Andrew K. Marcus

A kinetic perspective on extracellular electron transfer by anode-respiring bacteria.

In microbial fuel cells and electrolysis cells (MXCs), anode-respiring bacteria (ARB) oxidize organic substrates to produce electrical current. In order to develop an electrical current, ARB must transfer electrons to a solid anode through extracellular electron transfer (EET). ARB use various EET mechanisms to transfer electrons to the anode, including direct contact through outer-membrane proteins, diffusion of soluble electron shuttles, and electron transport through solid components of the extracellular biofilm matrix. In this review, we perform a novel kinetic analysis of each EET mechanism by analyzing the results available in the literature. Our goal is to evaluate how well each EET mechanism can produce a high current density (> 10 A m(-2)) without a large anode potential loss (less than a few hundred millivolts), which are feasibility goals of MXCs. Direct contact of ARB to the anode cannot achieve high current densities due to the limited number of cells that can come in direct contact with the anode. Slow diffusive flux of electron shuttles at commonly observed concentrations limits current generation and results in high potential losses, as has been observed experimentally. Only electron transport through a solid conductive matrix can explain observations of high current densities and low anode potential losses. Thus, a study of the biological components that create a solid conductive matrix is of critical importance for understanding the function of ARB.

Proton transport inside the biofilm limits electrical current generation by anode‐respiring bacteria

Anode‐respiring bacteria (ARB) in a biofilm anode carry out an oxidation half‐reaction of organic matter, producing an electrical current from renewable biomass, including wastes. At the same time, ARB produce protons, usually one proton for every electron. Our study shows how current density generated by an acclimated ARB biofilm was limited by proton transport out of the biofilm. We determined that, at high current densities, protons were mainly transported out of the biofilm by protonating the conjugate base of the buffer system; the maximum current generation was directly related to the transport of the buffer, mainly by diffusion, into and out of the biofilm. With non‐limiting acetate concentrations, the current density increased with higher buffer concentrations, going from 2.21 ± 0.02 A m−2 with 12.5‐mM phosphate buffer medium to 9.3 ± 0.4 A m−2 using a 100‐mM phosphate buffer at a constant anode potential of Eanode = −0.35 V versus Ag/AgCl. Increasing the concentration of sodium chloride in the medium (0–100 mM) increased current density by only 15%, indicating that ion migration was not as important as diffusion of phosphate inside the biofilm. The current density also varied strongly with medium pH as a result of the buffer speciation: The current density was 10.0 ± 0.8 A m−2 at pH 8, and the pH giving one‐half the maximum rate was 6.5. A j–V curve analysis using 100 mM phosphate buffer showed a maximum current density of 11.5 ± 0.9 A m−2 and half‐saturation potential of −0.414 V versus Ag/AgCl, a value that deviated only slightly from the standard acetate potential, resulting in small anode‐potential losses. We discuss the implications of the proton‐transport limitation in the field of microbial fuel cells and microbial electrolytic cells. Biotechnol. Bioeng. 2008;100: 872–881.

Selecting Anode-Respiring Bacteria Based on Anode Potential: Phylogenetic, Electrochemical, and Microscopic Characterization

Anode-respiring bacteria (ARB) are able to transfer electrons contained in organic substrates to a solid electrode. The selection of ARB should depend on the anode potential, which determines the amount of energy available for bacterial growth and maintenance. In our study, we investigated how anode potential affected the microbial diversity of the biofilm community. We used a microbial electrolysis cell (MEC) containing four graphite electrodes, each at a different anode potential (E(anode) = -0.15, -0.09, +0.02, and +0.37 V vs SHE). We used wastewater-activated sludge as inoculum, acetate as substrate, and continuous-flow operation. The two electrodes at the lowest potentials showed a faster biofilm growth and produced the highest current densities, reaching up to 10.3 A/m(2) at the saturation of an amperometric curve; the electrode at the highest potential produced a maximum of 0.6 A/m(2). At low anode potentials, clone libraries showed a strong selection (92-99% of total clones) of an ARB that is 97% similar to G. sulfurreducens. At the highest anode potential, the ARB community was diverse. Cyclic voltammograms performed on each electrode suggest that the ARB grown at the lowest potentials carried out extracellular electron transport exclusively by conducting electrons through the extracellular biofilm matrix. This is supported by scanning electron micrographs showing putative bacterial nanowires and copious EPS at the lowest potentials. Non-ARB and ARB using electron shuttles in the diverse community for the highest anode potential may have insulated the ARB using a solid conductive matrix from the anode. Continuous-flow operation and the selective pressure due to low anode potentials selected for G. sulfurreducens, which are known to consume acetate efficiently and use a solid conductive matrix for electron transport.

