CN115109734A - 一株具有缓解高尿酸功能敏捷乳杆菌b13t4及其应用 - Google Patents
一株具有缓解高尿酸功能敏捷乳杆菌b13t4及其应用 Download PDFInfo
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- CN115109734A CN115109734A CN202211050279.5A CN202211050279A CN115109734A CN 115109734 A CN115109734 A CN 115109734A CN 202211050279 A CN202211050279 A CN 202211050279A CN 115109734 A CN115109734 A CN 115109734A
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- lactobacillus agilis
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- lactobacillus
- agilis
- hyperuricemia
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Abstract
本发明提供了一株具有缓解高尿酸功能的敏捷乳杆菌(Lactobacillus agilis)B13T4及其应用,涉及微生物及其应用技术领域。所述敏捷乳杆菌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.24786。本发明提供的敏捷乳杆菌菌株B13T4是从成年人粪便中分离和筛选得到,能够显著降低高尿酸症模型小鼠的嘌呤代谢关键酶黄嘌呤氧化酶水平及尿酸含量,并改善高尿酸血症引起的炎症因子水平升高,对治疗或预防改善高尿酸血症具有较好的开发利用前景。
Description
技术领域
本发明涉及微生物及其应用技术领域,尤其是涉及一株具有缓解高尿酸功能敏捷乳杆菌(Lactobacillus agilis)B13T4及其应用。
背景技术
高尿酸血症是嘌呤代谢紊乱引起的代谢异常综合征,当血尿酸超过其在血液或组织液中的饱和度时可形成尿酸钠结晶,该结晶沉积在关节可导致痛风,如沉积在肾脏,则引起尿酸性肾病。高尿酸血症和痛风是慢性肾病、心脑血管疾病和糖尿病等的独立危险因素。随着人们生活水平的不断提高、饮食结构和生活习惯的改变,高尿酸血症与痛风的发病率逐年上升,发病年龄呈现低龄化。高尿酸血症及痛风是继糖尿病之后又一常见代谢性疾病,人群基数极大。高尿酸血症对疾病管理要求较高,是一个连续的过程,需要长期、甚至是终生进行监测与管理。在这一过程中,将血尿酸控制在理想范围是降低痛风及其相关并发症发生的根本。
血尿酸的控制,一方面需要在饮食上限制高嘌呤、高果糖饮食,另一方面可以通过药物治疗。尽管现有药物能够较好的降低尿酸,但仍有一些潜在风险,例如别嘌醇在亚洲人群的超敏反应发生率高、非布司他在心血管疾病患者中使用时需考虑风险性以及苯溴马隆对尿液PH值监测的要求较高。因此,寻找更加安全有效的将尿酸药物尤为重要。
目前,已有研究发现栖居于人类胃肠道系统内的肠道微生物与高尿酸血症及痛风的发生发展密切相关。这提示了肠道微生物中的一些物种或物质能够帮助降低血尿酸。因此,通过合适的高通量筛选体系,对于肠道微生物菌库进行功能挖掘是当期该领域的研究热点,更具体地是寻找具有高效调节代谢稳态功能的菌株。由于肠道菌群尤其是益生菌安全性较高,分离纯化成本低,如果能够明确其降尿酸功能,并确定其降尿酸机制,那么作为下一代改善高尿酸产品(如食品或/和药物)的市场前景将十分广阔。
鉴于此,特提出本发明。
发明内容
本发明的目的在于提供一株敏捷乳杆菌(Lactobacillus agilis)B13T4及其应用,本发明的菌株能够降低嘌呤代谢关键酶黄嘌呤氧化酶水平及尿酸含量,并改善高尿酸血症引起的炎症因子水平升高。
本发明提供的技术方案如下:
在一个方面,本发明提供了一株敏捷乳杆菌(Lactobacillus agilis)菌株B13T4,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.24786。
