Abstract
To identify and characterize the lysis gene cassette from the bacteriophage ST79 that lyses Burkholderia pseudomallei. Approximately 1·5kb of ST79 lysis genes were identified from the phage genome data. It was composed of holin, peptidase M15A or endolysin, lysB and lysC. Each gene and its combinations were cloned into Escherichia coli and the lytic effects were measured. Co-expression of holin and peptidase M15A showed the highest lysis activity. Expression of holin, lysB/C or holin-peptidase M15A-lysB/lysC lysed the E.coli membrane, whereas peptidase M15A alone did not. The predicted transmembrane structures of holin and lysB/C indicated that they could be inserted into the bacterial membrane to form pores, affecting cell permeability and causing lysis. This is the first report of an investigation into the lysis genes of B.pseudomallei's lytic phage using E.coli as a model. Burkholderia pseudomallei, a Gram-negative bacterium causing an infectious disease, is intrinsically resistant to several antibiotics, and a vaccine is not available. The lysis genes of ST79, the first reported lytic bacteriophage of B.pseudomallei, were characterized. The development of ST79 as an alternative treatment for skin ulceration, for example, or to be used as a gene cloning tool for B.pseudomallei may be possible with this knowledge.
Highlights
A bacteriophage or phage is the virus of bacteria that is very specific to its host
When the MEROPS batch Blast tool was used to detect peptidases and their non-peptidase homologues sequences in ST79 genome, it showed common amino acids among peptidases found in other bacterial genome, such as: B. glumae, B. thailandensis, B. cenocepacia, Burkholderia sp
The increase in antibiotic-resistant bacteria makes the use of possible phage therapy as an alternative treatment for bacterial infections as one of multiple options for treatment
Summary
A bacteriophage or phage is the virus of bacteria that is very specific to its host. It can replicate, multiply after transfection and either lyse the host or become integrated into its genome (Golkar et al 2014). The enzymes related to the lysis mechanism of the phages that are used to lyse the bacterial host during the release of their progeny have been studied and revealed the classical holin-endolysin lysis system found in phages of both Gram-positive and Gram-negative bacteria (Young et al 2000). Endolysins have been considered as a new class of antibiotics as they can destroy the peptidoglycan (PG) of Gram-positive bacterial cell walls. The enzymes or the phages themselves are extensively applied in several fields, for example, the food industry and biological control of unwanted bacteria (Ruyter et al 1997) including pathogenic bacteria in medicine as it shows neither toxicity nor stimulates hyperimmune sera in the mouse model (Jado et al 2003)
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