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Identification and characterization of the two‐component cell lysis cassette encoded by temperate bacteriophage ϕ PYB5 of Lactobacillus fermentum
Author(s) -
Wang S.,
Kong J.,
Zhang X.
Publication year - 2008
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2008.03953.x
Subject(s) - lytic cycle , bacteriophage , lysin , microbiology and biotechnology , biology , lysis , lactobacillus fermentum , temperateness , gene cassette , escherichia coli , virology , bacteria , antibiotics , biochemistry , genetics , gene , antibiotic resistance , virus , integron , lactobacillus plantarum , lactic acid
Aims: To characterize the two‐component cell lysis cassette comprised of holin (Hyb5) and endolysin (Lyb5) encoded by Lactobacillus fermentum temperate bacteriophage ϕ PYB5, and illustrate the potential application of Lyb5 as therapeutic agents. Methods and Results: The hyb5–lyb5 cassette was cloned from the genome library of ϕ PYB5, and the hyb5 , lyb5 and hyb5–lyb5 cassette were expressed in E. coli BL21, respectively. The molecular weight of Hyb5 indicated by SDS‐PAGE was 19 kDa, and Lyb5 was 45 kDa. Both Hyb5 and Lyb5 protein could induce cell lysis alone, resulting in the leakage of β ‐galactosidase. However, the Hyb5–Lyb5 cassette lysed the host cells more rapidly and extensively. By zymogram analysis, Lyb5 exhibited a broad lytic spectrum. Conclusions: Overexpression of hyb5 , lyb5 and hyb5 – lyb5 cassette were carried out in E. coli and Lyb5 exhibited a broad lytic spectrum. Significance and Impact of the Study: The Lyb5 produced in E. coli exhibited a broad lytic spectrum against Gram‐positive strains including Staphylococcus aureus as well as Gram‐negative strains such as Salmonella typhi , suggesting that Lyb5 provides a potential alternative of diagnostic tools and therapeutic agents.