食品与发酵工业 >

2023 , Vol. 49 >Issue 19: 67 - 74

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.034801

研究报告

一株分离自母乳的长双歧杆菌婴儿亚种YLGB-1496的菌株鉴定

  • 刘冲 ,
  • 马霞 ,
  • 于学健 ,
  • 刘艺茹 ,
  • 刘伟贤 ,
  • 刘蕊 ,
  • 辛迪 ,
  • 唐腾飞 ,
  • 刘红强 ,
  • 葛媛媛 ,
  • 孙婷 ,
  • 蒋秋悦 ,
  • 洪维鍊 ,
  • 姚粟
展开
  • 1(中国食品发酵工业研究院有限公司 中国工业微生物菌种保藏管理中心,北京,100015)
    2(内蒙古乳业技术研究院有限责任公司,内蒙古 呼和浩特,010110)
    3(内蒙古伊利实业集团股份有限公司,内蒙古 呼和浩特,010110)
第一作者:硕士(姚粟正高级工程师和洪维鍊博士为共同通信作者,E-mail:milly@china-cicc.org;hongweilian@yili.com)

收稿日期: 2023-01-03

  修回日期: 2023-02-07

  网络出版日期: 2023-11-01

收起

Strain identification of Bifidobacterium longum subsp. infantis YLGB-1496 isolated from breast milk

  • LIU Chong ,
  • MA Xia ,
  • YU Xuejian ,
  • LIU Yiru ,
  • LIU Wei-hsien ,
  • LIU Rui ,
  • XIN Di ,
  • TANG Tengfei ,
  • LIU Hongqiang ,
  • GE Yuanyuan ,
  • SUN Ting ,
  • JIANG Qiuyue ,
  • HUNG Wei-Lian ,
  • YAO Su
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  • 1(China National Research Institute of Food and Fermentation Industries Co.LTD., China Center of Industrial Culture Collection, Beijing 100015, China)
    2(Inner Mongolia Dairy Technology Research Institute Co.Ltd., Hohhot 010110, China)
    3(Inner Mongolia Yili Industrial Group Co.Ltd., Hohhot 010110, China)

Received date: 2023-01-03

  Revised date: 2023-02-07

  Online published: 2023-11-01

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摘要

以母乳来源的长双歧杆菌婴儿亚种YLGB-1496为研究对象,收集5株不同批次的YLGB-1496作为目标菌株,收集8株长双歧杆菌参比菌株实物,同时收集28株长双歧杆菌婴儿亚种全基因组序列。采用形态学、生理生化、基质辅助激光解析电离飞行时间质谱鉴定技术和全基因组测序技术,建立了适用于YLGB-1496的菌株鉴定方法。结果表明,5株YLGB-1496与模式菌株B.longum subsp. infantis ATCC 15697T的平均核苷酸一致性值均大于98％,数字DNA-DNA杂交值均大于85%,5株不同批次来源YLGB-1496鉴定为长双歧杆菌婴儿亚种。对YLGB-1496和参比菌株的1 059个共有核心基因开展多位点序列分型分析,结果表明该菌株可与其他参比菌株有效区分。以菌株YLGB-1 496-1的基因组序列为参考开展单核苷酸位点多态性分析,结果表明YLGB-1496不同来源菌株间SNP差异小于30,并且与其他参比菌株差异大于18 000,系统发育分析可以有效区分YLGB-1496与其他参比菌株。益生菌的安全性和功能性评价均在菌株水平具有特异性,研究建立的表型、基因型相结合的菌株鉴定方法可为菌株鉴定标准提供有效技术支撑,对促进益生菌在食品行业更加安全和广泛的应用具有重要意义。

关键词： 长双歧杆菌婴儿亚种; 菌株鉴定; 全基因组测序; 多位点序列分型; 单核苷酸位点多态性

本文引用格式

刘冲 , 马霞 , 于学健 , 刘艺茹 , 刘伟贤 , 刘蕊 , 辛迪 , 唐腾飞 , 刘红强 , 葛媛媛 , 孙婷 , 蒋秋悦 , 洪维鍊 , 姚粟 . 一株分离自母乳的长双歧杆菌婴儿亚种YLGB-1496的菌株鉴定[J]. 食品与发酵工业, 2023 , 49(19) : 67 -74 . DOI: 10.13995/j.cnki.11-1802/ts.034801

Abstract

Bifidobacterium longum subsp. infantis YLGB-1496 isolated from breast milk was studied in this paper. Five strains of YLGB-1496 from different batches were collected as target strains, and eight strains of B. longum were collected as the references, and other 28 complete genomes of B. longum subsp. infantis were collected as references. Microbial identification technologies including morphology, physiological and biochemical analysis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and whole genome sequencing were applied to establish the strain-level identification method suitable for the identification of strain YLGB-1496. The results showed that the average nucleotide identity values of five strains of YLGB-1496 were greater than 98%, and the digital DDH values were all greater than 85% comparing with the type strain B. longum subsp. infantis ATCC 15697T. Five strains of YLGB-1496 from different sources were identified as B. longum subsp. infantis. Complete genome multilocus sequence typing (cgMLST) analysis was carried out based on the 1 059 core genes of the strains of YLGB-1496 and other reference strains, the results showed that the strains YLGB-1496 could be effectively distinguished from other reference strains. Single nucleotide polymorphism (SNP) analysis was carried out based on the whole genome sequence of the strain YLGB-1496-1, and results showed that the difference in SNP between different sources of YLGB-1496 was less than 30, and greater than 18 000 between reference strains. Phylogenetic analysis further demonstrated that the SNP analysis can effectively distinguish YLGB-1496 from other reference strains. The safety and function of probiotics were specific at the strain level. The strain-level identification method that combines phenotype and genotype established in this study could provide a technical basis for the establishment of strain identification relative standards and promote the safer and wider application of probiotics in the food industry.

Key words： Bifidobacterium longum subsp. infantis; strain identification; whole genome sequencing; complete genome multilocus sequence typing (cgMLST); single nucleotide polymorphism (SNP)

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