化的单菌落。将分离得到的菌株分别进行液体培养,所得到的菌体用于抽取菌株的基因组DNA,利用设计的引物扩增其基因片段,将扩增的16S rDNA基因片段测序,测序结果经NCBI数据库Blast比对,初步判断菌株的种属分类。
根据《益生菌类保健食品申报与审评规定(试行)》,判定分离筛选出的菌株属于益生菌类保健品的菌株,将筛选纯化后的菌株接种于TPY肉汤中,菌液与50%的甘油(生理盐水配置)等体积混合,保存于-80℃冰箱备用。
⑴定性鉴定
按照《伯杰细菌鉴定手册》和《常见细菌系统鉴定手册》,并结合之前观察的菌株形态特征和生理生化特性,对分离得到的ZJ2菌株进行初步鉴定,可初步判断该菌株归属于双歧杆菌。
该菌株为厌氧的革兰氏阳性细菌,菌株细胞长而稍弯曲,有多种形态,如Y型、V型、长杆状、弯曲弧状。该菌株菌落光滑、凸圆、边缘完整、乳脂白色、具有柔软的质地,如图1所示。该菌株的生理生化特征如表1所示。
表1 ZJ2菌株API 20鉴定结果
符号说明:“+”,阳性;“-”,阴性。
使用细菌基因组提取试剂盒提取基因组DNA,PCR扩增目标序列后进行测序,将测序结果在数据库中进行序列比对,获得与其相似性较高的其他菌株的基因序列,详见序列表。
使用MEGA 7.0软件构建系统发育树,如图2所示,确定该菌的种属分类地位,由进化树图可看出ZJ2菌株定位在青春双岐杆菌。
采用MEGA7.0软件,邻位连接法显示“青春双歧杆菌ZJ2菌株”与相关种的16S rDNA序列系统发育树,进行1000次的相似度重复计算,图中发育树节点只显示Bootstrap值大于50%数值,上标的“T”表示模式菌株。
该菌株已于2019年11月04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,保藏单位地址:北京市朝阳区北辰西路1号院3号;保藏编号为CGMCC NO.18901,建议的分类学名称:青春双歧杆菌(Bifidobacterium adolescentis)。
实施例2动物毒性试验
选用北京维通利华实验动物技术有限公司繁殖的SPF级ICR小鼠(许可证编号SCXK(京)2016-0006)80只,雌雄各半,雄性小鼠体重范围16.0-25.0g,雄性小鼠体重范围16.0-25.0g。在北京维通利华实验动物技术有限公司动物房 (许可证编号为SYXK(京)2017-0022)饲养。
将送检的经菌种鉴定后的青春双歧杆菌ZJ2菌株转接到MRS肉汤培养基中,置于36℃,厌氧培养2天后,作为培养物原液备用。取部分培养物原液浓缩5倍,作为培养物5倍浓缩液备用。
取ICR小鼠80只,雌雄各半,分别随机为4组,每组10只。剂量组设置为:培养基空白对照组,培养基5倍浓缩空白对照组,培养物原液组,培养物5倍浓缩组。将受试物以20.0ml/kg BW剂量给小鼠灌胃,连续灌胃3天,观察7天,记录实验过程中小鼠的体重变化,中毒表现及死亡情况。
青春双歧杆菌ZJ2菌株培养物原液经梯度稀释后活菌计数结果(接种量为0.1ml),计数结果为培养物原液中含有青春双歧杆菌ZJ2菌株的数量为4.7×107 CFU/ml。
青春双歧杆菌ZJ2菌株(Bifidobacterium adolescents ZJ2)培养物原液和 5倍浓缩液与各自对照组相比,对小鼠体重增长的影响无显著性差异(p> 0.05),同时未观察到受试小鼠有毒性反应或死亡,结果如表2、表3和表4 所示。
表2青春双歧杆菌ZJ2菌株(Bifidobacterium adolescentsZJ2)培养物
对雌性小鼠体重的影响
(Mean±SD)
结果说明:雌性小鼠的初始体重和终体重在培养物组与其相应对照组间比较均无显著性差异(p>0.05),即在试验期间青春双歧杆菌ZJ2菌株(B ifidobacteriumadolescents ZJ2)培养物对雌性小鼠的体重无影响。
表3青春双歧杆菌ZJ2菌株(Bifidobacterium adolescents ZJ2)培养物
对雄性小鼠体重的影响
(Mean±SD)
结果说明:雄性小鼠的初始体重和终体重在培养物组与其相应对照组间比较均无显著性差异(p>0.05),即在试验期间青春双歧杆菌ZJ2菌株( Bifidobacteriumadolescents ZJ2)培养物对雄性小鼠的体重无影响。
表4青春双歧杆菌ZJ2菌株(Bifidobacterium adolescents ZJ2)培养物
对小鼠的急性毒力作用
结果说明:将青春双歧杆菌ZJ2菌株(Bifidobacterium adolescents ZJ2) 培养物原液和5倍浓缩液,以20.0ml/kg BW的剂量给小鼠连续灌胃3天,观察7天,未观察到受试小鼠有毒性反应或死亡,表明ZJ2菌株对小鼠无急性毒力作用。
