食品与发酵工业2026，Vol.52Issue(17)：61-73,13.DOI:10.13995/j.cnki.11-1802/ts.046854
生牛乳源假单胞菌蛋白水解特性及3种分型方法对其表型分布一致性的比较研究
Proteolytic properties of Pseudomonas isolated from raw milk and consistency of their phenotypic distribution with three typing methods:A comparative study
摘要
Abstract
Raw milk is prone to quality deterioration during cold storage due to the proteolytic activity of psychrotrophic spoilage bac-teria,particularly those belonging to the genus Pseudomonas.This study was conducted to clarify the proteolytic phenotypic characteristics of raw milk-derived Pseudomonas and to evaluate the applicability of different molecular typing methods in the consistency analysis of their spoilage phenotype clustering.A total of 31 Pseudomonas strains(including 26 isolates from raw milk in Shanghai and 5 type strains)were used as research subjects,and molecular typing was performed based on 16S rRNA gene sequences,ribotyping fingerprints,and a combi-nation of housekeeping genes and protease genes using multi-locus sequence typing-like typing(MLST-like),respectively,to systemati-cally compare the consistency of each typing result with the distribution of proteolytic phenotypes.The results showed that 67.7％of the strains produced clear zones on skim milk agar plates,and the average protease activity of the proteolytic-positive strains was 0.56 △A·h-1·mL-1.In species-level identification,ribotyping exhibited higher discriminatory power than 16S rRNA gene sequencing,with 48.4％of the strains identified as Pseudomonas fluorescens.The proteolytic capacity of the P.fluorescens group was significantly high-er than that of the non-P.fluorescens group(P＜0.000 1).Regarding consistency with the distribution of proteolytic spoilage phenotypes,the clustering results of 16S rRNA gene sequence analysis and ribotyping showed similar overall trends in the distribution of proteolytic phenotypes,while MLST-like typing exhibited the clearest correspondence with proteolytic capacity and the risk classification of proteolytic spoilage.This study revealed the distribution characteristics of proteolytic phenotypes of Pseudomonas in raw milk and evaluated the appli-cability of the three typing methods for identifying spoilage risk.These findings provided a reference for the identification and graded con-trol of spoilage risk posed by Pseudomonas in raw milk.关键词
生牛乳/假单胞菌属/蛋白酶/16S rRNA基因序列分析/核糖体分型/类多位点序列分型Key words
raw milk/Pseudomonas/protease/16S rRNA gene sequence analysis/ribotyping/multi-locus sequence typing-like typing引用本文复制引用
詹翠青,祁岩,游春苹..生牛乳源假单胞菌蛋白水解特性及3种分型方法对其表型分布一致性的比较研究[J].食品与发酵工业,2026,52(17):61-73,13.基金项目
市国资委企业创新发展和能级提升项目(2022013) （2022013）
