气候变化领域集成服务门户(CCPortal): Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms

CCPortal

DOI	10.1073/pnas.2022012118
	Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms
	VanDrisse C.M.; Lipsh-Sokolik R.; Khersonsky O.; Fleishman S.J.; Newman D.K.
发表日期	2021
ISSN	00278424
卷号	118 期号:12
英文摘要	Pseudomonas aeruginosa is an opportunistic human pathogen that develops difficult-to-treat biofilms in immunocompromised individuals, cystic fibrosis patients, and in chronic wounds. P. aeruginosa has an arsenal of physiological attributes that enable it to evade standard antibiotic treatments, particularly in the context of biofilms where it grows slowly and becomes tolerant to many drugs. One of its survival strategies involves the production of the redox-active phenazine, pyocyanin, which promotes biofilm development. We previously identified an enzyme, PodA, that demethylated pyocyanin and disrupted P. aeruginosa biofilm development in vitro. Here, we asked if this protein could be used as a potential therapeutic for P. aeruginosa infections together with tobramycin, an antibiotic typically used in the clinic. A major roadblock to answering this question was the poor yield and stability of wild-type PodA purified from standard Escherichia coli overexpression systems. We hypothesized that the insufficient yields were due to poor packing within PodA’s obligatory homotrimeric interfaces. We therefore applied the protein design algorithm, AffiLib, to optimize the symmetric core of this interface, resulting in a design that incorporated five mutations leading to a 20-fold increase in protein yield from heterologous expression and purification and a substantial increase in stability to environmental conditions. The addition of the designed PodA with tobramycin led to increased killing of P. aeruginosa cultures under oxic and hypoxic conditions in both the planktonic and biofilm states. This study highlights the potential for targeting extracellular metabolites to assist the control of P. aeruginosa biofilms that tolerate conventional antibiotic treatment. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Pseudomonas aeruginosa \| biofilms \| antibiotic tolerance \| pyocyanin \| AffiLib
语种	英语
scopus关键词	bacterial enzyme; enzyme PodA; pyocyanine; tobramycin; unclassified drug; algorithm; Article; bactericidal activity; bacterium culture; biofilm; controlled study; demethylation; drug tolerance; enzyme purification; enzyme stability; Escherichia coli; heterologous expression; hypoxia; mutation; nonhuman; plankton; priority journal; Pseudomonas aeruginosa; Pseudomonas infection; synergistic effect
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180177
作者单位	Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, United States; Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, 7610001, Israel; Division of Geological and Planetary Sciences, California Institute of Technology, Pasadena, CA 91125, United States
推荐引用方式 GB/T 7714	VanDrisse C.M.,Lipsh-Sokolik R.,Khersonsky O.,et al. Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms[J],2021,118(12).
APA	VanDrisse C.M.,Lipsh-Sokolik R.,Khersonsky O.,Fleishman S.J.,&Newman D.K..(2021).Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms.Proceedings of the National Academy of Sciences of the United States of America,118(12).
MLA	VanDrisse C.M.,et al."Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms".Proceedings of the National Academy of Sciences of the United States of America 118.12(2021).