Electrochemical camera chip for simultaneous imaging of multiple metabolites in biofilms

Bellin, Daniel L.; Sakhtah, Hassan; Zhang, Yihan; Price-Whelan, Alexa; Dietrich, Lars E. P.; Shepard, Kenneth L.

doi:10.1038/ncomms10535

Cited by 105 publications

(91 citation statements)

References 45 publications

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“…S9C), possibly due to degradation or modification during the prolonged agar extraction protocol. Thus, we sought to demonstrate a role for MexGHIOpmD in 5-Me-PCA release from colonies by using an integrated circuit (IC) chip, an electrochemical metabolite sensing platform recently developed by our group (48,54). This technique enables direct detection of phenazines as they are released from the biofilm using the same SWV-based electrochemical detection modality described earlier, eliminating the need for agar processing and extraction steps that may allow conversion of 5-Me-PCA to other derivatives before HPLC analysis (SI Appendix, Fig.…”

Section: Mexghi-opmd Contributes To Fitness In Shaken Liquid Culturesmentioning

confidence: 99%

The Pseudomonas aeruginosa efflux pump MexGHI-OpmD transports a natural phenazine that controls gene expression and biofilm development

Sakhtah

Koyama

Zhang

et al. 2016

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

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148

159

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Redox-cycling compounds, including endogenously produced phenazine antibiotics, induce expression of the efflux pump MexGHIOpmD in the opportunistic pathogen Pseudomonas aeruginosa. Previous studies of P. aeruginosa virulence, physiology, and biofilm development have focused on the blue phenazine pyocyanin and the yellow phenazine-1-carboxylic acid (PCA). In P. aeruginosa phenazine biosynthesis, conversion of PCA to pyocyanin is presumed to proceed through the intermediate 5-methylphenazine-1-carboxylate (5-Me-PCA), a reactive compound that has eluded detection in most laboratory samples. Here, we apply electrochemical methods to directly detect 5-Me-PCA and find that it is transported by MexGHIOpmD in P. aeruginosa strain PA14 planktonic and biofilm cells. We also show that 5-Me-PCA is sufficient to fully induce MexGHI-OpmD expression and that it is required for wild-type colony biofilm morphogenesis. These physiological effects are consistent with the high redox potential of 5-Me-PCA, which distinguishes it from other well-studied P. aeruginosa phenazines. Our observations highlight the importance of this compound, which was previously overlooked due to the challenges associated with its detection, in the context of P. aeruginosa gene expression and multicellular behavior. This study constitutes a unique demonstration of efflux-based selfresistance, controlled by a simple circuit, in a Gram-negative pathogen.phenazine | RND efflux | MexGHI-OpmD | biofilm | antibiotic

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Section: Mexghi-opmd Contributes To Fitness In Shaken Liquid Culturesmentioning

confidence: 99%

The Pseudomonas aeruginosa efflux pump MexGHI-OpmD transports a natural phenazine that controls gene expression and biofilm development

Sakhtah

Koyama

Zhang

et al. 2016

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

Self Cite

148

159

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“…PCA and other phenazines facilitate extracellular anaerobic respiration (Rabaey et al ., ; Wang et al ., ; Glasser et al ., ). Moreover, phenazine biosynthesis is regulated by quorum sensing (Pierson et al ., ; Khan et al ., ; O'Loughlin et al ., ) and oxygen (O 2 ) status (Bellin et al ., ). These findings indicate that phenazines play a role in community‐regulated respiration.…”

Section: Introductionmentioning

confidence: 99%

Phenazine‐1‐carboxylic acid and soil moisture influence biofilm development and turnover of rhizobacterial biomass on wheat root surfaces

