CN112251380A - 一株促进机体健康长寿的植物乳杆菌flpl05 - Google Patents
一株促进机体健康长寿的植物乳杆菌flpl05 Download PDFInfo
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Abstract
本发明公开了一株促进机体健康长寿的植物乳杆菌FLPL05，该菌株为植物乳杆菌FLPL05(Lactobacillus plantarum FLPL05)，于2017年9月28日保藏在中国典型培养物保藏中心(中国武汉大学)，保藏编号：CCTCC NO:M 2017763，分离自健康长寿老人粪便，为革兰氏阳性菌，生长能力强，能促进机体健康长寿，同时维持老年后肠道屏障的完整性，可降低老年后的炎症水平，可开发作为微生态菌剂，可用做食品添加剂，改善老年人胃肠道健康。
Description
技术领域
本发明属于微生物领域，涉及新的植物乳杆菌(Lactobacillus plantarum)菌株及其益生功能，尤其是用于维持肠屏障完整性和延缓衰老的功能。
背景技术
肠道菌群作为人体一大“器官”，影响着人体的发育生长、营养与健康，甚至其与代谢性疾病(肠易激综合症，炎症性肠病，心血管疾病，糖尿病，肥胖等)密切相关，肠道菌群与人体健康乃至长寿的关联性。
湖南省浏阳市高田村地理位置相对封闭，其村民不仅长寿，而且普遍没有慢性疾病。与此形成鲜明对比的是，周边地区既无长寿现象，而且心血管疾病，慢性肠胃炎和癌症等疾病时有发生。
乳酸菌可调节机体胃肠道菌群平衡，提高机体免疫力降低血清胆固醇，降血压，抗氧化抑制肿瘤发生等方面的特殊生理活性。长双歧杆菌BBMN68分离自广西巴马长寿老人粪便，其活菌液对便秘模型小鼠有润肠通便作用，提高免疫力。唾液乳杆菌FDB86可以缓解二甲肼对肠道菌群的不良影响，使肠道菌群趋近于正常状态，对结肠癌大鼠肠道菌群变化的调节作用。另外衰老会在体内累积炎症因子，容易有慢性炎症和慢性病，因此，提高健康寿命，不仅是延长寿命，同时要兼具改善慢性炎症和慢性病的功能。
目前为止，没有报道任何植物乳杆菌物种的菌株具有延长寿命同时改善炎症水平维持肠道健康。
发明内容
现在，本发明人分离鉴定了一种植物乳杆菌FLPL05菌株，是一类厌氧，不产孢子的革兰氏阳性菌，菌体呈短杆状，在MRS琼脂平板培养基中呈乳白色透明，圆形，光滑的菌落。植物乳杆菌FLPL05可以在MRS肉汤培养基上生长，是革兰氏阳性菌。
其能显著性的延长小鼠30％的寿命，维持肠道屏障的完整性，降低由衰老导致的炎症，是一株性能优良的菌种。因此，其具有很高的研究价值和应用价值。
本发明植物乳杆菌FLPL05(Lactobacillus plantarum FLPL05)，于2017 年12月7日保藏在中国典型培养物保藏中心(中国武汉大学)，编号：CCTCC NO:M 2017763。
一种微生物菌剂，含有所述植物乳杆菌菌株FLPL05，用于促进健康长寿，保护肠道屏障，降低炎症水平，预防或治疗肠屏障损伤。
包含所述的植物乳杆菌的乳制品、蔬菜制品、饮料制品和其他发酵类制品。
一种组合物，包括所述的植物乳杆菌菌株，或通过所述的方法所获得的微生物菌剂，或通过所述的方法获得的乳制品、蔬菜制品、饮料制品和其他发酵类制品。
所述的组合物是食品、保健品、药品、菌剂及饲料添加剂。
本发明所述植物乳杆菌可用于制作治疗和预防肠道疾病的组合物，促进健康长寿。
附图说明
图1植物乳杆菌FLPL05对衰老小鼠寿命的影响；
图2植物乳杆菌FLPL05干预后小鼠血清中Dextran-FITC的含量；
图3小鼠肠道H&E染色结果(A年轻小鼠，B老年小鼠，C老年小鼠灌胃 FLPL05组)；
图4植物乳杆菌FLPL05干预后血清炎症水平。
具体实施方式
为了进一步理解本发明，下面结合实施例对本发明提供的一株促进机体健康长寿的植物乳杆菌FLPL05进行说明，本发明的保护范围不受以下实施例的限制。
实施例1：植物乳杆菌FLPL05的分离与鉴定
植物乳杆菌FLPL05是从健康长寿老人的粪便中分离。取1g粪便溶于10mL 10％蛋白胨溶液中，充分溶解，滤纸滤去残渣，滤液用5000rmp离心10min，弃去上清，收集沉淀，用1mL 10％蛋白胨水重悬。按1％接种到MRS液体培养基中，过夜培养。培养液采用10倍梯度稀释，梯度稀释为10-1、10-2、10-3、 10-4、10-5、10-6、10-7、10-8和10-9，每个梯度取1mL分别均匀涂布于MRS肉汤的琼脂平板上，37℃厌氧培养48h。挑取单菌落，在平板上划线分离，反复进行纯化培养，用革兰氏染色法观察菌落形态选出革兰氏阳性菌。将分离得到的菌株液体培养后，平板培养，用接种环挑取单一的菌落作为模板，进行菌落PCR。将阳性PCR产物送至公司测序。
将所分离菌菌株的16S rRNA碱基序列分析测序，结果：