本发明从长期生活在中国海南澄迈的百岁老人的粪便中分离并筛选到一株敏捷乳杆菌B13T4,通过黄嘌呤氧化酶活性体外实验证明,本发明的菌株(敏捷乳杆菌上清液)对黄嘌呤氧化酶具有较高的抑制活性,表现出治疗高尿酸症的潜能。将本发明将菌株施用于小鼠,观察菌株降尿酸的功效发现,喂养本发明的敏捷乳杆菌B13T4的小鼠的XOD的活性明显降低,其效果优于阳性药别嘌醇,避免了尿酸过量产生;并且高尿酸模型组小鼠的炎症因子的水平明显降低,敏捷乳杆菌B13T4能够缓解尿酸代谢异常引起的炎症反应。
在一个方面,本发明提供了一种菌剂,所述菌剂含有前述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4。
在具体的实施方案中,所述菌剂为固体制剂或液体制剂;所述固体菌剂为采用冷冻干燥法制备而成的粉剂。
在具体的实施方案中,所述菌剂中,敏捷乳杆菌B13T4的含量≥1×106CFU/mL或≥1×106CFU/g。
在一个方面,本发明提供了所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或所述菌剂在抑制黄嘌呤氧化酶中的应用。进一步提供了所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或所述菌剂在改善嘌呤代谢方面或制备相关产品中的应用。
在一个方面,本发明提供了所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或所述的菌剂在制备降尿酸的产品中的应用。本发明的菌株和菌剂可用于治疗和预防高尿酸血症,例如制备预防或缓解高嘌呤膳食即肌苷引起的高尿酸血症的药物,或制备用于预防或缓解高尿酸血症引起的肠道菌群紊乱的药物。
在一个实施方案中,所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或所述的菌剂用于制备降低高尿酸血症引起的炎症的药物。
在一个方面,本发明提供了所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或所述的菌剂用于治疗或预防高尿酸血症和/或痛风的产品中的应用或在制备痛风疾病辅助治疗药物中的应用。
鉴于本发明菌株的对黄嘌呤氧化酶的抑制活性以及降尿酸活性,本发明的菌株及其衍生产品能够用于制备具有降尿酸功能或具有抗痛风功能产品。并且进一步地可以单独使用或与其他具有降尿酸功能或具有抗痛风功能的产品联合应用。这些产品可制备为微生物制剂、功能性食品、保健品或药物。所述其他具有降尿酸功能或具有抗痛风功能的产品为别嘌呤醇、非布司他、苯溴马隆和丙磺舒中的一种或多种。
在一个实施方案中,所述敏捷乳杆菌为灭活的或未灭活的发酵上清液、菌悬液和细胞破碎物上清液中的一种或多种。
在一个实施方案中,所述灭活的方式为巴氏杀菌。
在另一个方面,本发明提供了一种药物,所述药物含有所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4和药学上可接受的载体。所述药学上可接受的载体包括但不限于:填充剂、润湿剂、崩解剂、粘合剂或润滑剂中的一种或多种。
在一个实施方案中,所述填充剂为微晶纤维素、乳糖、甘露醇、淀粉或糊精中的一种或多种;所述润湿剂为乙醇或甘油中的一种或多种;所述崩解剂为羧甲基淀粉钠、交联羧甲基淀粉钠、交联聚维酮或低取代羟丙基纤维素中的一种或多种;所述粘合剂为淀粉糊、糖浆、饴糖或液状葡萄糖中的一种或多种;所述润滑剂为硬脂酸镁、滑石粉或二氧化硅中的一种或多种。
在一个实施方案中,所述药物为供口服的剂型;优选地,所述药物的剂型为溶液剂、悬浮液剂、乳剂、粉末剂、锭剂、丸剂、糖浆剂、口含锭剂、片剂以及胶囊剂。
在另一个方面,本发明提供了一种改善嘌呤代谢的方法,所述方法是将前述所述敏捷乳杆菌(Lactobacillus agilis)B13T4菌株加入到含有嘌呤的体系中。所述方法为非疾病的诊断或治疗目的。
本发明的敏捷乳杆菌(Lactobacillus agilis)B13T4的培养方法为在MRS肉汤培养基中于35-37℃条件下培养。所述的培养的时间为18-24小时。
