Ozonized biochar filtrate effects on the growth of Pseudomonas putida and cyanobacteria Synechococcus elongatus PCC 7942

Background Biochar ozonization was previously shown to dramatically increase its cation exchange capacity, thus improving its nutrient retention capacity. The potential soil application of ozonized biochar warrants the need for a toxicity study that investigates its effects on microorganisms. Results In the study presented here, we found that the filtrates collected from ozonized pine 400 biochar and ozonized rogue biochar did not have any inhibitory effects on the soil environmental bacteria Pseudomonas putida, even at high dissolved organic carbon (DOC) concentrations of 300 ppm. However, the growth of Synechococcus elongatus PCC 7942 was inhibited by the ozonized biochar filtrates at DOC concentrations greater than 75 ppm. Further tests showed the presence of some potential inhibitory compounds (terephthalic acid and p-toluic acid) in the filtrate of non-ozonized pine 400 biochar; these compounds were greatly reduced upon wet-ozonization of the biochar material. Nutrient detection tests also showed that dry-ozonization of rogue biochar enhanced the availability of nitrate and phosphate in its filtrate, a property that may be desirable for soil application. Conclusion Ozonized biochar substances can support soil environmental bacterium Pseudomonas putida growth, since ozonization detoxifies the potential inhibitory aromatic molecules. Graphical Abstract

Wastewater Substrate Disinfection for Application of Synechococcus Elongatus PCC 7942 Cultivation as Tertiary Treatment

The cultivation of microalgae or/and cyanobacteria in nutrient-rich wastewaters presents a significant opportunity for enhancing sustainability of tertiary wastewater treatment processes via resources/energy recovery/production. However, maintaining a monoculture in wastewater-media constitutes a significant challenge to be addressed, as a plethora of antagonistic and predating microorganisms exist is such media. In this regard, the present work assesses the efficiency of the low-cost wastewater substrate disinfection techniques of filtration, use of NaClO, H2O2 or Fe(VI), in terms of antagonistic or/and predating microbial species growth inhibition in Synechococcus elongatus PCC 7942 cultivations. Nitrates and phosphates removal rates were also experimentally assessed. The results showed that filter thickness has a greater effect on disinfection efficiency than that of filter’s pore size. Furthermore, the disinfection efficiency of Fe(VI), which was produced on-site by electrosynthesis via a Fe0/Fe0 cell, was greater than that of NaClO and H2O2. Filtration at ≤ 1.2 µm pore size coupled with chemical disinfection leads to unhindered S7942 growth and efficient nitrates and phosphates removal rates, at dosages of CT ≥ 270 mg min L−1 for NaClO and CT ≥ 157 mg min L−1 for Fe(VI). The coagulation action of Fe(III) species that result from Fe(VI) reduction and the oxidation action of Fe(VI) can assist in turbidity, organic compounds and phosphorous removal from wastewater-media. Moreover, the residual iron species can assist in S7942 harvesting and may enhance photosynthesis rate. Thus, the utilization of wastewaters for S7942 cultivation as tertiary treatment seems a promising and novel alternative to common nutrient removal processes that can reduce environmental footprint and operational costs of wastewater treatment plants.

Overexpression of Cu/Zn Superoxide Dismutase (Cu/Zn SOD) in Synechococcus elongatus PCC 7942 for Enhanced Azo Dye Removal through Hydrogen Peroxide Accumulation

Discharge of recalcitrant azo dyes to the environment poses a serious threat to environmental health. However certain microorganisms in nature have developed their survival strategies by degrading these toxic dyes. Cyanobacteria are one such prokaryotic, photosynthetic group of microorganisms that degrade various xenobiotic compounds, due to their capability to produce various reactive oxygen species (ROS), and particularly the hydrogen peroxide (H2O2) when released in their milieu. The accumulation of H2O2 is the result of the dismutation of superoxide radicals by the enzyme superoxide dismutase (SOD). In this study, we have genetically modified the cyanobacterium Synechococcus elongatus PCC 7942 by integrating Cu/Zn SOD gene (sodC) from Synechococcus sp. PCC 9311 to its neutral site through homologous recombination. The overexpression of sodC in the derivative strain was driven using a strong constitutive promoter of the psbA gene. The derivative strain resulted in constitutive production of sodC, which was induced further during dye-treated growth. The genetically engineered Synechococcus elongatus PCC 7942 (MS-sodC+) over-accumulated H2O2 during azo dye treatment with a higher dye removal rate than the wild-type strain (WS-sodC−). Therefore, enhanced H2O2 accumulation through SODs overexpression in cyanobacteria may serve as a valuable bioremediation tool.

