Machine-assisted cultivation and analysis of biofilms

Hansen, Silla; Kabbeck, Tobias; Radtke, Carsten P.; Krause, Susanne; Krolitzki, Eva; Peschke, Theo; Gasmi, Jannis; Rabe, Kersten S.; Wagner, Michael; Horn, Harald; Hubbuch, Jürgen; Gescher, Johannes; Niemeyer, Christof M.

doi:10.1038/s41598-019-45414-6

Cited by 23 publications

(18 citation statements)

References 45 publications

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“…2,30 With regard to the exoelectrogenic bacteria investigated here, possible applications go beyond the currently intensively researched biosensor and fuel cell systems. For example, the cultivation of biofilms in microfluidic systems is making substantial progress 31 and this technological platform is also being used for research into supramolecular and dissipative material systems. [32][33][34] We therefore believe that future advancements of fluidically controllable DNA materials systems can make an important contribution to the development of novel biotechnological production systems.…”

Section: Discussionmentioning

confidence: 99%

Cultivation of Exoelectrogenic Bacteria in Conductive DNA Nanocomposite Hydrogels Yields a Programmable Biohybrid Materials System

Rehnlund

Klein

et al. 2019

Preprint

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The use of living microorganisms integrated within electrochemical devices is an expanding field of research, with applications in microbial fuel cells, microbial biosensors or bioreactors. We describe the use of porous nanocomposite materials prepared by DNA polymerization of carbon nanotubes (CNT) and silica nanoparticles (SiNP) for the construction of a programmable biohybrid system containing the exoelectrogenic bacterium Shewanella oneidensis. We initially demonstrate the electrical conductivity of the CNT-containing DNA composite by employment of chronopotentiometry, electrochemical impedance spectroscopy, and cyclic voltammetry. Cultivation of Shewanella oneidensis in these materials shows that the exoelectrogenic bacteria populate the matrix of the composite, while non-exoelectrogenic Escherichia coli remain on its surface. Moreover, the ability to use extracellular electron transfer pathways is positively correlated with number of cells within the conductive synthetic biofilm matrix. The Shewanella containing composite remains stable for several days. Programmability of this biohybrid material system is demonstrated by on-demand release and degradation induced by a short-term enzymatic stimulus. The perspectives of this approach for technical applications are being discussed.

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Section: Discussionmentioning

confidence: 99%

Cultivation of Exoelectrogenic Bacteria in Conductive DNA Nanocomposite Hydrogels Yields a Programmable Biohybrid Materials System

Rehnlund

Klein

et al. 2019

Preprint

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“…To quantify the average concentration of cells and riboflavin within a biofilm or the biofilm-matrix, respectively, S. oneidensis cells were grown in M4 minimal medium under oxic conditions using 20 mM lactate as electron donor. The cells were cultured in triplicate in a microfluidic flow-through system with straight polydimethylsiloxane (PDMS) channels and a total volume of 354 μL (53). The microfluidic systems were inoculated for 2 h using a culture with an OD 600 of 0.08 and a flow rate of 3 ml/h.…”

Section: Methodsmentioning

confidence: 99%

Extracellular riboflavin induces anaerobic biofilm formation inShewanella oneidensis

Edel¹,

Sturm

Sturm-Richter³

et al. 2021

Preprint

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Some microorganisms can respire with extracellular electron acceptors using an extended electron transport chain to the cell surface. These organisms apply flavin molecules as cofactors to facilitate one-electron transfer catalysed by the terminal reductases and as endogenous electron shuttles. In the model organism Shewanella oneidensis, riboflavin production and excretion triggers a specific biofilm formation response that is initiated at a specific threshold concentration, similar to canonical quorum sensing molecules. Riboflavin-mediated messaging is based on the overexpression of the gene encoding the putrescin decarboxylase speC which leads to posttranscriptional overproduction of proteins involved in biofilm formation. Using a model of growth-dependent riboflavin production under batch and biofilm growth conditions, the number of cells necessary to produce the threshold concentration per time was deduced. Furthermore, our results indicate that specific retention of riboflavin in the biofilm matrix leads to localized concentrations which by far exceed the necessary threshold value.

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“…To quantify the average concentration of cells and riboflavin within a biofilm or the biofilm-matrix, respectively, S. oneidensis cells were grown in M4 minimal medium under oxic conditions using 20 mM lactate as electron donor. The cells were cultured in triplicate in a microfluidic flow-through system with straight polydimethylsiloxane (PDMS) channels and a total volume of 354 µL [53]. The microfluidic systems were inoculated for 2 h using a culture with an OD 600 of 0.08 and a flow rate of 3 ml/h.…”

Section: Quantification Of the Biofilm Volume Via Optical Coherence Tomography (Oct)mentioning

confidence: 99%

Extracellular riboflavin induces anaerobic biofilm formation in Shewanella oneidensis

Edel

Sturm

Sturm-Richter

et al. 2021

Biotechnol Biofuels

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Background Some microorganisms can respire with extracellular electron acceptors using an extended electron transport chain to the cell surface. This process can be applied in bioelectrochemical systems in which the organisms produce an electrical current by respiring with an anode as electron acceptor. These organisms apply flavin molecules as cofactors to facilitate one-electron transfer catalyzed by the terminal reductases and in some cases as endogenous electron shuttles. Results In the model organism Shewanella oneidensis, riboflavin production and excretion trigger a specific biofilm formation response that is initiated at a specific threshold concentration, similar to canonical quorum-sensing molecules. Riboflavin-mediated messaging is based on the overexpression of the gene encoding the putrescine decarboxylase speC which leads to posttranscriptional overproduction of proteins involved in biofilm formation. Using a model of growth-dependent riboflavin production under batch and biofilm growth conditions, the number of cells necessary to produce the threshold concentration per time was deduced. Furthermore, our results indicate that specific retention of riboflavin in the biofilm matrix leads to localized concentrations, which by far exceed the necessary threshold value. Conclusion This study describes a new quorum-sensing mechanism in S. oneidensis. Biofilm formation of S. oneidensis is induced by low concentrations of riboflavin resulting in an upregulation of the ornithine-decarboxylase speC. The results can be applied for the development of strains catalyzing increased current densities in bioelectrochemical systems.

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Machine-assisted cultivation and analysis of biofilms

Cited by 23 publications

References 45 publications

Cultivation of Exoelectrogenic Bacteria in Conductive DNA Nanocomposite Hydrogels Yields a Programmable Biohybrid Materials System

Cultivation of Exoelectrogenic Bacteria in Conductive DNA Nanocomposite Hydrogels Yields a Programmable Biohybrid Materials System

Extracellular riboflavin induces anaerobic biofilm formation inShewanella oneidensis

Extracellular riboflavin induces anaerobic biofilm formation in Shewanella oneidensis

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