ACTTGACGTTAATACTGACGCTATTACCTTAACTGGTAGTGATGCTGATATTTCTATTGAAA CCACCATTCCAGCAAGTGATGACAACAATACGTTAGTCGTTGAAGATGCCGGTTCAATCGTTTTAC CTGCCCGTTTCTTTAGTGAAATCGTAAAAAAATTACCAGAAGACACCATGACTGTTAACGTGGTCG ACGGTTTTCAAACACAAATTACTTCAGGGGCCGCTTCCTTTACAATCAACGGCTTAGACCCTGAAA ACTACCCACACTTACCTGAAATTGATACGACTAATACCATCACGTTAGCTGGGGATGTACTCAAAG AGTTGATTGGTCAGACCGTTATTGCCGTTTCCAATCAGGAAAGTCGACCAATTCTGACCGGGGTTC ATTTTATCTTAGCTAACGGTGAGTTCTTAGCGGTCGCAACCGATTCACACCGCCTAAGTCAACGCC GGATCAAGTTGCCAGAAGCCAATAATGCCAATTATGATGTAATCATTCCTGGTAAAAGTTTGACAG AATTATCTCGGATGATCGGCGACAATAATCCTGACGTGCAGATGCGGTTATCAGAAAACCAAGTCC TATTTGTGCTGGGGAATACGTCCTTCTATTCTCGCTTACTCGAAGGAAATTATCCCGACACGTCAC GGCTGATTCCTAAAGAATCTAATACGACTGTTGAAATTTCAGCACCCGCACTTTCAGCAGCAATTG AACGGGCATCCCTGTTATCCCACGAAAGTCGCAATAATGTGGTTCGCTTCTCCGTTAACCCAACGG ACAAAACGATTACGATTTTCGGTAACTCACCAGATGTCGGGGAAGTAACGGAACAACTACAGCCCA CTGATTTGTCAGGTGACGAACTTGAAATTTCATTTAACCCAGACTACATGAAAGAAGCGTTACGTT CGTTTGGGCAAGCTATGATCAAAATCTCATTCACGATGGCATTGCGCCCATTCACTTTAGTGCCAA CTGAAGAAGGCGAAAACTTTATTCAGTTGATTACACCAGTTCGAACGTTTTAATTAGCTGAATAAA TATACAATTCTAAATAAAAGGCCCGCATTTCACCGTGAAATGCGGGCCTTTTATGACGGCTAAAAC TAGCTAAAGATGGGGCAATGAATAGGCAATAGCGATGATGTAACTTTTGAAGTTTTAACTTATGCA TATTATATTAGCTGCTGGTGAATTGTCACGCTAACTATCGATGTTGCCCTACTAAAATCCCACCAT CTGATGAACCCCAGTCCTTGCACTCGTTCGCTACAAACGCTCGCTTTCAGATAATGAATGCTGGCT TTAGCCAATTTTTATAGGTAAAAAACAAGATATTTTATTAAAATCCGCAAAATTAGGGAATTTCAG CAAATTTACCCCTAGGCTAGTATCCCGATATTAAAAAGCCCTAAATCGTCTTAAAATGGCTCTGAG TGCGGTTATAAATTGAATTTTCATCAAAA
碱基序列进行比对分析，发现与数据库里的植物乳杆菌序列一致，并保存于武汉市中国典型培养物保藏中心，保藏号为CCTCC NO:M 2017763。
植物乳杆菌FLPL05菌株，是一类厌氧，不产孢子的革兰氏阳性菌，菌体呈短杆状，在MRS琼脂平板培养基中呈乳白色透明，圆形，光滑的菌落。植物乳杆菌FLPL05可以在MRS肉汤培养基上生长，是革兰氏阳性菌。
实施例2：菌株延长小鼠寿命
成年的BALB/c小鼠，随机分两组，每组10只，实验组(FLPL05)：灌胃植物乳杆菌FLPL05至小鼠衰老自然死亡；对照组：灌胃同等体积的生理盐水至小鼠自然死亡。分别统计两组小鼠的自然寿命，并做比较分析。
由图1可以看出，菌株延长小鼠30％的寿命。
实施例3：菌株对老年小鼠肠道渗透性的影响
老年的BALB/c小鼠，随机分两组，每组10只，实验组(FLPL05)：灌胃植物乳杆菌FLPL05；对照组：灌胃同等体积的生理盐水一个月后灌胃 Dextran-FITC，取血清。通过检测血清中Dextran-FITC的含量，评估菌株对老年小鼠肠道渗透性的影响。
由图2可以看出，实验组灌胃植物乳杆菌FLPL05能显著性的改善老年小鼠的肠道渗透性。
实施例4：菌株对老年小鼠肠道结构的影响
灌胃老年鼠一个月后，处死小鼠后取肠道分别做H&E染色。由图3可以看出，菌株对肠道结构有显著的改善作用，与老年小鼠相比，植物乳杆菌 FLPL05干预后，肠绒毛整齐，隐窝完整。
实施例5：菌株干预后降低老年小鼠炎症水平
收集小鼠血清，分别检测血清中炎症指标TNF-α和IFN-γ。
由图4看出，植物乳杆菌FLPL05干预后，能改善炎症水平，血清中的TNF- α降低，IFN-γ增多。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 南昌大学
<120> 一株抗衰老功能的植物乳杆菌FLPL05
<130> 2017
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1477