实施例3耐受性实验
胃液的pH一般维持在2.5-3.5之间,进食后食物在胃内停留时间为2-3h。胃液有强酸性,可激活胃蛋白酶原,杀死随食物进入胃内的一些细菌,胆盐由肝脏合成储存于胆囊分泌到肠道,通常情况下肠道内的胆盐浓度大约在 0.03%-0.3%范围内。益生菌是对人体健康有着极其重要的作用,能帮助肠道抵御疾病及维持肠道正常功能,因此若想要益生菌正常发挥其生理功能则其必须具备耐受胃液的强酸和消化道的胆盐环境的特性。
实验方法:
1、将筛选出的青春双歧杆菌菌株培养在37℃的厌氧条件下,接种量为 4%,分别取不同pH的培养基培养0h、2h的菌液,稀释适当比例后涂平板培养24h计活菌数。
结果表明:如图3所示,ZJ2菌株可耐受pH 2.0的酸性环境下孵育2h仍可正常存活,耐酸性良好。
2、将筛选出的青春双歧杆菌ZJ2菌株培养在37℃的厌氧条件下,双歧杆菌的接种量为4%,分别取不同浓度胆盐的TPY肉汤,培养0h、3h的菌液稀释适当比例,涂平板培养24h计数。评价双歧杆菌的胆盐耐受性,
结果表明:由图4可看出ZJ2菌株可在0.3%的胆盐环境中仍有一定的存活率。
实施例4抑菌能力实验
实验方法:牛津杯法测定双歧杆菌对大大肠杆菌ATCC33849、沙门氏菌ATCC14028、绿脓杆菌ATCC9027、肺炎克雷伯杆菌ATCC700603、金黄色葡萄球菌NCTC8325、单增李斯特菌ATCC19115的抑制作用。
甘油保存的病源指示菌接种到TSB液体培养基中培养8h,稀释至OD600=1.0,再用生理盐水1:103稀释,吸取稀释的菌悬液200μl涂布于TSB固体培养基进行抑菌实验。牛津杯加入200μl培养后的双歧杆菌菌液以及无细胞上清液,以未接种的培养基作为对照,每组重复三次,37℃培养24h,测量抑菌圈的大小,评价双歧杆菌的抑菌能力。
结果表明:如表5所示,青春双歧杆菌ZJ2对大肠杆菌ATCC33849、沙门氏菌ATCC14028、绿脓杆菌ATCC9027、肺炎克雷伯杆菌ATCC700603、金黄色葡萄球菌NCTC8325、单增李斯特菌ATCC19115均有抑制作用,其中对单增李斯特菌的抑制能力最强,对肺炎克雷伯杆菌抑制能力最弱。
表5青春双歧杆菌ZJ2对几种病原菌的抑制作用
a抑菌圈的值为重复三次的平均值±标准误差。
实施例5抗生素敏感性实验
抗生素通过干扰细胞壁合成(青霉素、万古霉素)、损伤胞膜(氨基糖苷类:链霉素、卡那霉素)、抑制细菌蛋白质合成(四环素类:四环素;大环内酯类:红霉素)和抑制核酸合成(利福平;喹诺酮类:诺氟沙星)等方式发挥抑菌或杀菌作用,通过MIC法检测双歧杆菌对8种抗生素的敏感性。
实验方法:
受试菌液准备:受试菌株按1%接种培养18h,测定其OD600nm值,用 TPY培养基稀释菌液浓度为5.0×105-1.0×106cfu/ml。
抗生素准备:用蒸馏水或无水乙醇配置抗生素的母液,再用TPY肉汤将各抗生素稀释到实验所需最大浓度,再用TPY肉汤倍比稀释8个梯度。
96孔板每孔加75μl不同浓度的抗生素和75μl已准备好的菌液混匀,每个抗生素作3个平行,于37℃厌氧培养24h后观察菌液生长情况。菌液未长的最低浓度记作该抗素的MIC。
结果表明:如表6所示,菌株ZJ2对利万古霉素(Van)、红霉素(Ery) 以及四环素(Tet)敏感。
表6青春双歧杆菌ZJ2对8种抗生素的耐药性
注:EFSA即欧洲食品安全局,≧标准值判定为“耐受”;<标准判定为“不耐受”。
实施例6抗氧化能力实验
实验方法:
将37℃厌氧培养18h后的菌株培养液经6000r/min离心10min获得双歧杆菌菌体细胞及无细胞上清液备用。收集的菌体用pH 7.4的PBS缓冲液洗涤菌体细胞并重悬,调整其浓度为1×109CFU/ml作为完整的菌体细胞在适当的缓冲液中重悬。PBS快速洗涤菌体两次并进行冰浴超声,然后8000r/min 离心20min,收集上清液即为无细胞提取物,备用。
(1)清除DPPH自由基能力测定
取0.8ml样品中加入1ml 0.2mmol DPPH无水乙醇溶液,室温避光反应 30min,6000r/min离心10min,取上清液在波长517nm处测定吸光度。对照组以等体积蒸馏水代替样品溶液,并以等体积蒸馏水和无水乙醇混合液空白调零。
计算公式:清除率=[1-A517(样品)/A517(对照)]×100%。以0.1mg/ml 的抗坏血酸溶液为阳性对照。
结果表明:如图6所示,完整细胞和无细胞提取物对DPPH的清除效果均超过了60%。
(2)还原力的测定