生物样品保藏信息:敏捷乳杆菌(Lactobacillus agilis)B13T4,该菌株已于2022年4月27日保藏于中国微生物菌种保藏管理委员会普通微生物菌种保藏中心,保藏号为:CGMCC No.24786;保藏地址:中国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。经保藏中心于2022年4月27日检测为存活菌株。
有益效果:
1.本发明提供的敏捷乳杆菌能够明显抑制黄嘌呤氧化酶活性,并且可以显著降低高尿酸症模型小鼠的嘌呤代谢关键酶黄嘌呤氧化酶水平及尿酸含量,并改善高尿酸血症引起的炎症因子的升高,缓解高尿酸血症。
2. 本发明的敏捷乳杆菌可高效缓解或治疗高尿酸血症或痛风的病症,可用于制备防治高尿酸血症或/和痛风、降解核苷、抑制黄嘌呤氧化酶的功能性食品或药物,应用前景非常广阔。敏捷乳杆菌及含敏捷乳杆菌的发酵制品在制备防治高尿酸血症(降尿酸)、抗痛风、抗炎症、改善肠道菌群多样性方面有广泛的潜力。
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为本发明提供的敏捷乳杆菌B13T4的质谱鉴定图;
图2为本发明中实施例提供的敏捷乳杆菌B13T4的细菌培养物上清液对黄嘌呤氧化酶的体外抑制作用,别嘌醇作为阳性药,n=3;
图3为示出了灌胃本发明中敏捷乳杆菌B13T4后降低高尿酸小鼠的黄嘌呤氧化酶活性,别嘌醇作为阳性药,n=8,#,p<0.05; ##,p<0.01; ###,p<0.001,模型组vs正常对照组;*,p<0.05;**,p<0.01;***,p<0.001,B13T4,别嘌醇组vs正常对照组;
图4为示出了灌胃本发明中敏捷乳杆菌B13T4后对高尿酸小鼠血清尿酸UA水平的影响,别嘌醇作为阳性药,n=8,#,p<0.05; ##,p<0.01;###,p<0.001,模型组vs正常对照组;*,p<0.05;**,p<0.01;***,p<0.001,B13T4,别嘌醇组vs正常对照组;
图5为示出了灌胃本发明中敏捷乳杆菌B13T4后抑制高尿酸小鼠的炎症因子IL-1β、IL-6、TNF-α水平,别嘌醇作为阳性药,n=8,#,p<0.05;##,p<0.01;###,p<0.001,模型组vs正常对照组;*,p<0.05;**,p<0.01;***,p<0.001,B13T4, 别嘌醇组vs正常对照组。
具体实施方式
下面将结合实施例对本发明的技术方案进行清楚、完整地描述,显然,所描述的实施例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. 敏捷乳杆菌B13T4的分离和培养
1.1敏捷乳杆菌B13T4分离鉴定
采集长期生活在中国海南澄迈世界长寿之乡的百岁老人的粪便作为筛选样品,采集对象人群在采集前未服用过抗生素类药物,无益生菌服用史,无胃肠病史。
将采集来的粪便样品稀释液5 mL,10倍系列稀释至10-6,每个稀释浓度取200 μL涂布于选择性MRS培养基(蛋白胨 10 g,牛肉膏 10 g,酵母膏 5 g,柠檬酸氢二铵 2 g,葡萄糖20 g,吐温80 1 mL,乙酸钠5 g,磷酸氢二钾 2 g,硫酸镁 0.58 g,硫酸锰0.25 g,琼脂18 g,蒸馏水1000 mL),37℃厌氧培养24-48 h,分离获得平板上不同的单菌落,然后将每个单菌落分别在新的MRS固体培养基平板上进行划线操作,37℃厌氧培养48 h后得到纯化菌落,将每个纯化单菌落涂布在质谱板上,分别加入裂解液和基质干燥后在MALDI-TOF MS1000质谱仪(Autobio,郑州安图生物科技有限公司)上进行质谱上机鉴定,鉴定完成后将各菌株于-80℃冻存在本公司的菌种资源库中,以待后用。
其中本发明所涉及的敏捷乳杆菌B13T4菌株鉴定结果如图1所示,B13T4菌株与敏捷乳杆菌种具有极高的相似性,据此将其命名为敏捷乳杆菌B13T4菌株。
敏捷乳杆菌B13T4菌液的制备
(1)取-80℃冻存菌液涂布于MRS固体平板在37℃条件下倒置培养24-48h后,取单菌落接种于MRS液体培养基中,37℃培养18-24h,得到一代菌液;取一代菌液10 %(v/v)接种至新鲜MRS液体培养基,37℃培养18-24h得到二代菌液;取二代菌液10 %(v/v)接种于新鲜的MRS液体培养基中,37℃培养18-24h得到工作菌液。
