Chimeric endolysin Lys009-Lys009CBD as well as engineering transformation and application thereof

By fusing Lys009CBD to the C-terminus of endolysin Lys009 and then fusing it with the membrane-penetrating peptide P81417, an engineered endolysin P81417-Lys009-Lys009CBD was constructed. This solved the problem of endolysin penetrating the outer membrane of Gram-negative bacteria, achieving highly efficient bactericidal activity and thermal stability against Pseudomonas aeruginosa, and enriching the therapeutic resources against drug-resistant bacteria.

CN122382033AActive Publication Date: 2026-07-14SOUTH CHINA UNIV OF TECH
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Patent Information

Application Number
CN202610837764.9
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-06-11
Publication Date
2026-07-14
Estimated Expiration
2046-06-11

AI Technical Summary

Technical Problem

Existing endosomalins have difficulty effectively penetrating the outer membrane of Gram-negative bacteria, resulting in limited bactericidal effects against Pseudomonas aeruginosa and a risk of drug resistance.

Method used

By fusing its own cell wall binding domain (Lys009CBD) to the C-terminus of the natural endosomalin Lys009, and further fusing it with the membrane-penetrating peptide P81417, an engineered endosomalin P81417-Lys009-Lys009CBD was constructed, achieving efficient membrane penetration and peptidoglycan hydrolysis of Pseudomonas aeruginosa.

Benefits of technology

It significantly improved the peptidoglycan hydrolysis activity against Pseudomonas aeruginosa, reaching 5.30 times that of natural endosomalin, and maintained 100% bactericidal activity even after half an hour at 70°C, providing a highly effective treatment strategy against drug-resistant bacteria.

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Abstract

The invention discloses a chimeric endolysin Lys009-Lys009CBD as well as engineering modification and application thereof, and belongs to the field of gene engineering and protein engineering. According to the invention, the high-activity chimeric endolysin Lys009-Lys009CBD is constructed, and the peptidoglycan hydrolytic activity of the high-activity chimeric endolysin Lys009-Lys009CBD on pseudomonas aeruginosa is 5.30 times that of natural endolysin Lys009, and is 27.98 times that of egg white lysozyme HEWL. The engineering endolysin P81417-Lys009-Lys009CBD is obtained by further fusing a cell-penetrating peptide P81417 to carry out cell-penetrating transformation, and the engineering endolysin can efficiently crack pseudomonas aeruginosa, acinetobacter baumannii and the like and shows excellent thermal stability. According to the invention, a resource library of high-activity endolysin is enriched, important experimental basis and technical support are provided for developing an anti-drug-resistant bacterium treatment strategy, and meanwhile, a new choice is provided for clinically treating multi-drug-resistant bacterium infection.
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