Electrochemical Surface-Enhanced Raman Spectroscopy of Pyocyanin Secreted by Pseudomonas aeruginosa Communities

Do, Hyein; Kwon, Seung-Ryong; Fu, Kaiyu; Morales-Soto, Nydia; Shrout, Joshua D.; Bohn, Paul W.

doi:10.1021/acs.langmuir.9b00184

Electrochemical Surface-Enhanced Raman Spectroscopy of Pyocyanin Secreted by Pseudomonas aeruginosa Communities

Journal Article · Wed May 01 00:00:00 EDT 2019 · Langmuir

DOI:https://doi.org/10.1021/acs.langmuir.9b00184· OSTI ID:1765171

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Univ. of Notre Dame, IN (United States); University of Notre Dame
Univ. of Notre Dame, IN (United States)

Pyocyanin (PYO) is one of many toxins secreted by the opportunistic human pathogenic bacterium Pseudomonas aeruginosa. Direct detection of PYO in biofilms is crucial because PYO can provide important information about infection-related virulence mechanisms in P. aeruginosa. Because PYO is both redox-active and Raman-active, we seek to simultaneously acquire both spectroscopic and redox state information about PYO. The combination of surface-enhanced Raman spectroscopy (SERS) and voltammetry is used here to provide insights into the molecular redox behavior of PYO while controlling the SERS and electrochemical (EC) response of PYO with external stimuli, such as pH and applied potential. Furthermore, PYO secretion from biofilms of different P. aeruginosa strains is compared. Both SERS spectra and EC behavior are observed to change with pH, and several pH-dependent bands are identified in the SERS spectra, which can potentially be used to probe the local environment. Comparison of the voltammetric behavior of wild-type and a PYO-deficient mutant unequivocally identifies PYO as a major component of the secretome. Spectroelectrochemical studies of the PYO standard reveal decreasing SERS intensities of PYO bands under reducing conditions. Extending these experiments to pellicle biofilms shows similar behavior with applied potential, and SERS imaging indicates that secreted PYO is localized in regions approximately the size of P. aeruginosa cells. Furthermore, the in situ spectroelectrochemical biofilm characterization approach developed here suggests that EC–SERS monitoring of secreted molecules can be used diagnostically and correlated with the progress of infection.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Notre Dame, IN (United States)

Sponsoring Organization:: NIH; National Inst. of Allergy and Infectious Diseases (NIAID); USDOE Office of Science (SC)

Grant/Contract Number:: SC0019312

OSTI ID:: 1765171

Alternate ID(s):: OSTI ID: 1613171

Journal Information:: Langmuir, Journal Name: Langmuir Journal Issue: 21 Vol. 35; ISSN 0743-7463

Publisher:: American Chemical SocietyCopyright Statement

Country of Publication:: United States

Language:: English

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