食品科学 ›› 2024, Vol. 45 ›› Issue (2): 195-202.doi: 10.7506/spkx1002-6630-20230505-030

• 生物工程 • 上一篇    下一篇

3 个不同地区母婴粪便来源嗜黏蛋白阿克曼菌的分离及其体外益生特性分析

万开瑞,孟凡姝,陈丽澜,田丰伟,黄丽丽,倪永清   

  1. (1.石河子大学食品学院,新疆 石河子 832003;2.江南大学食品学院,江苏 无锡 214122)
  • 出版日期:2024-01-25 发布日期:2024-02-05
  • 基金资助:
    第八师石河子市科技专项(2020PT01);NSFC-新疆联合基金重点项目(U1903205); 新疆生产建设兵团科技创新团队项目(2020CB007)

Isolation and in Vitro Probiotic Characteristics of Akkermansia muciniphila from Maternal and Infant Feces in Three Different Regions

WAN Kairui, MENG Fanshu, CHEN Lilan, TIAN Fengwei, HUANG Lili, NI Yongqing   

  1. (1. Food College, Shihezi University, Shihezi 832003, China; 2. School of Food Science and Technology, Jiangnan University, Wuxi 214122, China)
  • Online:2024-01-25 Published:2024-02-05

摘要: 本研究采用改良的黏蛋白富集培养基结合实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)检测了48 份母婴粪便样品,优化了分离效果,从8 份阳性样品中分离纯化到24 株嗜黏蛋白阿克曼菌(Akkermansia muciniphila,Akk)。基于16S rRNA基因测序以及Akk菌株特异引物PCR扩增结果,24 株菌全部隶属于Akk。基因外重复回文序列PCR遗传指纹图谱可将24 株菌划分为4 个基因型小群。同时对24 株菌的模拟胃肠液耐受能力、疏水性、抗生素敏感性、聚糖利用能力进行体外检测。结果显示,菌株HN18D-1、HN18D-3、WW48D1-13对模拟胃、肠液耐受性最高。所有Akk分离株对万古霉素、克林霉素、卡那霉素和红霉素具有一定的耐药性,低聚木糖、大豆低聚糖对Akk菌株具有益生元效应。综合以上各项体外益生特性,Akk分离株HN18D-1、HN18D-3、WW48D1-13可作为潜在益生菌菌株进行后续系统的研究。

关键词: 嗜黏蛋白阿克曼菌;16S?rRNA;分离株鉴定;益生特性

Abstract: In this study, a combination of an improved mucin enriched medium with real-time polymerase chain reaction (real-time PCR) was used to test 48 samples of maternal and infant feces for Akkermansia muciniphila (Akk). Under optimized conditions, 24 Akk strains were isolated from eight positive samples. All these strains were confirmed as Akk by 16S rRNA gene sequencing and PCR with Akk-specific primers. Repetitive extragenic palindrome-polymerase chain reaction (rep-PCR) fingerprinting classified the 24 strains into four genotypic groups. Subsequently, these strains were tested in vitro for simulated gastrointestinal fluid tolerance, hydrophobicity, antibiotic susceptibility, and glycan utilization capacity. The results showed that strains HN18D-1, HN18D-3, and WW48D1-13 had the highest tolerance to simulated gastric and intestinal fluids. All Akk strains were resistant to vancomycin, clindamycin, kanamycin and erythromycin. Xylooligosaccharides and soybean oligosaccharides had prebiotic effects on the Akk strains. Collectively, Akk isolates HN18D-1, HN18D-3 and WW48D1-13 can be used as potential probiotic candidates for subsequent in-depth studies.

Key words: Akkermansia muciniphlia; 16S rRNA; isolate identification; probiotic characteristics

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