Deveau Hélène, Van Calsteren Marie-Rose, Moineau Sylvain
Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie, Groupe de Recherche en Ecologie Buccale (GREB), Faculté de Médecine Dentaire, Université Laval, Québec, Canada G1K 7P4.
Appl Environ Microbiol. 2002 Sep;68(9):4364-9. doi: 10.1128/AEM.68.9.4364-4369.2002.
In this study, we report that Lactococcus lactis strains producing exopolysaccharides (EPS) are sensitive to virulent phages. Eight distinct lytic phages (Q61 to Q68) specifically infecting Eps(+) strains were isolated in 47 buttermilk samples obtained from 13 North American factories. The eight phages were classified within the 936 species by the multiplex PCR method, indicating that these phages are not fundamentally distinct from those infecting Eps(-) L. lactis strains. The host range of these phages was determined with 19 Lactococcus strains, including 7 Eps(+) and 12 Eps(-) cultures. Three phages (Q62, Q63, and Q64) attacked only the Eps(+) strain SMQ-419, whereas the five other phages (Q61, Q65, Q66, Q67, and Q68) infected only the Eps(+) strain SMQ-420. The five other Eps(+) strains (H414, MLT2, MLT3, SMQ-461, and SMQ-575) as well as the 12 Eps(-) strains were insensitive to these phages. The monosaccharide composition of the polymer produced by the seven Eps(+) strains was determined. The EPS produced by strains MLT3, SMQ-419, and SMQ-575 contained glucose, galactose, and rhamnose. The EPS fabricated by H414 contained only galactose. The EPS made by MLT2, SMQ-420, and SMQ-461 contained glucose and galactose. These findings indicate that the sugar composition of the EPS has no effect on phage sensitivity. The plasmid encoding the eps operon was cured from the two phage-sensitive strains. The cured derivatives were still phage sensitive, which indicates that EPS are not necessary for phage infection. Phage adsorption assays showed that the production of EPS does not confer a significant phage resistance phenotype.
在本研究中,我们报告产胞外多糖(EPS)的乳酸乳球菌菌株对烈性噬菌体敏感。从北美13家工厂获得的47份酪乳样品中分离出8种特异性感染Eps(+)菌株的不同裂解性噬菌体(Q61至Q68)。通过多重PCR方法将这8种噬菌体归类于936种内,表明这些噬菌体与感染Eps(-)乳酸乳球菌菌株的噬菌体并无本质区别。用19株乳酸乳球菌菌株(包括7株Eps(+)和12株Eps(-)培养物)测定了这些噬菌体的宿主范围。三种噬菌体(Q62、Q63和Q64)仅攻击Eps(+)菌株SMQ - 419,而其他五种噬菌体(Q61、Q65、Q66、Q67和Q68)仅感染Eps(+)菌株SMQ - 420。其他五种Eps(+)菌株(H414、MLT2、MLT3、SMQ - 461和SMQ - 575)以及12株Eps(-)菌株对这些噬菌体不敏感。测定了7株Eps(+)菌株产生的聚合物的单糖组成。MLT3、SMQ - 419和SMQ - 575菌株产生的EPS含有葡萄糖、半乳糖和鼠李糖。H414产生的EPS仅含有半乳糖。MLT2、SMQ - 420和SMQ - 461产生的EPS含有葡萄糖和半乳糖。这些发现表明EPS的糖组成对噬菌体敏感性没有影响。从两株噬菌体敏感菌株中消除了编码eps操纵子的质粒。消除后的衍生物仍然对噬菌体敏感,这表明EPS对于噬菌体感染不是必需的。噬菌体吸附试验表明EPS的产生不会赋予显著的噬菌体抗性表型。