标题:Functional analysis of the N-terminal region of endolysin Lyb5 encoded by Lactobacillus fermentum bacteriophage phi PYB5
作者:Guo, Tingting; Zhang, Chenchen; Liu, Wei; Wang, Shaohua; Kong, Jian
作者机构:[Guo, Tingting; Zhang, Chenchen; Liu, Wei; Wang, Shaohua; Kong, Jian] Shandong Univ, State Key Lab Microbial Technol, Jinan 250100, Peoples R China.; 更多
通讯作者:Kong, J
通讯作者地址:[Kong, J]27 Shanda Nanlu, Jinan 250100, Peoples R China.
来源:INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
出版年:2015
卷:203
页码:1-7
DOI:10.1016/j.ijfoodmicro.2015.02.033
关键词:Lactobacillus fermentum; Bacteriophage; Endolysin; N-terminal;; Signal-anchor-release domain
摘要:Lactobacillus fermentum temperate bactenophage phi PYB5 uses endolysin Lyb5 and holin Hyb5 to burst the host cell. Previous results showed that expression of Lyb5 in Escherichia colt caused host cell lysis slowly, leading us to suppose that Lyb5 could pass the cytoplasmic membrane partly. In this work, the function of a putative signal peptide (SPLyb5) at the N-terminal of Lyb5 was investigated. In E. coli, the cell adopted a spherical shape during induction of Lyb5 protein, while morphological changes were not observed during expression of the SPLyb5 truncation, indicating that the SPLyb5 motif may serve as a functional signal peptide. However, SPLyb5 was not proteolytically cleaved at the predicted site during the translocation of Lyb5, and the expressed Lyb5 protein appeared in the cytoplasm, cytoplasmic membrane and periplasm fractions with the same molecular mass. Similar results were obtained using Lactococcus lactis as a host to express Lyb5. These results indicated that SPLyb5 could direct Lyb5 to the periplasm in a membrane-tethered form, and then release it as a soluble active enzyme into the periplasm. In addition, SPLyb5 could also drive the fused NucleaseB protein to the extracytoplasm environment in E. colt as well as in L. lactis. We proposed that in Gram-negative and Gram-positive hosts SPLyb5 acted as a signal-anchor-release domain, which was firstly identified here by experimental evidences in lactic acid bacteria phages. The application of signal-anchor-release domain for endolysin export in bacteriophages infecting Gram-positive and Gram-negative hosts was discussed. (C) 2015 Elsevier B.V. All rights reserved.
收录类别:SCIE
WOS核心被引频次:3
资源类型:期刊论文
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