title: The antibacterial activity of E. coli bacteriophage lysin lysep3 is enhanced by fusing the Bacillus amyloliquefaciens bacteriophage endolysin binding domain D8 to the C-terminal region.
abstract: Bacteriophage endolysin is one of the most promising antibiotic substitutes, but in Gram-negative bacteria, the outer membrane prevents the lysin from hydrolyzing peptidoglycans and blocks the development of lysin applications. The prime strategy for new antibiotic substitutes is allowing lysin to access the peptidoglycan from outside of the bacteria by reformation of the lysin. In this study, the novel Escherichia coli (E. coli) phage lyase lysep3, which lacks outside-in catalytic ability, was fused with the N-terminal region of the Bacillus amyloliquefaciens lysin including its cell wall binding domain D8 through the best manner of protein fusion based on the predicted tertiary structure of lysep3-D8 to obtain an engineered lysin that can lyse bacteria from the outside. Our results showed that lysep3-D8 could lyse both Gramnegative and Gram-positive bacteria, whereas lysep3 and D8 have no impact on bacterial growth. The MIC of lysep3-D8 on E. coli CVCC1418 is 60 μg/ml; lysep3-D8 can inhibit the growth of bacteria up to 12 h at this concentration. The bactericidal spectrum of lysep3-D8 is broad, as it can lyse of all of 14 E. coli strains, 3 P. aeruginosa strains, 1 Acinetobacter baumannii strain, and 1 Streptococcus strain. Lysep3-D8 has sufficient bactericidal effects on the 14 E. coli strains tested at the concentration of 100 μg/ml. The cell wall binding domain of the engineered lysin can destroy the integrity of the outer membrane of bacteria, thus allowing the catalytic domain to reach its target, peptidoglycan, to lyse the bacteria. Lysep3-D8 can be used as a preservative in fodder to benefit the health of animals. The method we used here proved to be a successful exploration of the reformation of phage lysin.
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Jingmin
authors.initials: J
authors.lastname: Gu
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Meng
authors.initials: M
authors.lastname: Lv
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Guo
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Guangmou
authors.initials: G
authors.lastname: Yan
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Yu
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Chongtao
authors.initials: C
authors.lastname: Du
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Feng
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Han
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China. schangjiang@126.com.
authors.forename: Changjiang
authors.initials: C
authors.lastname: Sun
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, Jilin, 130062, P. R. China. leiliancheng@163.com.
authors.forename: Liancheng
authors.initials: L
authors.lastname: Lei
mesh_terms: D040981:Acinetobacter baumannii / Q000187:drug effects; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D017103:Bacillus Phages / Q000737:chemistry*; D000069977:Bacillus amyloliquefaciens / Q000235:genetics*; D001433:Bacteriolysis; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism; D003090:Coliphages / Q000737:chemistry*; D010450:Endopeptidases / Q000235:genetics* / Q000494:pharmacology*; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D010457:Peptidoglycan / Q000187:drug effects; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D014764:Viral Proteins / Q000235:genetics / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D010457:Peptidoglycan; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D010450:Endopeptidases; C001143:endolysin
keywords: antibacterial activity; endolysin binding domain D8; endolysin lysep3; fusing expression