Morphological synergism in root hair length, density, initiation and geometry for phosphorus acquisition in Arabidopsis thaliana: A modeling approach

Low phosphorus availability regulates root hair growth in Arabidopsis by (1) increasing root hair length, (2) increasing root hair density, (3) decreasing the distance between the root tip and the point at which root hairs begin to emerge, and (4) increasing the number of epidermal cell files that bear hairs (trichoblasts). The coordinated regulation of these traits by phosphorus availability prompted us to speculate that they are synergistic, that is, that they have greater adaptive value in combination than they do in isolation. In this study, we explored this concept using a geometric model to evaluate the effect of varying root hair length (short, medium, and long), density (0, 24, 48, 72, 96, and 120 root hairs per mm of root length), tip to first root hair distance (0.5, 1, 2, and 4 mm), and number of trichoblast files (8 vs. 12) on phosphorus acquisition efficiency (PAE) in Arabidopsis. SimRoot, a dynamic three-dimensional geometric model of root growth and architecture, was used to simulate the growth of Arabidopsis roots with contrasting root hair parameters at three values of phosphorus diffusion coefficient (De=1×10−7, 1×10−8, and 1×10−9 cm2 s−1) over time (20, 40, and 60 h). Depzone, a program that dynamically models nutrient diffusion to roots, was employed to estimate PAE and competition among root hairs. As De decreased from 1×10−7 to 1×10−9 cm2 s−1, roots with longer root hairs and higher root hair densities had greater PAE than those with shorter and less dense root hairs. At De=1×10−9 cm2 s−1, the PAE of root hairs at any given density was in the order of long hairs > medium length hairs > short hairs, and the maximum PAE occurred at density = 96 hairs mm−1 for both long and medium length hairs. This was due to greater competition among root hairs when they were short and dense. Competition over time decreased differences in PAE due to density, but the effect of length was maintained, as there was less competition among long hairs than short hairs. At high De(1×10−7 cm2 s−1), competition among root hairs was greatest among long hairs and lowest among short hairs, and competition increased with increasing root hair densities. This led to a decrease in PAE as root hair length and density increased. PAE was also affected by the tip to first root hair distance. At low De values, decreasing tip to first root hair distance increased PAE of long hairs more than that of short hairs, whereas at high De values, decreasing tip to first root hair distance increased PAE of root hairs at low density but decreased PAE of long hairs at very high density. Our models confirmed the benefits of increasing root hair density by increasing the number of trichoblast files rather than decreasing the trichoblast length. The combined effects of all four root hair traits on phosphorus acquisition was 371% greater than their additive effects, demonstrating substantial morphological synergy. In conclusion, our data support the hypothesis that the responses of root hairs to low phosphorus availability are synergistic, which may account for their coordinated regulation.

Analysis of a microbial electrochemical cell using the proton condition in biofilm (PCBIOFILM) model.

Common to all microbial electrochemical cells (MXCs) are the anode-respiring bacteria (ARB), which transfer electrons to an anode and release protons that must transport out of the biofilm. Here, we develop a novel modeling platform, Proton Condition in BIOFILM (PCBIOFILM), with a structure geared towards mechanistically explaining: (1) how the ARB half reaction produces enough acid to inhibit the ARB by low pH; (2) how the diffusion of alkalinity carriers (phosphates and carbonates) control the pH gradients in the biofilm anode; (3) how increasing alkalinity attenuates pH gradients and increases current; and (4) why carbonates enable higher current density than phosphates. Analysis of literature data using PCBIOFILM supports the hypothesis that alkalinity limits the maximum current density for MXCs. An alkalinity criterion for eliminating low-pH limitation - 12 mgCaCO(3)/mg BOD--implies that a practical MXC can achieve a maximum current density with an effluent quality comparable to anaerobic digestion.

Advanced control for photoautotrophic growth and CO2-utilization efficiency using a membrane carbonation photobioreactor (MCPBR).

A membrane carbonation (MC) module uses bubbleless gas-transfer membranes to supply inorganic carbon (C(i)) for photoautotrophic cyanobacterial growth in a photobioreactor (PBR); this creates the novel MCPBR system, which allows precise control of the CO(2)-delivery rate and minimal loss of CO(2) to the atmosphere. Experiments controlled the supply rate of C(i) to the main PBR by regulating the recirculation rate (Q(R)) between the module of MC chamber and the main PBR. The experiments evaluated how Q(R) controls the CO(2) mass transport in MC chamber and how it connects with the biomass production rate, C(i) concentration, pH in the PBR, and CO(2)-utilization efficiency. The biomass production rate and C(i) concentration increased in response to the C(i) supply rate (controlled by Q(R)), but not in linear proportion. The biomass production rate increased less than C(i) due to increased light limitation. Except for the highest Q(R), when the higher C(i) concentration caused the pH to decrease, CO(2) loss to gas ventilation was negligible. The results demonstrate that this MCPBR offers independent control over the growth of photoautotrophic biomass, pH control, and minimal loss of CO(2) to the atmosphere.