(2)将敏捷乳杆菌B13T4的工作菌液置于13000 rpm,4℃的条件下离心15 min后,收集上清,即为菌株发酵上清液样品,可用于以下实施例中的体外酶活实验;将敏捷乳杆菌B13T4的工作菌液置于13000 rpm,4℃的条件下离心15 min后,弃去上清,收集沉淀,用生理盐水重悬,获得具有活菌体的活菌液,浓缩计数达1×109 CFU/mL以上即可用于后续实施例中的动物实验。
活菌液还可以通过本技术领域中的其他方式获得,只要能从培养液中富集菌体即可。例如可以通过离心和/或过滤的方法实现。
实施例2、黄嘌呤氧化酶活性体外实验
本发明使用的黄嘌呤Xan、黄嘌呤氧化酶((xanthine oxidase,XOD)、别嘌醇Allopurinol均购自索莱宝,使用方法均按照说明书的方法,具体使用方法如下:
黄嘌呤溶液配制:
6 mM储液:精密称取黄嘌呤7.3 mg,在烧杯中提前加入0.6 mL的1 mol/L NaOH和3ml去离子水,将黄嘌呤粉末加入到烧杯中,轻轻摇晃至黄嘌呤完全溶解并混匀溶液,使用PH测量仪测量溶液pH值,缓慢滴入1 mmol/L HCl调整pH值至7.5-8.5;将调整完PH值的黄嘌呤溶液倒入洗净的量筒中,加入去离子水至终体积8 mL。使用时加入反应体系中的溶液至浓度为0.6 mmol/L,每天新鲜配制。
黄嘌呤氧化酶溶液配制:根据瓶身提供的信息,计算原始酶浓度。加入体系工作液浓度为0.074 U/mL左右,需将原始酶液用PBS稀释。
别嘌醇溶液配制:精密称取0.034 g别嘌醇,根据试剂溶解度计算可得,使用最小体积0.5 mL 1M NaOH作为溶剂,加DMSO 6mL,加PBS配制成10 mmol/L贮备液25 mL,调PH至8.5左右,于4℃避光保存。使用时,用PBS稀释至0.25mmol/L。
实验过程:黄嘌呤在黄嘌呤氧化酶催化下能够生成尿酸。本发明在该反应中加入将本发明提供的敏捷乳杆菌上清液,观察本发明提供的菌株对黄嘌呤氧化酶的影响,具体样品设置包括:敏捷乳杆菌B13T4组及不加XOD的空白对照组,阳性药组(别嘌醇组)及不加XOD的空白对照组,阴性对照组(培养基MRS)及不加XOD的空白对照组,每组设置3个复孔。
实验操作如下:在96孔板中,实验组分别加入以下溶液,溶液加入的顺序为15 μL敏捷乳杆菌上清液或敏捷乳杆菌培养基MRS→PBS→10 μL黄嘌呤氧化酶→50 μL黄嘌呤,其中所有空白对照组不加黄嘌呤氧化酶,所有组别补充PBS至200 μL。加样结束后,温箱37℃孵育30 min。使用酶标仪在290 nm处测吸光度值,该数值用以计算敏捷乳杆菌抑制黄嘌呤氧化酶的效力。
根据以上分组设置,分别计算每组的∆A:
B13T4组:∆A (B13T4) =A290 (B13T4)–A290 (Ctrl)
别嘌醇组:∆A (别嘌醇) =A290 (别嘌醇)–A290 (Ctrl)
阴性对照组:∆A (阴性对照组)=A290 (MRS)– A290 (Ctrl)
XOD黄嘌呤氧化酶抑制率 (%) =[1-∆A (B13T4/别嘌醇)/∆A(阴性对照组)]*100%
根据图2所示,阳性药别嘌醇对黄嘌呤氧化酶的平均抑制率为90.140%,敏捷乳杆菌B13T4对黄嘌呤氧化酶的平均抑制率为72.501%,由此可见,敏捷乳杆菌B13T4发酵上清液对黄嘌呤氧化酶具有较高的抑制活性,表现出治疗高尿酸症的潜能。
实施例3、小鼠试验
本发明将菌株(活菌液)施用于小鼠,观察菌株降尿酸的功效,具体操作如下:
动物实验所用小鼠为8周龄雄性昆明小鼠36只,随机分4组,分别为正常组,高尿酸模型组,高尿酸+敏捷乳杆菌B13T4活菌液组,高尿酸+阳性药别嘌醇组。
每组8只动物,适应性饲养7天后进行建模及给药,建模试剂:奥博星产酵母浸膏,剂量20 g/kg/d。诱导建模同时给药,其中高尿酸+敏捷乳杆菌B13T4活菌液组每天给予1×109 CFU相应的B13T4活菌;高尿酸+阳性药别嘌醇组每天给予40 mg/kg的别嘌醇溶液。连续给菌4周。
4周后,用水合氯醛麻醉小鼠,眼眶取血,4℃离心后取上清作为血清样品,用于测量黄嘌呤氧化酶水平,尿酸UA含量,以及炎症因子IL-1β、IL-6、TNF-α的水平,同时处死动物,收取肝脏、肾脏等组织,-80℃低温保存。
