Applied Biochemistry Section, New Zealand Dairy Research Institute, Private Bag 11029, Palmerston North, New Zealand.
Appl Environ Microbiol. 1993 Oct;59(10):3177-82. doi: 10.1128/aem.59.10.3177-3182.1993.
Loosely associated material (LAM) was isolated by gentle extraction procedures from the cell surface of Lactococcus lactis subsp. cremoris E8 and its phage-resistant variant strain 398. LAM from both strains was chemically characterized, and its role in the adsorption of three small isometric bacteriophages, phi 618, phi 833, and phi 852, to the cell surface of the two strains was investigated. The phage-resistant strain (strain 398) produced LAM which differed significantly from the material produced by the parent strain. The total yield of LAM from strain 398 was two- to threefold higher than that from strain E8, and the material contained fivefold more rhamnose and twofold more galactose. Polyacrylamide gel electrophoretic analysis showed that LAM from strain 398 lacked a 21-kDa protein which was present in LAM from the parent strain. Inhibition studies of phage binding by using isolated LAM from two strains showed that although LAM from strain E8 reduced the titer of phi 618 and phi 852 by 53 and 82% respectively, LAM from strain 398 had no effect on the plaque-forming ability of any of the three phages tested. Treatment of LAM from strain E8 with sodium metaperiodate destroyed its ability to bind with phi 618 and phi 852. Phenotypically, strain 398 differed from its parent strain E8 in that it was more prone to cell lysis and required an osmotically adjusted buffer system for the extraction of LAM.
疏松结合物质(LAM)通过温和的提取程序从乳球菌乳亚种。 cremoris E8 及其噬菌体抗性变异株 398 的细胞表面分离出来。对来自两种菌株的 LAM 进行了化学表征,并研究了其在三种小等轴对称噬菌体 phi 618、phi 833 和 phi 852 吸附到两种菌株细胞表面中的作用。噬菌体抗性菌株(菌株 398)产生的 LAM 与亲本菌株产生的物质有明显差异。来自菌株 398 的 LAM 的总产量比来自菌株 E8 的高出两到三倍,并且材料中含有五倍的鼠李糖和两倍的半乳糖。聚丙烯酰胺凝胶电泳分析表明,来自菌株 398 的 LAM 缺乏存在于亲本菌株 LAM 中的 21-kDa 蛋白。使用来自两种菌株的分离 LAM 对噬菌体结合进行抑制研究表明,尽管来自菌株 E8 的 LAM 分别将 phi 618 和 phi 852 的效价降低了 53%和 82%,但来自菌株 398 的 LAM 对测试的三种噬菌体中的任何一种的噬菌斑形成能力均无影响。用高碘酸钠处理来自菌株 E8 的 LAM 会破坏其与 phi 618 和 phi 852 结合的能力。表型上,菌株 398 与其亲本菌株 E8 的不同之处在于它更容易发生细胞裂解,并且需要渗透压调节缓冲系统来提取 LAM。