LeTourneau

Marshall

Cliff

et al. 2018

Environmental Microbiology

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Phenazine-1-carboxylic acid (PCA) is produced by rhizobacteria in dryland but not in irrigated wheat fields of the Pacific Northwest, USA. PCA promotes biofilm development in bacterial cultures and bacterial colonization of wheat rhizospheres. However, its impact upon biofilm development has not been demonstrated in the rhizosphere, where biofilms influence terrestrial carbon and nitrogen cycles with ramifications for crop and soil health. Furthermore, the relationships between soil moisture and the rates of PCA biosynthesis and degradation have not been established. In this study, expression of PCA biosynthesis genes was upregulated relative to background transcription, and persistence of PCA was slightly decreased in dryland relative to irrigated wheat rhizospheres. Biofilms in dryland rhizospheres inoculated with the PCA-producing (PCA ) strain Pseudomonas synxantha 2-79RN were more robust than those in rhizospheres inoculated with an isogenic PCA-deficient (PCA ) mutant strain. This trend was reversed in irrigated rhizospheres. In dryland PCA rhizospheres, the turnover of N-labelled rhizobacterial biomass was slower than in the PCA and irrigated PCA treatments, and incorporation of bacterial N into root cell walls was observed in multiple treatments. These results indicate that PCA promotes biofilm development in dryland rhizospheres, and likely influences crop nutrition and soil health in dryland wheat fields.

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“…Although only pyocyanin is exclusively produced by PA, the simultaneous detection of several metabolites is advantageous both for bacterial research purposes, but also for using the presence of the compounds as a molecular signature for PA. Several studies have conducted multiple molecule detection using electrochemical measurements. These studies have mapped the potential values belonging to the aforementioned metabolites and do not overlap, which allows the simultaneous detection of the compounds in one measurement, e.g., through DPV or SWV in the relative potential range of −0.7 to 1.5 V [ 35 , 41 , 48 , 49 , 50 , 51 , 52 ].…”

Section: Electrochemical Detection Of Pyocyaninmentioning

confidence: 99%

“…Another study demonstrated the simultaneous electrochemical measuring and imaging of pyocyanin and other metabolites in biofilms using an SECM coupled to an electrochemical camera chip. This approach gives a unique resolution of the electrochemical activity of biofilms [ 50 ]. It is worth noticing that in the SECM setups, redox mediators have been used to generate a feedback approach curve.…”

Section: Electrochemical Detection Of Pyocyaninmentioning

confidence: 99%

Electrochemical Detection of Pyocyanin as a Biomarker for Pseudomonas aeruginosa: A Focused Review

Alatraktchi

Svendsen

Molin

2020

Sensors

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Pseudomonas aeruginosa (PA) is a pathogen that is recognized for its advanced antibiotic resistance and its association with serious diseases such as ventilator-associated pneumonia and cystic fibrosis. The ability to rapidly detect the presence of pathogenic bacteria in patient samples is crucial for the immediate eradication of the infection. Pyocyanin is one of PA’s virulence factors used to establish infections. Pyocyanin promotes virulence by interfering in numerous cellular functions in host cells due to its redox-activity. Fortunately, the redox-active nature of pyocyanin makes it ideal for detection with simple electrochemical techniques without sample pretreatment or sensor functionalization. The previous decade has seen an increased interest in the electrochemical detection of pyocyanin either as an indicator of the presence of PA in samples or as a tool for quantifying PA virulence. This review provides the first overview of the advances in electrochemical detection of pyocyanin and offers an input regarding the future directions in the field.

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Electrochemical camera chip for simultaneous imaging of multiple metabolites in biofilms

Cited by 105 publications

References 45 publications

The Pseudomonas aeruginosa efflux pump MexGHI-OpmD transports a natural phenazine that controls gene expression and biofilm development

The Pseudomonas aeruginosa efflux pump MexGHI-OpmD transports a natural phenazine that controls gene expression and biofilm development

Phenazine‐1‐carboxylic acid and soil moisture influence biofilm development and turnover of rhizobacterial biomass on wheat root surfaces

Electrochemical Detection of Pyocyanin as a Biomarker for Pseudomonas aeruginosa: A Focused Review

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