根据图3可以看出,高尿酸模型组小鼠血液中嘌呤代谢关键酶--黄嘌呤氧化酶(XOD)含量显著升高,显示出酵母浸膏能够明显的促进高尿酸的发生,喂养本发明的敏捷乳杆菌B13T4的小鼠,其XOD的含量明显降低,其效果优于阳性药别嘌醇,因此敏捷乳杆菌B13T4能够防止自由基的形成,避免尿酸过量产生。进一步地,图4结果显示:与正常对照组比较,高尿酸模型组小鼠血液中血尿酸(UA)含量显著升高,喂养敏捷乳杆菌B13T4能够降低UA含量,其效果与阳性药别嘌醇相当。该结果表明敏捷乳杆菌B13T4能够有效降低高尿酸小鼠的尿酸水平,改善高血尿酸症。另外,如图5所示,高尿酸模型组小鼠血液中炎症因子IL-1β、IL-6、TNF-α水平显著上升,而喂养敏捷乳杆菌B13T4后,这些炎症因子的水平明显降低,其抗炎能力优于阳性药别嘌醇,表明敏捷乳杆菌B13T4能够缓解尿酸代谢异常引起的炎症反应。
根据上述实验结果,可以确定本发明提供的敏捷乳杆菌B13T4菌株具有优良的改善高尿酸功效,在制备降高尿酸、改善嘌呤代谢和预防痛风发生的产品方面具有广阔的应用前景。
最后应说明的是:以上各实施例仅用以说明本发明的技术方案,而非对其限制;尽管参照前述各实施例对本发明进行了详细的说明,本领域的普通技术人员应当理解:其依然可以对前述各实施例所记载的技术方案进行修改,或者对其中部分或者全部技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (10)
1.一株具有缓解高尿酸功能的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4,其特征在于,所述敏捷乳杆菌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.24786。
2.一种菌剂,其特征在于,所述菌剂含有权利要求1所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4。
3.权利要求1所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或权利要求2所述的菌剂在抑制黄嘌呤氧化酶中的应用。
4.权利要求1所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或权利要求2所述的菌剂在制备降尿酸的产品中的应用。
5.权利要求1所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4或权利要求2所述的菌剂在制备用于治疗或预防高尿酸血症和/或痛风的产品中的应用。
6.根据权利要求3-5中任一项所述的应用,其特征在于,所述敏捷乳杆菌为灭活的或未灭活的发酵上清液、菌悬液和细胞破碎物上清液中的一种或多种。
7.根据权利要求6所述的应用,其特征在于,所述灭活的方式为巴氏杀菌。
8.一种药物,其特征在于,所述药物含有权利要求1所述的敏捷乳杆菌(Lactobacillus agilis)菌株B13T4和药学上可接受的载体。
9.根据权利要求8所述的药物,其特征在于,所述药物为供口服的剂型;优选地,所述药物的剂型为溶液剂、悬浮液剂、乳剂、粉末剂、锭剂、丸剂、糖浆剂、口含锭剂、片剂以及胶囊剂。
10.一种改善嘌呤代谢的方法,其特征在于,所述方法是将权利要求1所述敏捷乳杆菌(Lactobacillus agilis)B13T4菌株加入到含有嘌呤的体系中。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116445372A (zh) * | 2023-06-14 | 2023-07-18 | 中国农业科学院北京畜牧兽医研究所 | 提高反刍动物瘤胃氮素转化效率的敏捷乳杆菌及其应用 |
CN116694530A (zh) * | 2023-06-28 | 2023-09-05 | 北京量化健康科技有限公司 | 一种乳杆菌组合物及其应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20160051602A1 (en) * | 2014-08-22 | 2016-02-25 | Food Industry Research And Development Institute | Novel strain of lactobacillus rhamnosus and its metabolites for use in inhibiting xanthine oxidase and treating gout |
