Characterization of nutrient removal and microalgal biomass production on an industrial waste-stream by application of the deceleration-stat technique.

Industrial wastewaters can serve as a nutrient and water source for microalgal production. In this study the effluent of an internal circulation (IC) reactor anaerobically treating the wastes of a biotechnology production facility were chosen as the cultivation medium for Chlorella sorokiniana in batch and continuous cultures. The aim was to evaluate the rates of nutrient removal and biomass production possible at various dilution rates. The results demonstrate that the industrial wastewater served as a highly effective microalgae culture medium and that dilution rate strongly influenced algae productivity in a short light-path photobioreactor. Batch culture on undiluted wastewater showed biomass productivity of 1.33 g L(-1)day(-1), while removing over 99% of the ammonia and phosphate from the wastewater. Deceleration-stat (D-stat) experiments performed at high and low intensities of 2100 and 200 (µmol photon m(2)s(-1)) established the optimal dilution rates to reach volumetric productivity of 5.87 and 1.67 g L(-1)day(-1) respectively. The corresponding removal rates of nitrogen were 238 and 93 mg L(-1)day(-1) and 40 and 19 mg L(-1)day(-1) for phosphorous. The yield on photons at low light intensity was as high as had been observed in any previous report indicating that the waste stream allowed the algae to grow at its full potential.

Microplate-based method for high-throughput screening of microalgae growth potential.

Microalgae cultivation conditions in microplates will differ from large-scale photobioreactors in crucial parameters such as light profile, mixing and gas transfer. Hence volumetric productivity (P(v)) measurements made in microplates cannot be directly scaled up. Here we demonstrate that it is possible to use microplates to measure characteristic exponential growth rates and determine the specific growth rate light intensity dependency (µ-I curve), which is useful as the key input for several models that predict P(v). Nannochloropsis salina and Chlorella sorokiniana specific growth rates were measured by repeated batch culture in microplates supplied with continuous light at different intensities. Exponential growth unlimited by gas transfer or self-shading was observable for a period of several days using fluorescence, which is an order of magnitude more sensitive than optical density. The microplate datasets were comparable to similar datasets obtained in photobioreactors and were used an input for the Huesemann model to accurately predict P(v).

Metabolic adaptations in a H2 producing heterocyst-forming cyanobacterium: potentials and implications for biological engineering.

Nostoc punctiforme ATCC 29133 is a photoautotrophic cyanobacterium with the ability to fix atmospheric nitrogen and photoproduce hydrogen through the enzyme nitrogenase. The H(2) produced is reoxidized by an uptake hydrogenase. Inactivation of the uptake hydrogenase in N. punctiforme leads to increased H(2) release but unchanged rates of N(2) fixation, indicating redirected metabolism. System-wide understanding of the mechanisms of this metabolic redirection was obtained using complementary quantitative proteomic approaches, at both the filament and the heterocyst level. Of the total 1070 identified and quantified proteins, 239 were differentially expressed in the uptake hydrogenase mutant (NHM5) as compared to wild type. Our results indicate that the inactivation of uptake hydrogenase in N. punctiforme changes the overall metabolic equilibrium, affecting both oxygen reduction mechanisms in heterocysts as well as processes providing reducing equivalents for metabolic functions such as N(2) fixation. We identify specific metabolic processes used by NHM5 to maintain a high rate of N(2) fixation, and thereby potential targets for further improvement of nitrogenase based H(2) photogeneration. These targets include, but are not limited to, components of the oxygen scavenging capacity and cell envelope of heterocysts and proteins directly or indirectly involved in reduced carbon transport from vegetative cells to heterocysts.