Analytische Chemie
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Chemical and electrochemical interaction mechanisms of metal-reducing bacteria with gold surfaces
(2016)
Bacterial biofilms are considered one of the salient contributing factors to the deterioration of metals and their alloys, occurring in virtually all environments and across various industrial systems. Considering the sheer magnitude of detrimental effects, it is of pertinent interest to elucidate the interaction mechanisms of sessile bacteria with metal and metal oxide surfaces to facilitate the development of efficient antifouling strategies. A common constituent of microbial communities within aquatic and sedimentary settings, the Shewanella genus consists of facultatively aerobic, Gram-negative bacterium which exhibit exceptional plasticity in respiratory capacities. During aerobic conditions, Shewanella utilizes oxygen as a terminal electron acceptor; conversely, under anaerobic conditions, it is able to undertake respiration by reducing alternative terminal electron acceptors such as oxidized metals via extracellular electron transfer mechanisms not yet thoroughly discerned.
The aim of this work is to explicate the mechanisms governing the initial bacterial adhesion and subsequent biofilm formation on metallic surfaces. To investigate this dynamic interplay, a combined approach has been followed which couples surface enhanced Raman spectroscopy (SERS) with electrochemical techniques using Shewanella sp. model biofilms. Gold nano-islands deposited on thin glass slides have been chosen as inert model substrates with good uniformity and high surface enhancement factor. Furthermore, the utilization of gold as substrate material not only allowed the differentiation of the sole effect of substrate polarization on bacterial attachment but also enabled a precise adjustment of the surface chemistry and surface energy by means of surface functionalization with organothiol self-assembled monolayers.
The results present the correlation of the primary settlement rate of bacteria on metallic substrates with the environmental parameters such as electrolyte composition and pH as well as surface-related properties like hydrophobicity/hydrophilicity and polarization. With the overall strategic goal of transferring this methodology to technical systems the results provide the fundamental basis for the bottom-up design of anti-fouling surfaces.
Bacterial biofilms represent a ubiquitous form of microbial life on Earth. Due to an evolved armory of protean biological responses to external stimuli, bacteria are able to adhere to, colonize and thrive on virtually all surfaces, whether natural or synthetic, even in challenging environmental conditions. In addition to significant health risks, biofilms are among the salient contributors to the deterioration of metals and their alloys, thereby causing safety risks for technical equipment. Hence, understanding the interaction mechanisms of electroactive sessile bacteria with metal surfaces is vital for facilitating the development of efficient control strategies and novel anti-fouling surfaces in various industries and technologies.
The present study focusses on a combined spectroelectrochemical approach, melding methods of surface enhanced Raman spectroscopy (SERS) and electrochemical techniques, to investigate the chemical characteristics and redox activities of electroactive bacteria during the initial stages of biofilm formation. Gold has been selected as a model substrate due to its inert character, considerably high surface enhancement factor, as well as its capability to allow surface chemistry modifications and substrate polarization in order to precisely control the surface charge. Square wave voltammetry (SWV) and cyclic voltammetry (CV) studies have been performed for quantitative determination of flavin concentration and electrochemical impedance spectroscopy (EIS) has been utilized to study the changes in electrochemical processes within biofilms during different stages of growth. Shewanella sp. have been chosen as microorganisms within this work due to their versatile exoelectrogenic respiratory behavior and their distinct ability to reduce metals via extracellular electron transfer mechanisms involving self-secreted electron shuttle redox molecules such as flavins. To further explicate the process of diffusion of flavins within biofilms, a model system has been developed to simulate the structural features of the bacterial extracellular polymeric substances typically found in biofilms. This has been achieved by creating hydrogel films comprised of calcium-cross-linked alginate. The results demonstrate an interplay of factors contributing to the initial phases of bacterial settlement and biofilm formation as a function of environmental parameters. Furthermore, the results allow insight into the diffusion of flavins, much like they would in a natural biofilm, and how their redox behavior affects the biofilm development.
Bacterial biofilms have the capacity to develop and thrive in virtually all circumstances and surfaces, even in the most challenging environmental conditions. The pervasive and recalcitrant existence of biofilms renders them to be a significant safety risk and economical encumbrance in a wide array of industries and technologies, and therefore is a priority area of research.
It is essential to develop an improved understanding of the mechanisms implicated during biofilm formation, such as in the case of the diffusion of bacterial-secreted extracellular electron transporters, which are purported to play an important role during biocorrosion by exoelectrogenic bacteria. Hence, a fundamental understanding of electron transfer mechanisms between bacteria and extracellular electron acceptors will contribute insight to our understanding of charge transport and chemistry at the biofilm – external insoluble electron acceptor interface.
In the present work, Attenuated Total Reflection - Fourier transform-infrared (ATR-FTIR) spectroscopy has been coupled to electrochemical techniques for the nondestructive, in situ spectro-electrochemical monitoring of biofilms in real-time. Shewanella sp. have been selected for this investigation due to their adaptable exoelectrogenic respiratory capacities and their notable ability to reduce metals via several different mechanisms of extracellular electron transfer mechanisms, including self-secreted flavin shuttles. Gold-thin film model substrates have been used due to their inert nature and for their ability to permit precise manipulation of the substrate surface polarization. Additionally, hydrogels comprised of calcium cross-linked alginate have been used to mimic the architectural features of extracellular polymeric substances which are integral to a bacterial biofilm, to allow the study of electron-transporting flavin molecules in an artificial biofilm. The results will demonstrate patterns of diffusion, akin to how flavins would diffuse in a naturally occurring biofilm, and how polarization affects this process. Furthermore, insight will be gained on how the redox behavior of flavins can influence the development and evolution of a biofilm.
This interdisciplinary approach should shed light on bacterial electron transfer mechanisms which could contribute towards emerging technologies which seek to better understand such mechanisms for novel antifouling strategies, renewable energies, and bioremediation.