CN113999805A (zh) * | 2021-12-06 | 2022-02-01 | 四川高福记生物科技有限公司 | 一种防治高尿酸血症的发酵乳杆菌及其组合物与应用 |
-
2022
- 2022-08-30 CN CN202211050279.5A patent/CN115109734B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20160051602A1 (en) * | 2014-08-22 | 2016-02-25 | Food Industry Research And Development Institute | Novel strain of lactobacillus rhamnosus and its metabolites for use in inhibiting xanthine oxidase and treating gout |
CN113999805A (zh) * | 2021-12-06 | 2022-02-01 | 四川高福记生物科技有限公司 | 一种防治高尿酸血症的发酵乳杆菌及其组合物与应用 |
Non-Patent Citations (6)
Title |
---|
ALLAH NAWAZKHAN 等: "Antagonistic, Anti-oxidant, Anti-inflammatory and Anti-diabetic Probiotic Potential of Lactobacillus agilis Isolated From the Rhizosphere of the Medicinal Plants", 《SAUDI JOURNAL OF BIOLOGICAL SCIENCES》 * |
CAIXIN NI等: "Lactic acid bacteria strains relieve hyperuricaemia by suppressing xanthine oxidase activity via a short-chain fatty acid-dependent mechanism", 《FOOD & FUNCTION》 * |
MARTADEC 等: "Identification of Lactobacillus strains of goose origin using MALDI-TOF mass spectrometry and 16S–23S rDNA intergenic spacer PCR analysis", 《RESEARCH IN MICROBIOLOGY》 * |
刘丹丹: "酸菜和蜜蜂胃肠道中15个乳酸菌新种的鉴定", 《中国优秀硕士学位论文全文数据库 (基础科学辑)》 * |
金方等: "降血尿酸益生菌株的筛选和降血尿酸机理的探索", 《微生物学通报》 * |
龙广云: "16个乳杆菌新种的多相鉴定", 《中国优秀硕士学位论文全文数据库 (基础科学辑)》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116445372A (zh) * | 2023-06-14 | 2023-07-18 | 中国农业科学院北京畜牧兽医研究所 | 提高反刍动物瘤胃氮素转化效率的敏捷乳杆菌及其应用 |
CN116445372B (zh) * | 2023-06-14 | 2023-09-01 | 中国农业科学院北京畜牧兽医研究所 | 提高反刍动物瘤胃氮素转化效率的敏捷乳杆菌及其应用 |
CN116694530A (zh) * | 2023-06-28 | 2023-09-05 | 北京量化健康科技有限公司 | 一种乳杆菌组合物及其应用 |
CN116694530B (zh) * | 2023-06-28 | 2023-11-07 | 北京量化健康科技有限公司 | 一种乳杆菌组合物及其应用 |
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