전지환 (성균관대학교)

한빛사논문

조회139

전지환Jihwan Chun

성균관대학교

Int. J. Antimicrob. Agents, May 23 2024, S0924-8579(24)00134-1 | https://doi.org/10.1016/j.ijantimicag.2024.107216 Delivery of endolysin across outer membrane of Gram-negative bacteria using translocation domain of botulinum neurotoxin

Wonbeom Park ^a†, Myungseo Park^a,b†, Jihwan Chun ^a†, Jaehyeon Hwang ^a, Suhyun Kim ^a, Nayoon Choi ^a, Sumin Kim ^a, Seongju Kim ^a, Sangwon Jung ^c, Kwan Soo Ko ^d, Dae-Hyuk Kweon ^a*

^aDepartment of Integrative Biotechnology, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 16419, Republic of Korea

^bInfectious Diseases Therapeutic Research Center, Korea Research Institute of Chemical Technology (KRICT), Daejeon 34114, Republic of Korea

^cResearch Center, MVRIX, Anyang 14058, Republic of Korea

^dDepartment of Microbiology, Sungkyunkwan University School of Medicine, Suwon, Republic of Korea

^†These authors contributed equally to this work and share first authorship.

^*Corresponding author: Dae-Hyuk Kweon, PhD

Abstract

The emergence of multidrug resistant pathogens has outpaced the development of new antibiotics, leading to renewed interest in endolysins. Endolysins have been investigated as novel biocontrol agents for Gram-positive bacteria. However, their efficacy against Gram-negative species is limited by the barrier presented by their outer membrane, which prevents endolysin access to the peptidoglycan substrate. Here, we used the translocation domain of botulinum neurotoxin (BoNT) to deliver endolysin across the outer membrane of Gram-negative bacteria. The translocation domain selectively interacts with and penetrates membranes composed of anionic lipids, which have been used in nature to deliver various proteins into animal cells. In addition to the BoNT translocation domain, we have fused bacteriophage-derived receptor binding protein to endolysins. This allows the attached protein to efficiently bind to a broad spectrum of Gram-negative bacteria. By attaching these target-binding and translocation machinery to endolysins, we aimed to develop an engineered endolysin with broad-spectrum targeting and enhanced antibacterial activity against Gram-negative species. To validate our strategy, we designed engineered endolysins using two well-known endolysins, T5 and LysPA26, and tested them against 23 strains from six species of Gram-negative bacteria, confirming that our machinery can act broadly. In particular, we observed a 2.32 log reduction in 30 minutes with only 0.5 µM against an A. baumannii isolate. We also used the SpyTag/SpyCatcher system to easily attach target-binding proteins, thereby improving its target binding ability. Overall, our newly developed endolysin engineering strategy may be a promising approach to control multidrug-resistant Gram-negative bacterial strains.

논문정보

형식Research article
게재일2024년 05월 (BRIC 등록일 2024-05-28)
연구진국내 연구진
분야 생명과학 > 생물공학

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