A Steady-State Biofilm Model for Simultaneous Reduction of Nitrate and Perchlorate, Part 2: Parameter Optimization and Results and Discussion

Part 1 of this work developed a steady-state, multispecies biofilm model for simultaneous reduction of nitrate and perchlorate in the H(2)-based membrane biofilm reactor (MBfR) and presented a novel method to solve it. In Part 2, the half-maximum-rate concentrations and inhibition coefficients of nitrate and perchlorate are optimized by fitting data from experiments with different combinations of influent nitrate and perchlorate concentrations. The model with optimized parameters is used to quantitatively and systematically explain how three important operating conditions (nitrate loading, perchlorate loading, and H(2) pressure) affect nitrate and perchlorate reduction and biomass distribution in these reducing biofilms. Perchlorate reduction and accumulation of perchlorate-reducing bacteria (PRB) in the biofilm are affected by four promotion or inhibition mechanisms: simultaneous use of nitrate and perchlorate by PRB and competition for H(2), the same resources in PRB, and space in a biofilm. For the hydrogen pressure evaluated experimentally, a low nitrate loading (<0.1 g N/m(2)-d) slightly promotes perchlorate removal, because of the beneficial effect from PRB using both acceptors. However, a nitrate loading of >0.6 g N/m(2)-d begins to inhibit perchlorate removal, as the competition effects become dominant.

A steady-state biofilm model for simultaneous reduction of nitrate and perchlorate, part 1: model development and numerical solution.

A multispecies biofilm model is developed for simultaneous reduction of nitrate and perchlorate in the H(2)-based membrane biofilm reactor. The one-dimension model includes dual-substrate Monod kinetics for a steady-state biofilm with five solid and five dissolved components. The solid components are autotrophic denitrifying bacteria, autotrophic perchlorate-reducing bacteria, heterotrophic bacteria, inert biomass, and extracellular polymeric substances (EPS). The dissolved components are nitrate, perchlorate, hydrogen (H(2)), substrate-utilization-associated products, and biomass-associated products (BAP). The model explicitly considers four mechanisms involved in how three important operating conditions (H(2) pressure, nitrate loading, and perchlorate loading) affect nitrate and perchlorate removals: (1) competition for H(2), (2) promotion of PRB growth due to having two electron acceptors (nitrate and perchlorate), (3) competition between nitrate and perchlorate reduction for the same resources in the PRB: electrons and possibly reductase enzymes, and (4) competition for space in the biofilm. Two other special features are having H(2) delivered from the membrane substratum and solving directly for steady state using a novel three-step approach: finite-difference for approximating partial differential and/or integral equations, Newton-Raphson for solving nonlinear equations, and an iterative scheme to obtain the steady-state biofilm thickness. An example result illustrates the models features.

Understanding the Distinguishing Features of a Microbial Fuel Cell as a Biomass-Based Renewable Energy Technology

Biomass-based renewable energy, which utilizes biomass derived from photosynthesis, could sustainably provide 67–450 EJ of energy annually. Biomass in organic wastes, for example, can annually provide 7.5 EJ of energy, and utilization of organic wastes locally as an energy source can prevent environmental pollution and reduce the energy losses associated with transportation. The technological challenge is to sustainably capture this biomass energy without creating serious environmental or social damage.

Performance of plastic- and sponge-based trickling filters treating effluents from an UASB reactor

The paper compares the performance of two trickling filters (TFs) filled with plastic- or sponge-based packing media treating the effluent from an upflow anaerobic sludge blanket (UASB) reactor. The UASB reactor was operated with an organic loading rate (OLR) of 1.2 kgCOD m(-3) d(-1), and the OLR applied to the TFs was 0.30-0.65 kgCOD m(-3) d(-1) (COD: chemical oxygen demand). The sponge-based packing medium (Rotosponge) gave substantially better performance for ammonia, total-N, and organic matter removal. The superior TF-Rotosponge performance for NH(4)(+)-N removal (80-95%) can be attributed to its longer biomass and hydraulic retention times (SRT and HRT), as well as enhancements in oxygen mass transfer by dispersion and advection inside the sponges. Nitrogen removals were significant (15 mgN L(-1)) in TF-Rotosponge when the OLRs were close to 0.75 kgCOD m(-3) d(-1), due to denitrification that was related to solids hydrolysis in the sponge interstices. For biochemical oxygen demand removal, higher HRT and SRT were especially important because the UASB removed most of the readily biodegradable organic matter. The new configuration of the sponge-based packing medium called Rotosponge can enhance the feasibility of scaling-up the UASB/TF treatment, including when retrofitting is necessary.