取0.5ml样品加入0.5ml pH 6.6的PBS缓冲液(0.02M)和0.5ml铁氰化钾(1%),混匀后于50℃水浴中保温20min,迅速冷却,加0.5ml三氯乙酸(10%),混匀后3000r/min离心10min。移取1.5ml上清液于试管中,加入0.2ml FeCl3(0.1%),700nm测定吸光度。用PBS作为阴性对照,吸光度值越大表示待测样品的还原能力越强,制作半胱氨酸还原力的标准曲线,用等效半胱氨酸的量表示还原力大小。
结果表明:青春双歧杆菌ZJ2具有良好的还原力,如图7所示,青春双歧杆菌ZJ2还原力大小的范围为69.57-56.83(μM)。
(3)抑制亚麻酸的过氧化作用(TAALA)
1.5%(v/v)亚麻酸(96%的乙醇溶解)用生理盐水1:125v/v稀释,吸取 400ml稀释后的溶液加入15ml 0.01%十二烷基硫酸钠(w/v)和45ml的样品溶液混匀。反应开始通过添加100ml FeSO4·7H2O(0.1g/L),混匀后于37℃反应60min后,加入35ml 0.25%(v/v)丁基羟基甲苯溶液终止反应。然后,加入500ml 0.1mol/L的醋酸缓冲液(pH3.5)和1ml的0.6%(w/v)硫代巴比妥酸(TBA)溶液,80℃加热40min,混合液冷却至室温,加入0.5ml预冷的Hcl(5mol/L)和2ml预冷的正丁醇。3000r/min离心10min测定了534nm处上清液的吸光度。以生理盐水代替样品作为空白对照。TAALA表示为抑制亚麻酸过氧化的百分率(TAALA),定义如下:
抑制率=[1-A534(样品)/A534(对照)]。
结果表明:如图8所示,青春双歧杆菌ZJ2对抑制亚麻酸的过氧化具有良好的作用,其中,完整细胞的抗氧化能力要优于无细胞上清液。
(4)清除ABTS+自由基能力测定
7.4mmol/L ABTS和2.6mM过硫酸钾混合,混合物在黑暗的室温下放置 12-16小时,直到达到稳定的氧化状态(室温保存2d)。PBS缓冲液(pH7.4) 或95%乙醇稀释40~50倍,在734nm吸光度下OD值达到0.7±0.02,备用。 0.2ML工作液与10ml样品混合置于冰上,6min后测OD。抑制率通过Trolox 的浓度表示。
结果表明:如图9所示,ZJ2菌株完整细胞和无细胞提取物的等效Trolox 量分别为359.87±21.17和430.00±9.15(μM)。
实例7胆固醇降解实验
实验方法:
MRS-CHOL(胆固醇MRS培养基):MRS-L培养基(MRS专用培养基 +0.5g/L半胱氨酸)+0.1mg/ml胆固醇,过滤除菌。胆固醇标准液:即胆固醇浓度为1.0mg/ml,100mg胆固醇,用少量无水乙醇溶解并定容至100ml,即胆固醇标准溶液,放置0℃~4℃密封。硫酸铁铵显色剂:4.463g硫酸铁铵溶于100ml 85%磷酸中,即为储备液,再取10ml该储备液用浓硫酸定容至100ml,即为显色剂。
取培养18h后的双歧杆菌,按1%的接种量接种至MRS-CHOL培养基中,等体积未接种的MRS-CHOL培养基作为空白对照,37℃厌氧培养48h,测定双歧杆菌对胆固醇的降解率:取3ml菌液6000r/min离心10min,取上清液2ml 于50ml离心管中,加无水乙醇至10ml,充分混匀,4500r/min离心10min,取上清液1ml至12ml离心管中,缓缓加入2ml硫酸铁铵显色剂(若直接用菌液上清液加显色剂显示颜色很深),立即摇匀,室温下放置30min后测各管的OD560nm值,计算双歧杆菌对胆固醇的降解率。
W=(A-B)/A×100%
注:A:未接种双歧杆菌的MRS-CHOL培养基中胆固醇的含量(即空白对照),mg;B:双歧杆菌在MRS-CHOL培养基中发酵后胆固醇的含量,mg;W: 胆固醇降低率,%。
结果表明:如表7所示,外源加入的0.1mg/ml胆固醇孵育48h后青春双歧杆菌ZJ2的胆固醇降解率为29.15%,表明青春双歧杆菌ZJ2具有良好的降解胆固醇的能力。
表7降解胆固醇能力的测定
对于本领域技术人员而言,显然本发明不限于上述示范性实施例的细节,而且在不背离本发明的精神或基本特征的情况下,能够以其他的具体形式实现本发明。因此,无论从哪一点来看,均应将实施例看作是示范性的,而且是非限制性的,本发明的范围由所附权利要求而不是上述说明限定,因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。不应将权利要求中的任何附图标记视为限制所涉及的权利要求。
SEQUENCE LISTING
<110> 卓源健康科技有限公司
<120> 一种青春双歧杆菌菌株及其应用
<130> 2019.10.22
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1428
<212> DNA
<213> 1
<400> 1
tgcttacaca tgcaagtcga acgggatccc aggagcttgc tcctgggtga gagtggcgaa 60
