Complete Genome of a Novel Lytic Vibrio parahaemolyticus Phage VPp1 and Characterization of Its Endolysin for Antibacterial Activities
Complete Genome of a Novel Lytic Vibrio parahaemolyticus Phage VPp1 and Characterization of Its Endolysin for Antibacterial Activities
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DOI：
10.4315/0362-028x.jfp-17-278
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发表时间：
2018-07-01
影响因子：
2
通讯作者：
Jiang, Xiuping
中科院分区：
文献类型：
--
作者：
Li, Mengzhe;Jin, Yanqiu;Jiang, Xiuping
副溶血性弧菌是一种重要的食源性病原体，通常通过生的或未煮熟的海鲜传播。来源于噬菌体的内溶素为控制细菌病原体提供了新的途径。本研究的目的是对一种新的溶菌性副溶血弧菌噬菌体VPp 1进行测序，并测定该噬菌体衍生的重组内溶素（LysVPp 1）的抗菌活性。VPp 1基因组全长为50, 431 bp，G+C含量为41.35%。该基因组编码67个开放阅读框（ORF），包括核苷酸代谢、复制、结构、包装、裂解和其他功能。两个tRNA编码携带反密码子UGG和CCA。在这些功能蛋白中，ORF 33被推断为编码内溶素，而在VPp 1基因组中没有发现holin/antiholin或Rz/Rz 1裂解基因等价物。ORF 33的克隆和表达。内溶素LysVPp 1能裂解12株副溶血弧菌中的9株，显示其比噬菌体VPp 1具有相对更广的宿主谱，噬菌体VPp 1仅裂解12株副溶血弧菌中的3株。此外，对于EDTA预处理的细菌细胞，LysVPp 1处理组在450 nm处的光密度降低了0.4，而对照组的光密度小于0.1，这表明水解性能增强。这些结果有助于开发用于控制副溶血性弧菌的新型酶制剂的潜力。
Vibrio parahaemolyticus is an important foodborne pathogen that is generally transmitted via raw or undercooked seafood. Endolysins originating from bacteriophages offer a new way to control bacterial pathogens. The objectives of this study were to sequence a novel lytic V. parahaemolyticus phage VPp1 and determine the antibacterial activities of the recombinant endolysin (LysVPp1) derived from this phage. The complete VPp1 genome contained a double-stranded DNA of 50,431 bp with a total G+C content of 41.35%. The genome was predicted to encode 67 open reading frames (ORFs), which were organized as nucleotide metabolism, replication, structure, packaging, lysis, and some additional functions. Two tRNAs were encoded to carry anticodons UGG and CCA. Among the functional proteins, ORF33 was deduced to encode endolysin, whereas no holin/antiholin or Rz/Rz1 lysis gene equivalents were found in the VPp1 genome. ORF33 was cloned and expressed. The endolysin LysVPp1 could lyse 9 of 12 V. parahaemolyticus strains, showing its relatively broader host spectrum than phage VPp1, which lysed only 3 of 12 V. parahaemolyticus strains. Furthermore, for EDTA-pretreated bacterial cells, the optical density of the LysVPp1 treatment group decreased by 0.4 at 450 nm, compared with less than 0.1 in control groups, demonstrating enhanced hydrolytic properties. These results contribute to the potential for development of novel enzybiotics for controlling V. parahaemolyticus.