Characterization of staphylococcal endolysin LysSAP33 possessing untypical domain composition

Yu, Jun-Hyeok; Park, Do-Won; Lim, Joong‐Soo; Park, Jong‐Hyun

doi:10.1007/s12275-021-1242-1

Cited by 4 publications

(3 citation statements)

References 38 publications

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“…Notably, the isolated CHAP domain is capable of lysing S. aureus in vivo , with the absence of CBD resulting in a 10-fold decline in enzymatic efficacy. Yu et al engineered a truncated variant of LysSAP33 (residues 1–156; CHAP-156), which shares identity with LysSAP26, albeit originating from a different phage, and observed a significant diminution in lytic efficiency upon removal of the C-terminal domain, underscoring its importance (Yu et al, 2021). Contrary to the findings of Yu et al, our research posits that CHAP proteins lacking the C-terminal domain, exemplified by CHAP SAP26 -161, show superior antimicrobial prowess against a broader spectrum of bacterial strains compared with the native enzyme.…”

Section: Discussionmentioning

confidence: 99%

“…By deleting the C-terminal portion of LysSAP26, we produced two deletion mutants, CHAP SAP26 -139 and CHAP SAP26 -161, and compared their efficacy with the original protein. This decision was guided by prior studies showing the significant role of the CHAP domain in endolysins, such as LysK, and the impact of C-terminal truncations on bactericidal activity, as observed in LysSAP33 (Filatova et al, 2010; Horgan et al, 2009a; Kim et al, 2020b; O’Flaherty et al, 2005; Yu et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

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A novel truncated CHAP modular endolysin, CHAP^SAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

Choi,

Kim,

Dahal

et al. 2024

Preprint

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BackgroundDevelopment of novel antimicrobial agents is imperative due to the increasing threat of antibiotic-resistant pathogens. This study aimed to validate the enhanced antibacterial activity andin vivoefficacy of a novel truncated endolysin, CHAPSAP26-161, derived from the CHAP domain of LysSAP26, against multidrug-resistant bacteria.MethodsTwo deletion mutants, CHAPSAP26-139 and CHAPSAP26-161, were constructed by deleting the C-terminal portion of LysSAP26. These were cloned and expressed, and their antibacterial activities, together with protein purification efficiency, were evaluated against 12 bacterial species under various environmental conditions. To test the temperature and pH stability of the three recombinant proteins, the antibacterial effects of the proteins at various temperatures (4°C–60°C) and pH values (3–10) were measured. Time-kill assay measured the optical density (600 nm) and colony-forming units after incubation for 0, 2, 4, 6, 8, and 24 h. We verified this throughin vivoexperiments using mouse models to evaluate the therapeutic potential of CHAPSAP26-161 againstAcinetobacter baumannii.ResultsCHAPSAP26-161 exhibited higher protein purification efficiency and antibacterial activity than LysSAP26. Moreover, CHAPSAP26-161 showed the highest lytic activity againstA. baumanniiwith a minimal bactericidal concentration (MBC) of 5–10 µg/mL, followed byStaphylococcus aureuswith an MBC of 10–25 µg/mL. Interestingly, CHAPSAP26-161 could lyse anaerobic bacteria, such asC. difficile, with an MBC of 25–50 µg/mL. At pH 4–8 and temperatures of 4°C–45°C, CHAPSAP26-161 exhibited optimal hydrolase activity. The lytic activity of CHAPSAP26-161 was dependent on divalent metal ions, especially Zn2+, and increased in the presence of ethylenediamine tetraacetic acid. CHAPSAP26-161 demonstrated superior protein purification efficiency and antibacterial activity than LysSAP26. It showed high lytic activity against gram-positive, gram-negative, and anaerobic bacteria, includingS. aureusandClostridioides difficile.Enhanced stability under varied temperatures and pH conditions.In vivo,tests demonstrated promising therapeutic effects of CHAPSAP26-161 in murine systemicA. baumanniiinfection models.ConclusionsCHAPSAP26-161, a truncated modular endolysin containing only the CHAP domain of LysSAP26, demonstrated higher protein purification efficiency and antibacterial activity than LysSAP26. It also exhibited extended-spectrum antibacterial activity against gram-positive, gram-negative, and anaerobic bacteria, such asS. aureus,A. baumannii, andC. difficile. Its successfulin vivoapplication in murine models highlights its potential as an alternative therapeutic agent in combating antibiotic resistance.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A novel truncated CHAP modular endolysin, CHAP^SAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

