以婴儿和成人肠道来源的菌株K56和ET-22为研究对象,选取8株其他来源菌株作为参考,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、全基因组测序、多位点序列分型分析和单核苷酸多态性(single nucleotide polymophism,SNP)分析,建立了适用于菌株K56和ET-22的鉴定方法。结果表明,菌株K56和ET-22及参考菌株的基质辅助激光解吸/电离飞行时间鉴定比对值大于2,平均核苷酸一致性值(average nucleotide identity,ANI)达到95％以上,在种水平均鉴定为副干酪乳杆菌(Lactobacillus paracasei);对副干酪乳杆菌K56、ET-22和参考菌株的1 701个共有核心基因开展基于全基因组测序的多基因序列位点分析,菌株K56和ET-22可与其他参考菌株有效区分,SNP分析表明不同培养代数间副干酪乳杆菌K56的SNP差异为103、ET-22的SNP差异为49,系统发育分析表明该方法可以有效区分副干酪乳杆菌K56、ET-22与其他参考菌株。益生菌的安全性和功能性均在菌株水平具有特异性,已成为业内新的科学共识。该研究建立的益生菌K56和ET-22菌株水平精准鉴定方法体系,对促进其在乳品行业的广泛应用具有重要意义。
刘艺茹
,
马霞
,
赵婷
,
刘锦浲
,
于学健
,
刘伟贤
,
程坤
,
张欣
,
曹艳花
,
辛迪
,
冯慧军
,
刘福东
,
赵雯
,
洪维鍊
,
姚粟
. 分离自人体肠道的副干酪乳杆菌K56和ET-22的菌株鉴定[J]. 食品与发酵工业, 2022
, 48(1)
: 62
-69
.
DOI: 10.13995/j.cnki.11-1802/ts.026255
Strain K56 and ET-22 isolated from the infant intestine were studied in this paper, and the other eight strains of Lactobacillus spp.isolated from different sources were collected as the reference. Microbial identification techniques including morphology, physiological and biochemical analysis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), whole genome sequencing, whole genome multilocus sequence typing(wgMLST) and whole genome single nucleotide polymorphism (SNP) were applied to establish the identification method for strain K56 and ET-22.K56, ET-22, and other eight strains of Lactobacillus spp.isolated from different sources were studied. The MALDI-TOF identification scores of strain K56 and ET-22 were greater than 2, and the average nucleotide identity (ANI) values were greater than 95% comparing with the type strain, which indicated strain K56 and ET-22 were identified as Lactobacillus paracasei. wgMLST analysis was carried out based on the 1 701 core genes of the L. paracasei K56, L. paracasei ET-22 and other 8 reference strains, the results showed that the strain K56 and ET-22 could be effectively distinguished from other reference strains. SNP analysis results showed that the difference in SNP between different generations of L. paracasei K56 and L. paracasei ET-22 were 103 and 49 respectively, and the phylogenetic analysis further demonstrated that the SNP analysis can effectively distinguish L. paracasei K56 and L. paracasei ET-22 from other reference strains. The safety and functional properties of probiotics are specific at the strain level, which has become a scientific consensus. The accurate identification method system researched for probiotics K56 and ET-22 strains is helpful to promote their application in the dairy industry.
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