Interdependency of Respiratory Metabolism and Phenazine-Associated Physiology in Pseudomonas aeruginosa PA14

Jo, Jeanyoung; Price-Whelan, Alexa; Cornell, William Cole; Dietrich, Lars E. P.

doi:10.1128/jb.00700-19

Cited by 36 publications

(49 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The respiratory chain of P. aeruginosa includes several dehydrogenases, the cytochrome bc 1 complex, and five terminal oxidases that operate at different concentrations of oxygen, as well as alternative electron acceptors that operate under anaerobic conditions [9]. It has been shown that this bacterium selectively expresses different respiratory enzymes/pathways depending on the availability of nutrients, oxygen, and other electron acceptors [10][11][12][13][14]. Such adaptations are important for colonization of infection sites, particularly in the lungs of patients suffering CF, where the bacteria are challenged with low nutrient and oxygen availability [5].…”

Section: Introductionmentioning

confidence: 99%

The three NADH dehydrogenases of Pseudomonas aeruginosa: Their roles in energy metabolism and links to virulence

et al. 2021

View full text Add to dashboard Cite

Pseudomonas aeruginosa is a ubiquitous opportunistic pathogen which relies on a highly adaptable metabolism to achieve broad pathogenesis. In one example of this flexibility, to catalyze the NADH:quinone oxidoreductase step of the respiratory chain, P. aeruginosa has three different enzymes: NUO, NQR and NDH2, all of which carry out the same redox function but have different energy conservation and ion transport properties. In order to better understand the roles of these enzymes, we constructed two series of mutants: (i) three single deletion mutants, each of which lacks one NADH dehydrogenase and (ii) three double deletion mutants, each of which retains only one of the three enzymes. All of the mutants grew approximately as well as wild type, when tested in rich and minimal medium and in a range of pH and [Na+] conditions, except that the strain with only NUO (ΔnqrFΔndh) has an extended lag phase. During exponential phase, the NADH dehydrogenases contribute to total wild-type activity in the following order: NQR > NDH2 > NUO. Some mutants, including the strain without NQR (ΔnqrF) had increased biofilm formation, pyocyanin production, and killed more efficiently in both macrophage and mouse infection models. Consistent with this, ΔnqrF showed increased transcription of genes involved in pyocyanin production.

show abstract

Section: Introductionmentioning

confidence: 99%

The three NADH dehydrogenases of Pseudomonas aeruginosa: Their roles in energy metabolism and links to virulence

et al. 2021

View full text Add to dashboard Cite

show abstract

“…It is therefore possible that, in the oxic zone at 12 h, cells in SCFM are more protected from PYO toxicity than in LB at 24 h due to a higher electron donor:electron acceptor ratio (41). Finally, the ratios and production of different phenazines (e.g., PYO, PCA, PCN) in P. aeruginosa are known to vary depending on the carbon source in the growth medium (43); such variation could also contribute to the differences that we observed in PodA effects between our media.…”

Section: Discussionmentioning

confidence: 99%

Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms

VanDrisse

Lipsh‐Sokolik

Khersonsky

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

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.

show abstract

“…These approaches can allow cells to produce electrical current or consume it, resulting in either oxidation or reduction of intracellular redox species, respectively. Bioelectrochemical devices can then be used to drive biosynthetic reactions 1,4,5 , perform bioelectronic sensing 9 , actuate gene expression 3 , and modulate cellular growth 10,11 within the microorganism of interest. Despite these accomplishments, these strategies couple the redox state of the electrode to multiple intracellular redox biomolecules, resulting in off-target effects, cellular toxicity, and poor control of biosynthesis 1,3,4 .…”

Section: Figurementioning

confidence: 99%

Precise electronic control of redox reactions insideEscherichia coliusing a genetic module

Baruch

Tejedor-Sanz²,

Su³

et al. 2020

Preprint

View full text Add to dashboard Cite

18Microorganisms regulate the redox state of different biomolecules to precisely control 19 biological processes. These processes can be modulated by electrochemically coupling 20 intracellular biomolecules to an external electrode, but current approaches afford only 21 limited control and specificity. Here we describe specific electrochemical control of the 22 Microorganisms accomplish important biological functions such as conserving energy, 32 regulating gene expression, and powering biosynthesis using different redox-active 33 biomolecules. To enable control of these processes in any microorganism, researchers have 34 coupled the redox state of these biomolecules to an external electrode using membrane-35 permeable, small molecule redox mediators 1-5 , redox polymers 6 , and membrane-36 intercalated nanostructures 7,8 . These approaches can allow cells to produce electrical 37 current or consume it, resulting in either oxidation or reduction of intracellular redox species, 38 respectively. Bioelectrochemical devices can then be used to drive biosynthetic reactions 39 1,4,5 , perform bioelectronic sensing 9 , actuate gene expression 3 , and modulate cellular growth 40 10,11 within the microorganism of interest. Despite these accomplishments, these strategies 41 Results 75E. coli consume current using mtr and native oxidoreductases 76 We first sought to determine if the Mtr pathway could allow cathodic electrons to 77 directly enter a heterologous host upon addition of an electron acceptor. Since E. coli has 78 two MK-linked fumarate reductases, FrdABCD and SdhABCD, we hypothesized that the Mtr 79 pathway in E. coli could deliver cathodic electrons via these native proteins to fumarate 80 ( Figure 1A). To probe the specific role of the Mtr pathway, we compared the behavior of 81 several strains: E. coli expressing only the cytochrome c maturation (ccm) genes (abbrev. 82Ccm-E.coli) 17 , E. coli expressing ccm and mtrCAB (abbrev. Mtr-E. coli) 15 , and E. coli 83 expressing ccm and cymAmtrCAB (abbrev. CymAMtr-E. coli) 17 . The ccm genes are required 84 to make cytochromes c in the C43(DE3) parental background. 85To prepare E. coli for cathodic conditions, individual strains were first grown 86 aerobically, incubated in potentiostatic-controlled bioreactors under anaerobic, anodic 87 conditions (∆V=+200 mV Ag/AgCl ) for at least one day. Under these conditions, the CymAMtr-88 E. coli strain produced a significant steady-state current, while Ccm-E. coli and Mtr-E. coli 89 produced much lower currents (Figure 2A), reinforcing that CymA is important for current 90 production [16][17][18] . As anaerobic conditions were maintained, the electrode bias was then 91 switched to cathodic conditions (∆V= -560 V Ag/AgCl ), fumarate was added, and current 92 consumption was measured. In the absence of E. coli, neither current consumption nor 93 fumarate reduction was observed (Figure S1A). Likewise, the Ccm-E. coli strain did not 94 consume significant levels of current (Figure 2B). In contrast, both the Mtr-E. coli and t...

show abstract

Interdependency of Respiratory Metabolism and Phenazine-Associated Physiology in Pseudomonas aeruginosa PA14

Cited by 36 publications

References 60 publications

The three NADH dehydrogenases of Pseudomonas aeruginosa: Their roles in energy metabolism and links to virulence

The three NADH dehydrogenases of Pseudomonas aeruginosa: Their roles in energy metabolism and links to virulence

Computationally designed pyocyanin demethylase acts synergistically with tobramycin to kill recalcitrant Pseudomonas aeruginosa biofilms

Precise electronic control of redox reactions insideEscherichia coliusing a genetic module

Contact Info

Product

Resources

About