Functional analysis of antibacterial activity of Bacillus amyloliquefaciens phage endolysin against Gram‐negative bacteria

Abstract

To analyze the antibacterial activity of Bacillus amyloliquefaciens phage endolysin, nine deletion derivatives of the endolysin were constructed. Each deletion mutant was overexpressed, purified and characterized. The catalytic domain was located on the N-terminal region and the C-terminus had an affinity with the bacterial envelope. The enzymatic activity remained in spite of the deletion of the C-terminal 116-amino acid region; however, the antibacterial activity was lost. These results indicate that antibacterial action requires both the C-terminal cell-binding and the N-terminal enzymatic activities. ß

Molecular Aspects and Comparative Genomics of Bacteriophage Endolysins

“…lactis bacteriophage LL-H endolysin can be removed without destroying the lytic activity (85). C-terminal deletions of the Staphylococcus aureus bacteriophage endolysins PlyTW and Ply187, L. monocytogenes bacteriophage endolysin Ply511, Bacillus amyloliquefaciens phage endolysin Morita2001, Bacillus anthracis prophage endolysin PlyL, and Bacillus cereus phage endolysin Ply21 even increase the muralytic activity (50,83,(86)(87)(88). This pattern is also present in endolysins with dual lytic domains, for which, in theory, both should be equally active.…”

Characterization and genome sequencing of a Citrobacter freundii phage CfP1 harboring a lysin active against multidrug-resistant isolates

“…On the other hand, the activity of the CfP1 phage seems to tolerate higher pH values better than the recombinant lysin, which indicates that other lytic accessory proteins might have an important role in phage antibacterial activity. The ability of a Gram-negative-specific phage lysin to kill cells Bfrom without^has been solely attributed to few enzymes (e.g., Enterobacter phage T4, Bacillus phage PlyL, Acinetobacter phage PlyF307, and Pseudmonas phage OBP lysins) (During et al 1999;Morita et al 2001;Walmagh et al 2012;Lood et al 2015). This effect was associated to the positively charged amino acids located at the enzyme's Ctermini, which is able to disrupt or mediate lysin access to the peptidoglycan, causing subsequent bacteriolysis.…”

Bacteriophage T4 Genome

“…Even today, one of the best sources of T-even-like phages is the stools of patients recovering from dysentery (L. Gachechiladze and H. Brussow, personal communication). Specific proteins, phage lysins in particular, have been proposed as useful enzymes for killing troublesome bacteria (66,759). Recently, purified PlyG lysin (an N-acetylmuramoyl-L-alanine amidase), produced by gamma phage of Bacillus anthracis, was shown to effectively kill the bacterium (967).…”