cgggtgagta atgcgtgacc gacctgcccc atacaccgga atagctcctg gaaacgggtg 120
gtaatgccgg atgctccagt tgaccgcatg gtcctctggg aaagcttttg cggtatggga 180
tggggtcgcg tcctatcagc ttgatggcgg ggtaacggcc caccatggct tcgacgggta 240
gccggcctga gagggcgacc ggccacattg ggactgagat acggcccaga ctcctacggg 300
aggcagcagt ggggaatatt gcacaatggg cgcaagcctg atgcagcgac gccgcgtgcg 360
ggatgacggc cttcgggttg taaaccgctt ttgactggga gcaagccctt cggggtgagt 420
gtacctttcg aataagcacc ggctaactac gtgccagcag ccgcggtaat acgtagggtg 480
caagcgttat ccggaattat tgggcgtaaa gggctcgtag gcggttcgtc gcgtccggtg 540
tgaaagtcca tcgcttaacg gtggatccgc gccgggtacg ggcgggcttg agtgcggtag 600
gggagactgg aattcccggt gtaacggtgg aatgtgtaga tatcgggaag aacaccaatg 660
gcgaaggcag gtctctgggc cgtcactgac gctgaggagc gaaagcgtgg ggagcgaaca 720
ggattagata ccctggtagt ccacgccgta aacggtggat gctggatgtg gggaccattc 780
cacggtctcc gtgtcggagc caacgcgtta agcatcccgc ctggggagta cggccgcaag 840
gctaaaactc aaagaaattg acgggggccc gcacaagcgg cggagcatgc ggattaattc 900
gatgcaacgc gaagaacctt acctgggctt gacatgttcc cgacagccgt agagatacgg 960
tctcccttcg gggcgggttc acaggtggtg catggtcgtc gtcagctcgt gtcgtgagat 1020
gttgggttaa gtcccgcaac gagcgcaacc ctcgccctgt gttgccagca cgtcatggtg 1080
ggaactcacg ggggaccgcc ggggtcaact cggaggaagg tggggatgac gtcagatcat 1140
catgcccctt acgtccaggg cttcacgcat gctacaatgg ccggtacaac gggatgcgac 1200
accgcgaggt ggagcggatc ccttaaaacc ggtctcagtt cggattggag tctgcaaccc 1260
gactccatga aggcggagtc gctagtaatc gcggatcagc aacgccgcgg tgaatgcgtt 1320
cccgggcctt gtacacaccg cccgtcaagt catgaaagtg ggtagcaccc gaagccggtg 1380
gcccaacctt ttggggggag ccgtctaagg gagaccttcc gaggttta 1428
Claims (6)
1.一种青春双歧杆菌菌株,其特征在于:所述菌株由健康百岁长寿老人的粪便中分离筛选得到的,命名为青春双歧杆菌(Bifidobacterium adolescents)ZJ2,且已于2019年11月04日保藏于中国微生物菌种保藏中心,保藏编号为CGMCC NO.18901。
2.一种菌剂,其特征在于:所述菌剂的活性成分中含有权利要求1所述青春双歧杆菌菌株ZJ2。
3.一种组合物,其特征在于:所述组合物的成分中含有权利要求2所述青春双歧杆菌菌株ZJ2的菌剂。
4.一种权利要求3所述的组合物在食品中的应用。
5.一种权利要求3所述的组合物在保健产品中的应用。
6.一种权利要求3所述的组合物在制备抑制胃肠道致病菌药物中的应用。
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