<212> DNA
<213> Lactobacillus plantarum
<400> 1
acttgacgtt aatactgacg ctattacctt aactggtagt gatgctgata tttctattga 60
aaccaccatt ccagcaagtg atgacaacaa tacgttagtc gttgaagatg ccggttcaat 120
cgttttacct gcccgtttct ttagtgaaat cgtaaaaaaa ttaccagaag acaccatgac 180
tgttaacgtg gtcgacggtt ttcaaacaca aattacttca ggggccgctt cctttacaat 240
caacggctta gaccctgaaa actacccaca cttacctgaa attgatacga ctaataccat 300
cacgttagct ggggatgtac tcaaagagtt gattggtcag accgttattg ccgtttccaa 360
tcaggaaagt cgaccaattc tgaccggggt tcattttatc ttagctaacg gtgagttctt 420
agcggtcgca accgattcac accgcctaag tcaacgccgg atcaagttgc cagaagccaa 480
taatgccaat tatgatgtaa tcattcctgg taaaagtttg acagaattat ctcggatgat 540
cggcgacaat aatcctgacg tgcagatgcg gttatcagaa aaccaagtcc tatttgtgct 600
ggggaatacg tccttctatt ctcgcttact cgaaggaaat tatcccgaca cgtcacggct 660
gattcctaaa gaatctaata cgactgttga aatttcagca cccgcacttt cagcagcaat 720
tgaacgggca tccctgttat cccacgaaag tcgcaataat gtggttcgct tctccgttaa 780
cccaacggac aaaacgatta cgattttcgg taactcacca gatgtcgggg aagtaacgga 840
acaactacag cccactgatt tgtcaggtga cgaacttgaa atttcattta acccagacta 900
catgaaagaa gcgttacgtt cgtttgggca agctatgatc aaaatctcat tcacgatggc 960
attgcgccca ttcactttag tgccaactga agaaggcgaa aactttattc agttgattac 1020
accagttcga acgttttaat tagctgaata aatatacaat tctaaataaa aggcccgcat 1080
ttcaccgtga aatgcgggcc ttttatgacg gctaaaacta gctaaagatg gggcaatgaa 1140
taggcaatag cgatgatgta acttttgaag ttttaactta tgcatattat attagctgct 1200
ggtgaattgt cacgctaact atcgatgttg ccctactaaa atcccaccat ctgatgaacc 1260
ccagtccttg cactcgttcg ctacaaacgc tcgctttcag ataatgaatg ctggctttag 1320
ccaattttta taggtaaaaa acaagatatt ttattaaaat ccgcaaaatt agggaatttc 1380
agcaaattta cccctaggct agtatcccga tattaaaaag ccctaaatcg tcttaaaatg 1440
gctctgagtg cggttataaa ttgaattttc atcaaaa 1477
Claims (6)
1.一株保藏于中国典型培养物保藏中心CCTCC的植物乳杆菌(Lactobacillusplantarum)菌株FLPL05，保藏编号：CCTCC NO:M 2017763。
2.一种微生物菌剂，其特征在于含有权利要求1所述植物乳杆菌菌株FLPL05。
3.根据权利要求2所述的一种微生物菌剂，其特征在于，用于促进健康长寿，保护肠道屏障，降低炎症水平，预防或治疗肠屏障损伤。
4.包含权利要求1所述的植物乳杆菌的乳制品、蔬菜制品、饮料制品和其他发酵类制品。
5.一种组合物，包括权利要求1所述的植物乳杆菌菌株，或通过权利要求2所述的方法所获得的微生物菌剂，或通过权利要求4所述的方法获得的乳制品、蔬菜制品、饮料制品和其他发酵类制品。
6.如权利要求5所述的组合物，其特征在于其是食品，保健品，药品，菌剂。
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