Multi-layer Regulation of Rubisco in Response to Altered Carbon Status in Synechococcus elongatus PCC 7942

Photosynthetic organisms possess a variety of mechanisms to achieve balance between absorbed light (source) and the capacity to metabolically utilize or dissipate this energy (sink). While regulatory processes that detect changes in metabolic status/balance are relatively well-studied in plants, analogous pathways remain poorly characterized in photosynthetic microbes. Herein, we explore systemic changes that result from alterations in carbon availability in the model cyanobacterium Synechococcus elongatus PCC 7942 by taking advantage of an engineered strain where influx/efflux of a central carbon metabolite, sucrose, can be regulated experimentally. We observe that induction of a high-flux sucrose export pathway leads to depletion of internal carbon storage pools (glycogen), and concurrent increases in photosynthetic parameters. Further, a proteome-wide analysis and fluorescence reporter-based analysis revealed that upregulated factors following the activation of the metabolic sink are strongly concentrated on ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) and axillary modules involved in Rubisco maturation. Carboxysome number and Rubisco activity also increase following engagement of sucrose secretion. Conversely, reversing the flux of sucrose by feeding exogenous sucrose heterologously results in increased glycogen pools, decreased Rubisco abundance, decreased photosystem II quantum efficiency, and carboxysome reorganization. Our data suggest that Rubisco activity and organization are key outputs connected to regulatory pathways involved in metabolic balancing in cyanobacteria.

Shift in Conformational Equilibrium Underlies the Oscillatory Phosphoryl Transfer Reaction in the Circadian Clock

Oscillatory phosphorylation/dephosphorylation can be commonly found in a biological system as a means of signal transduction though its pivotal presence in the workings of circadian clocks has drawn significant interest: for example in a significant portion of the physiology of Synechococcus elongatus PCC 7942. The biological oscillatory reaction in the cyanobacterial circadian clock can be visualized through its reconstitution in a test tube by mixing three proteins—KaiA, KaiB and KaiC—with adenosine triphosphate and magnesium ions. Surprisingly, the oscillatory phosphorylation/dephosphorylation of the hexameric KaiC takes place spontaneously and almost indefinitely in a test tube as long as ATP is present. This autonomous post-translational modification is tightly regulated by the conformational change of the C-terminal peptide of KaiC called the “A-loop” between the exposed and the buried states, a process induced by the time-course binding events of KaiA and KaiB to KaiC. There are three putative hydrogen-bond forming residues of the A-loop that are important for stabilizing its buried conformation. Substituting the residues with alanine enabled us to observe KaiB’s role in dephosphorylating hyperphosphorylated KaiC, independent of KaiA’s effect. We found a novel role of KaiB that its binding to KaiC induces the A-loop toward its buried conformation, which in turn activates the autodephosphorylation of KaiC. In addition to its traditional role of sequestering KaiA, KaiB’s binding contributes to the robustness of cyclic KaiC phosphorylation by inhibiting it during the dephosphorylation phase, effectively shifting the equilibrium toward the correct phase of the clock.

Assessment of Synechococcus elongatus PCC 7942 as an option for sustainable wastewater treatment

Industrial wastewaters are recognized as a valuable resource, however their disposal without proper treatment can result in environmental deterioration. The associated environmental/operational cost of wastewater treatment necessitates upgrade of applied processes towards the goals of sustainability and mitigation of climate change. The implementation of cyanobacteria-based processes can contribute to these goals via resources recovery, production of high-value products, carbon fixation and green-energy production. The present study evaluates the cyanobacterium Synechococcus elongatus PCC 7942 (S7942) as a biological component for novel and sustainable alternatives to typical biological nutrient removal processes. Valuable results regarding cultivation temperature boundaries, applied disinfection techniques and analytical methods, as well as regarding relations between parameters expressing S7942 biomass concentration are presented. The results show that at typical industrial wastewater temperatures, S7942 efficiently grew and removed nitrates from treated snack-industry's wastewater. Moreover, in cultures with treated and relatively saline dairy wastewater, its growth rate slightly decreased, but nevertheless nitrates removal rate remained efficiently high. A comparison between typical denitrification processes and the proposed nutrient removal process indicated that a S7942-based system may constitute an alternative or a supplementary to denitrification process. Thus, Synechococcus elongatus PCC 7942 proved to be a potent candidate towards sustainable industrial wastewater treatment applications.

Carboxysome Mispositioning Alters Growth, Morphology, and Rubisco Level of the Cyanobacterium Synechococcus elongatus PCC 7942

Photosynthetic cyanobacteria are responsible for almost half of global CO 2 fixation. Due to eutrophication, rising temperatures, and increasing atmospheric CO 2 concentrations, cyanobacteria have gained notoriety for their ability to form massive blooms in both freshwater and marine ecosystems across the globe.