Installing extra bicarbonate transporters in the cyanobacterium Synechocystis sp. PCC6803 enhances biomass production.

As a means to improve carbon uptake in the cyanobacterium Synechocystis sp. strain PCC6803, we engineered strains to contain additional inducible copies of the endogenous bicarbonate transporter BicA, an essential component of the CO2-concentrating mechanism in cyanobacteria. When cultured under atmospheric CO2 pressure, the strain expressing extra BicA transporters (BicA(+) strain) grew almost twice as fast and accumulated almost twice as much biomass as the control strain. When enriched with 0.5% or 5% CO2, the BicA(+) strain grew slower than the control but still showed a superior biomass production. Introducing a point mutation in the large C-terminal cytosolic domain of the inserted BicA, at a site implicated in allosteric regulation of transport activity, resulted in a strain (BicA(+)(T485G) strain) that exhibited pronounced cell aggregation and failed to grow at 5% CO2. However, the bicarbonate uptake capacity of the induced BicA(+)(T485G) was twice higher than for the wild-type strain. Metabolic analyses, including phenotyping by synchrotron-radiation Fourier transform Infrared spectromicroscopy, scanning electron microscopy, and lectin staining, suggest that the excess assimilated carbon in BicA(+) and BicA(+)(T485G) cells was directed into production of saccharide-rich exopolymeric substances. We propose that the increased capacity for CO2 uptake in the BicA(+) strain can be capitalized on by re-directing carbon flux from exopolymeric substances to other end products such as fuels or high-value chemicals.

Geometrically asymmetric electrodes for probing electrochemical reaction kinetics: a case study of hydrogen at the Pt-CsH2PO4 interface.

Electrochemical reactions can exhibit considerable asymmetry, with the polarization behavior of oxidation at a given metal|electrolyte interface differing substantially from that of reduction. The reference-less, microcontact electrode geometry, in which the electrode overpotentials are geometrically constrained to the working electrode (by limiting its area) is experimentally convenient, particularly for fuel cell studies, because the results do not rely on accurate placement of a reference electrode nor must oxidant and reductant gases be sealed off from one another. Here, the conditions under which the critical assumption of this geometry applies-that the overpotential at the large-area counter electrode can be ignored-is numerically assessed. It is found that, for cells of sufficiently large area, the effective radius of the counter electrode (which defines the area through which the majority of the current passes) can be expressed directly as a function of electrolyte thickness and the materials properties, sigma, the conductivity of the electrolyte, and k, the reaction rate constant for the electrochemical reaction at zero-bias. From this effective radius and the true radius of the working electrode, the fraction of electrode overpotential at the latter, defined as the extent of isolation, can be readily computed. Experimental studies of hydrogen electro-oxidation/proton electro-reduction at the Pt|CsH(2)PO(4) interface using two cells of differing dimensions both validate the computational results and demonstrate that asymmetry in such reactions are readily revealed in the micro-electrode, reference-less geometry. The study furthermore confirms the insensitivity of the results to the precise placement of the working electrode, while indicating the importance of very high isolation values (>99%) to ensure that overpotential contributions of the counter electrode do not influence the measurements, particularly as bias is increased.