Interactive Effects of Ceftriaxone and Chitosan Immobilization on the Production of Arachidonic Acid by and the Microbiome of the Chlorophyte Lobosphaera sp. IPPAS C-2047.

Pharmaceuticals including antibiotics are among the hazardous micropollutants (HMP) of the environment. Incomplete degradation of the HMP leads to their persistence in water bodies causing a plethora of deleterious effects. Conventional wastewater treatment cannot remove HMP completely and a promising alternative comprises biotechnologies based on microalgae. The use of immobilized microalgae in environmental biotechnology is advantageous since immobilized cultures allow the recycling of the microalgal cells, support higher cell densities, and boost tolerance of microalgae to stresses including HMP. Here, we report on a comparative study of HMP (exemplified by the antibiotic ceftriaxone, CTA) removal by suspended and chitosan-immobilized cells of Lobosphaera sp. IPPAS C-2047 in flasks and in a column bioreactor. The removal of CTA added in the concentration of 20 mg/L was as high as 65% (in the flasks) or 85% (in the bioreactor). The adsorption on the carrier and abiotic oxidation were the main processes contributing 65-70% to the total CTA removal, while both suspended and immobilized cells took up 25-30% of CTA. Neither the immobilization nor CTA affected the accumulation of arachidonic acid (ARA) by Lobosphaera sp. during bioreactor tests but the subsequent nitrogen deprivation increased ARA accumulation 2.5 and 1.7 times in the suspended and chitosan-immobilized microalgae, respectively. The study of the Lobosphaera sp. microbiome revealed that the immobilization of chitosan rather than the CTA exposure was the main factor displacing the taxonomic composition of the microbiome. The possibility and limitations of the use of chitosan-immobilized Lobosphaera sp. IPPAS C-2047 for HMP removal coupled with the production of valuable long-chain polyunsaturated fatty acids is discussed.

Failure of Micractinium simplicissimum Phosphate Resilience upon Abrupt Re-Feeding of Its Phosphorus-Starved Cultures.

Microalgae are naturally adapted to the fluctuating availability of phosphorus (P) to opportunistically uptake large amounts of inorganic phosphate (Pi) and safely store it in the cell as polyphosphate. Hence, many microalgal species are remarkably resilient to high concentrations of external Pi. Here, we report on an exception from this pattern comprised by a failure of the high Pi-resilience in strain Micractinium simplicissimum IPPAS C-2056 normally coping with very high Pi concentrations. This phenomenon occurred after the abrupt re-supplementation of Pi to the M. simplicissimum culture pre-starved of P. This was the case even if Pi was re-supplemented in a concentration far below the level toxic to the P-sufficient culture. We hypothesize that this effect can be mediated by a rapid formation of the potentially toxic short-chain polyphosphate following the mass influx of Pi into the P-starved cell. A possible reason for this is that the preceding P starvation impairs the capacity of the cell to convert the newly absorbed Pi into a "safe" storage form of long-chain polyphosphate. We believe that the findings of this study can help to avoid sudden culture crashes, and they are also of potential significance for the development of algae-based technologies for the efficient bioremoval of P from P-rich waste streams.

The Escherichia coli Amino Acid Uptake Protein CycA: Regulation of Its Synthesis and Practical Application in l-Isoleucine Production.

Amino acid transport systems perform important physiological functions; their role should certainly be considered in microbial production of amino acids. Typically, in the context of metabolic engineering, efforts are focused on the search for and application of specific amino acid efflux pumps. However, in addition, importers can also be used to improve the industrial process as a whole. In this study, the protein CycA, which is known for uptake of nonpolar amino acids, was characterized from the viewpoint of regulating its expression and range of substrates. We prepared a cycA-overexpressing strain and found that it exhibited high sensitivity to branched-chain amino acids and their structural analogues, with relatively increased consumption of these amino acids, suggesting that they are imported by CycA. The expression of cycA was found to be dependent on the extracellular concentrations of substrate amino acids. The role of some transcription factors in cycA expression, including of Lrp and Crp, was studied using a reporter gene construct. Evidence for the direct binding of Crp to the cycA regulatory region was obtained using a gel-retardation assay. The enhanced import of named amino acids due to cycA overexpression in the l-isoleucine-producing strain resulted in a significant reduction in the generation of undesirable impurities. This work demonstrates the importance of uptake systems with respect to their application in metabolic engineering.

Sunscreen Effect Exerted by Secondary Carotenoids and Mycosporine-like Amino Acids in the Aeroterrestrial Chlorophyte Coelastrella rubescens under High Light and UV-A Irradiation.

The microalga Coelastrella rubescens dwells in habitats with excessive solar irradiation; consequently, it must accumulate diverse compounds to protect itself. We characterized the array of photoprotective compounds in C. rubescens. Toward this goal, we exposed the cells to high fluxes of visible light and UV-A and analyzed the ability of hydrophilic and hydrophobic extracts from the cells to absorb radiation. Potential light-screening compounds were profiled by thin layer chromatography and UPLC-MS. Coelastrella accumulated diverse carotenoids that absorbed visible light in the blue-green part of the spectrum and mycosporine-like amino acids (MAA) that absorbed the UV-A. It is the first report on the occurrence of MAA in Coelastrella. Two new MAA, named coelastrin A and coelastrin B, were identified. Transmission electron microscopy revealed the development of hydrophobic subcompartments under the high light and UV-A exposition. We also evaluate and discuss sporopollenin-like compounds in the cell wall and autophagy-like processes as the possible reason for the decrease in sunlight absorption by cells, in addition to inducible sunscreen accumulation. The results suggested that C. rubescens NAMSU R1 accumulates a broad range of valuable photoprotective compounds in response to UV-A and visible light irradiation, which indicates this strain as a potential producer for biotechnology.

All common bipedal gaits emerge from a single passive model.

In this paper, we systematically investigate passive gaits that emerge from the natural mechanical dynamics of a bipedal system. We use an energetically conservative model of a simple spring-leg biped that exhibits well-defined swing leg dynamics. Through a targeted continuation of periodic motions of this model, we systematically identify different gaits that emerge from simple bouncing in place. We show that these gaits arise along one-dimensional manifolds that bifurcate into different branches with distinctly different motions. The branching is associated with repeated breaks in symmetry of the motion. Among others, the resulting passive dynamic gaits include walking, running, hopping, skipping and galloping. Our work establishes that the most common bipedal gaits can be obtained as different oscillatory motions (or nonlinear modes) of a single mechanical system with a single set of parameter values. For each of these gaits, the timing of swing leg motion and vertical motion is matched. This work thus supports the notion that different gaits are primarily a manifestation of the underlying natural mechanical dynamics of a legged system. Our results might explain the prevalence of certain gaits in nature, and may provide a blueprint for the design and control of energetically economical legged robots.