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从发酵黄瓜卤汁中检测和鉴定一种裂解性嗜盐四联球菌噬菌体。

Detection and characterization of a lytic Pediococcus bacteriophage from the fermenting cucumber brine.

作者信息

Yoon Sung-Sik, Barrangou-Poueys Roudolphe, Breidt Fred, Fleming Henry P

机构信息

Department of Biological Resources and Technology, Yonsei University, Wonju 220-710, Korea.

出版信息

J Microbiol Biotechnol. 2007 Feb;17(2):262-70.

Abstract

Of the twelve lytic bacteriophages recovered from five different fermenting cucumber tanks that were inoculated with Pediococcus sp. LA0281, a lytic phage, phips05, was characterized in the present study. The plaques were mostly clear and round-shaped on the lawn of starter strain, indicating lytic phage. Overall appearance indicated that it belongs to the Siphoviridae family or Bradley's group B1, with a small isometric head and a flexible noncontractile tail with swollen base plate. The average size was found to be 51.2 nm in head diameter and 11.6 nm wide x 129.6 nm long for the tail. The single-step growth kinetics curve showed that the eclipse and the latent period were 29 min and 34 min, respectively, and an average burst size was calculated to be 12 particles per infective center. The optimum proliferating temperature (35 degrees C) was slightly lower than that of cell growth (35 to 40 degrees C). The structural proteins revealed by SDS-PAGE consisted of one main protein of 33 kDa and three minor proteins of 85, 58, and 52 kDa. The phage genome was a linear double-stranded DNA without cohesive ends. Based on the single and double digestion patterns obtained by EcoRI, HindIII, and SalI, the physical map was constructed. The overall size of the phage genome was estimated to be 24.1 kb. The present report describes the presence of a lytic phage active against a commercial starter culture Pediococcus sp. LA0281 in cucumber fermentation, and a preliminary study characterizes the phage on bacterial successions in the process of starter-added cucumber fermentation.

摘要

从五个接种了乳酸片球菌LA0281(一种裂解性噬菌体)的不同发酵黄瓜罐中回收的12种裂解性噬菌体中,本研究对一种裂解性噬菌体phips05进行了特性分析。在起始菌株的菌苔上,噬菌斑大多清晰且呈圆形,表明是裂解性噬菌体。总体外观表明它属于长尾噬菌体科或布拉德利B1组,具有小的等轴头和基部膨大的柔性非收缩性尾。发现头部直径平均大小为51.2 nm,尾部宽11.6 nm,长129.6 nm。一步生长动力学曲线表明,潜伏期和隐蔽期分别为29分钟和34分钟,计算得出平均裂解量为每个感染中心12个粒子。最佳增殖温度(35℃)略低于细胞生长温度(35至40℃)。SDS-PAGE显示的结构蛋白由一种33 kDa的主要蛋白和三种85、58和52 kDa的次要蛋白组成。噬菌体基因组是无粘性末端的线性双链DNA。基于EcoRI、HindIII和SalI获得的单酶切和双酶切模式构建了物理图谱。噬菌体基因组的总体大小估计为24.1 kb。本报告描述了在黄瓜发酵中存在一种对商业起始培养物乳酸片球菌LA0281有活性的裂解性噬菌体,并对该噬菌体在添加起始菌的黄瓜发酵过程中对细菌演替的影响进行了初步研究。

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