Choi,

Kim,

Dahal

et al. 2024

Preprint

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“…To better understand these complex endolysins derived from phages targeting G + bacteria (mainly including Staphylococcus , Streptococcus , Enterococcus, and Listeria ), we propose a systematic classification of these endolysins based on their domain compositions (Fig. 2 ) and update other types of staphylococcal endolysins given that they were classified into six types [ 41 , 42 ]. The information from the National Center for Biotechnology Information database on the representatives of different types of endolysins is shown in Fig.…”

Section: Introductionmentioning

confidence: 99%

Therapeutic potential of bacteriophage endolysins for infections caused by Gram-positive bacteria

et al. 2023

J Biomed Sci

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Gram-positive (G+) bacterial infection is a great burden to both healthcare and community medical resources. As a result of the increasing prevalence of multidrug-resistant G+ bacteria such as methicillin-resistant Staphylococcusaureus (MRSA), novel antimicrobial agents must urgently be developed for the treatment of infections caused by G+ bacteria. Endolysins are bacteriophage (phage)-encoded enzymes that can specifically hydrolyze the bacterial cell wall and quickly kill bacteria. Bacterial resistance to endolysins is low. Therefore, endolysins are considered promising alternatives for solving the mounting resistance problem. In this review, endolysins derived from phages targeting G+ bacteria were classified based on their structural characteristics. The active mechanisms, efficacy, and advantages of endolysins as antibacterial drug candidates were summarized. Moreover, the remarkable potential of phage endolysins in the treatment of G+ bacterial infections was described. In addition, the safety of endolysins, challenges, and possible solutions were addressed. Notwithstanding the limitations of endolysins, the trends in development indicate that endolysin-based drugs will be approved in the near future. Overall, this review presents crucial information of the current progress involving endolysins as potential therapeutic agents, and it provides a guideline for biomaterial researchers who are devoting themselves to fighting against bacterial infections.

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Molecular Machinery of the Triad Holin, Endolysin, and Spanin: Key Players Orchestrating Bacteriophage-Induced Cell Lysis and their Therapeutic Applications

Samir

2024

PPL

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Phage therapy, a promising alternative to combat multidrug-resistant bacterial infections, harnesses the lytic cycle of bacteriophages to target and eliminate bacteria. Key players in this process are the phage lysis proteins, including holin, endolysin, and spanin, which work synergistically to disrupt the bacterial cell wall and induce lysis. Understanding the structure and function of these proteins is crucial for the development of effective therapies. Recombinant versions of these proteins have been engineered to enhance their stability and efficacy. Recent progress in the field has led to the approval of bacteriophage-based therapeutics as drugs, paving the way for their clinical use. These proteins can be combined in phage cocktails or combined with antibiotics to enhance their activity against bacterial biofilms, a common cause of treatment failure. Animal studies and clinical trials are being conducted to evaluate the safety and efficacy of phage therapy in humans. Overall, phage therapy holds great potential as a valuable tool in the fight against multidrug- resistant bacteria, offering hope for the future of infectious disease treatment.

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Characterization of staphylococcal endolysin LysSAP33 possessing untypical domain composition

Cited by 4 publications

References 38 publications

A novel truncated CHAP modular endolysin, CHAP^SAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

A novel truncated CHAP modular endolysin, CHAP^SAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

Therapeutic potential of bacteriophage endolysins for infections caused by Gram-positive bacteria

Molecular Machinery of the Triad Holin, Endolysin, and Spanin: Key Players Orchestrating Bacteriophage-Induced Cell Lysis and their Therapeutic Applications

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Characterization of staphylococcal endolysin LysSAP33 possessing untypical domain composition

Cited by 4 publications

References 38 publications

A novel truncated CHAP modular endolysin, CHAPSAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

A novel truncated CHAP modular endolysin, CHAPSAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

Therapeutic potential of bacteriophage endolysins for infections caused by Gram-positive bacteria

Molecular Machinery of the Triad Holin, Endolysin, and Spanin: Key Players Orchestrating Bacteriophage-Induced Cell Lysis and their Therapeutic Applications

Contact Info

Product

Resources

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A novel truncated CHAP modular endolysin, CHAP^SAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile

A novel truncated CHAP modular endolysin, CHAP^SAP26-161, that lysesStaphylococcus aureus,Acinetobacter baumannii,andClostridioides difficile