Design SMAP29-LysPA26 as a Highly Efficient Artilysin against Pseudomonas aeruginosa with Bactericidal and Antibiofilm Activity.
Design SMAP29-LysPA26 as a Highly Efficient Artilysin against Pseudomonas aeruginosa with Bactericidal and Antibiofilm Activity.
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DOI：
10.1128/spectrum.00546-21
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发表时间：
2021-12-22
影响因子：
3.7
通讯作者：
Zhang C
中科院分区：
文献类型：
--
作者：
Wang T;Zheng Y;Dai J;Zhou J;Yu R;Zhang C
抗生素耐药性（AMR）是全球健康的主要问题。多重耐药革兰阴性菌感染，特别是耐碳青霉烯类病原菌感染，迫切需要有效的抗生素和新的治疗方法。然而，针对革兰氏阴性菌的创新方法的科学挑战阻碍了抗生素药物的研究和开发。噬菌体来源的内溶素是溶菌的，并且对细菌种或属具有特异性，提供了有希望的抗生素策略。而革兰氏阴性菌的外膜可以阻止肽聚糖层被内溶素水解。抗菌肽通常会破坏外膜的稳定性，从而增强内溶素的抗菌活性。在本研究中，我们设计了在LysPA 26的N-末端融合抗菌肽SMAP 29的新的artilysins（命名为AL-3AA、AL-9AA和AL-15 AA），并对它们进行了评价。结果表明，关节溶素AL-3AA是高度杀菌的;即使0.05 mg/mL AL-3AA也可以在60 min内在不含EDTA的情况下减少5.81 log单位的铜绿假单胞菌。它通过细胞裂解快速且剂量依赖性地杀死铜绿假单胞菌。AL-3AA抑制铜绿假单胞菌PAO 1生物膜形成，并显著减少成熟的铜绿假单胞菌生物膜。它也有潜在的广谱活性，对敏感的革兰氏阴性菌在医院，包括K。pneumoniae和E.杆菌抗菌机理的研究为AL-3AA的抗菌作用提供了有价值的信息，AL-3AA能迅速溶解和分解细菌。这些结果表明AL-3AA可能是一种新的和有前途的抗铜绿假单胞菌的抗菌剂。抗菌药物耐药性（Antimicrobial resistance，AMR）是全球健康面临的重大问题，尤其是革兰阴性菌的多重耐药（multidrug resistant，MDR）感染，更是对人类健康带来巨大挑战。即使是新的抗生素研究正在进行中，临床上用于治疗革兰氏阴性菌的抗生素也仅限于一小部分分子支架，迫切需要生物分子类别的抗生素。在这项研究中，我们设计了新的蛋白结合抗菌肽和内溶素协同杀菌作用。AL-3AA蛋白具有很强的杀菌活性，能通过溶解铜绿假单胞菌而迅速杀死铜绿假单胞菌，并呈剂量依赖性。它还杀死了肺炎克雷伯菌和大肠杆菌，显示出对医院中敏感的革兰氏阴性菌的潜在广谱活性。AL-3AA有望成为一种新的抗铜绿假单胞菌的抗菌药物。
Antimicrobial resistance (AMR) is a major issue to global health. The multidrug-resistant (MDR) Gram-negative infections, particularly infected by carbapenem-resistant pathogens, urgently need efficient antibiotics and novel therapy. However, the scientific challenges of aiming for innovative approaches against Gram-negative bacteria have hindered the research and development of antibiotic drugs. Phage-derived endolysins are bacteriolytic and specific for a bacterial species or genus, providing a promising antibiotic strategy. However, the outer membrane of Gram-negative bacteria could prevent the peptidoglycan layer from the hydrolysis of endolysins. Antimicrobial peptides usually destabilize the outer membrane and could enhance the antibiotic activity of endolysins. In this study, we designed new artilysins with antimicrobial-peptide SMAP29 fusion at the N-terminal of LysPA26 (named as AL-3AA, AL-9AA, and AL-15AA), and evaluated them. The results showed artilysin AL-3AA to be highly bactericidal; even 0.05 mg/mL AL-3AA could reduce 5.81 log units P. aeruginosa without EDTA in 60 min. It killed P. aeruginosa rapidly and dose-dependently through cell lysis. AL-3AA inhibited P. aeruginosa PAO1 biofilm formation and significantly decreased mature P. aeruginosa biofilms. It also had potential broad-spectrum activity against susceptible Gram-negative bacteria in the hospital, including K. pneumoniae and E. coli. The antibacterial mechanism investigation has provided valuable information about the antibacterial action of AL-3AA, which can lyse and disintegrate the bacterial quickly. These results suggested AL-3AA could be a new and promising antimicrobial agent for the combat of P. aeruginosa. IMPORTANCE Antimicrobial resistance (AMR) is a major issue to global health, particularly the multidrug-resistant (MDR) Gram-negative infections, which pose great challenges. Even new antibiotics research is ongoing, antibiotics used to treat Gram-negative bacteria in the clinical are limited in a small set of molecular scaffolds, and biomolecular categories of antibiotics are urgently needed. In this study, we designed new proteins by combining antimicrobial peptides and endolysins for synergistic bactericidal effects. One of designed proteins, named AL-3AA, showed highly bactericidal, and killed P. aeruginosa rapidly and dose-dependently through cell lysis. It also killed Klebsiella pneumoniae and Escherichia coli, showing potential broad-spectrum activity against susceptible Gram-negative bacteria in the hospital. All results suggest AL-3AA could be a new and promising antimicrobial agent for the combat of P. aeruginosa.