Design, Screening, and Characterization of Engineered Phage Endolysins with Extracellular Antibacterial Activity against Gram-Negative Bacteria

Sui, Bingrui; Wang, Xinrui; Zhao, Tianyi; Zhen, Jianyu; Ren, Huiying; Liu, Wenhua; Zhang, Xiao-Xuan; Zhang, C

doi:10.1128/aem.00581-23

Cited by 4 publications

(2 citation statements)

References 36 publications

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“…This has been exemplified by the development of artilysins containing positively charged amino acids at one end ( Carratalá et al, 2023 ). Additionally, Sui et al (2023) found that randomly fusing short peptides with endolysin resulted in chimeric endolysins with extracellular activities, characterized by a peptide sequence featuring a positive charge and an α-helical structure. However, the precise differences between ApoE23 and COG133 that account for the divergent activity and lytic spectrum observed between CHU-1 and CHU-2 remain unclear.…”

Section: Discussionmentioning

confidence: 99%

Engineered endolysin of Klebsiella pneumoniae phage is a potent and broad-spectrum bactericidal agent against “ESKAPEE” pathogens

Chen,

Han,

Chen

et al. 2024

Front. Microbiol.

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The rise of antimicrobial resistance in ESKAPEE pathogens poses significant clinical challenges, especially in polymicrobial infections. Bacteriophage-derived endolysins offer promise in combating this crisis, but face practical hurdles. Our study focuses on engineering endolysins from a Klebsiella pneumoniae phage, fusing them with ApoE23 and COG133 peptides. We assessed the resulting chimeric proteins’ bactericidal activity against ESKAPEE pathogens in vitro. ApoE23-Kp84B (CHU-1) reduced over 3 log units of CFU for A. baumannii, E. faecalis, K. pneumoniae within 1 h, while COG133-Kp84B (CHU-2) showed significant efficacy against S. aureus. COG133-L1-Kp84B, with a GS linker insertion in CHU-2, exhibited outstanding bactericidal activity against E. cloacae and P. aeruginosa. Scanning electron microscopy revealed alterations in bacterial morphology after treatment with engineered endolysins. Notably, CHU-1 demonstrated promising anti-biofilm and anti-persister cell activity against A. baumannii and E. faecalis but had limited efficacy in a bacteremia mouse model of their coinfection. Our findings advance the field of endolysin engineering, facilitating the customization of these proteins to target specific bacterial pathogens. This approach holds promise for the development of personalized therapies tailored to combat ESKAPEE infections effectively.

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Section: Discussionmentioning

confidence: 99%

Engineered endolysin of Klebsiella pneumoniae phage is a potent and broad-spectrum bactericidal agent against “ESKAPEE” pathogens

Chen,

Han,

Chen

et al. 2024

Front. Microbiol.

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“…Using the homologous recombination method, Sui et al. ( 2023 ) created a rapid screening platform by combining 20 random peptides to endolysin Bp7e and obtained 6 chimeric endolysins with extracellular antibacterial activity against E. coli , S. enterica , P. aeruginosa , and even S. aureus .…”

Section: Strategy Of Endolysin Against Gram‐negative Bacteriamentioning

confidence: 99%

Engineering strategies and challenges of endolysin as an antibacterial agent against Gram‐negative bacteria

Zheng,

Zhang

2024

Microbial Biotechnology

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Bacteriophage endolysin is a novel antibacterial agent that has attracted much attention in the prevention and control of drug‐resistant bacteria due to its unique mechanism of hydrolysing peptidoglycans. Although endolysin exhibits excellent bactericidal effects on Gram‐positive bacteria, the presence of the outer membrane of Gram‐negative bacteria makes it difficult to lyse them extracellularly, thus limiting their application field. To enhance the extracellular activity of endolysin and facilitate its crossing through the outer membrane of Gram‐negative bacteria, researchers have adopted physical, chemical, and molecular methods. This review summarizes the characterization of endolysin targeting Gram‐negative bacteria, strategies for endolysin modification, and the challenges and future of engineering endolysin against Gram‐negative bacteria in clinical applications, to promote the application of endolysin in the prevention and control of Gram‐negative bacteria.

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Gram-negative endolysins: overcoming the outer membrane obstacle

Sisson,

Jackson,

Fagerlund

et al. 2024

Current Opinion in Microbiology

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Design, Screening, and Characterization of Engineered Phage Endolysins with Extracellular Antibacterial Activity against Gram-Negative Bacteria

Abstract: The presence of the envelope in Gram-negative bacteria limits the use of phage endolysins, and engineering endolysins is an efficient way to optimize their penetrative and antibacterial properties. We built a platform for endolysin engineering and screening.

Cited by 4 publications

References 36 publications

Engineered endolysin of Klebsiella pneumoniae phage is a potent and broad-spectrum bactericidal agent against “ESKAPEE” pathogens

Engineered endolysin of Klebsiella pneumoniae phage is a potent and broad-spectrum bactericidal agent against “ESKAPEE” pathogens

Engineering strategies and challenges of endolysin as an antibacterial agent against Gram‐negative bacteria

Gram-negative endolysins: overcoming the outer membrane obstacle

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