Cultivation of Chlorella sorokiniana IPPAS C-1 in Flat-Panel Photobioreactors: From a Laboratory to a Pilot Scale.

Flat-panel photobioreactors are effective systems for microalgae cultivation. This paper presents the growth characteristics of the microalgae Chlorella sorokiniana IPPAS C-1 as a result of three-stage scale-up cultivation in a specially designed cultivation system. First, C. sorokiniana was grown aseptically in 250 mL glass vessels; then, it was diluted and inoculated into a 5-liter flat-panel horizontal photobioreactor; and, at the last stage, the culture was diluted and inoculated into a 70-liter flat-panel vertical photobioreactor. In the presented cycle, the cultured biomass increased by 326 times in 13 days (from 0.6 to 195.6 g dw), with a final biomass concentration of 2.8 g dw L-1. The modes of semi-continuous cultivation were considered. The biomass harvest and dilution of the suspension were carried out either every day or every 3-4 days. For C. sorokiniana IPPAS C-1, a conversion coefficient of optical density values to dry biomass (g L-1) was refined through a factor of 0.33. The key parameters of the photobioreactors tested in this work are discussed.

Hydrogen Peroxide Participates in Perception and Transduction of Cold Stress Signal in Synechocystis.

The double mutant ΔkatG/tpx of cyanobacterium Synechocystis sp. strain PCC 6803, defective in the anti-oxidative enzymes catalase (KatG) and thioredoxin peroxidase (Tpx), is unable to grow in the presence of exogenous H2O2. The ΔkatG/tpx mutant is shown to be extremely sensitive to very low concentrations of H2O2, especially when intensified with cold stress. Analysis of gene expression in both wild-type and ΔkatG/tpx mutant cells treated by combined cold/oxidative stress revealed that H2O2 participates in regulation of expression of cold-responsive genes, affecting either signal perception or transduction. The central role of a transmembrane stress-sensing histidine kinase Hik33 in the cold/oxidative signal transduction pathway is discussed.

Putative extracellular α-class carbonic anhydrase, EcaA, of Synechococcus elongatus PCC 7942 is an active enzyme: a sequel to an old story.

Carbonic anhydrase (CA) EcaA of Synechococcus elongatus PCC 7942 was previously characterized as a putative extracellular α-class CA, however, its activity was never verified. Here we show that EcaA possesses specific CA activity, which is inhibited by ethoxyzolamide. An active EcaA was expressed in heterologous bacterial system, which supports the formation of disulfide bonds, as a full-length protein (EcaA+L) and as a mature protein that lacks a leader peptide (EcaA-L). EcaA-L exhibited higher specific activity compared to EcaA+L. The recombinant EcaA, expressed in a bacterial system that does not support optimal disulfide bond formation, exhibited extremely low activity. This activity, however, could be enhanced by the thiol-oxidizing agent, diamide; while a disulfide bond-reducing agent, dithiothreitol, further inactivated the enzyme. Intact E. coli cells that overexpress EcaA+L possess a small amount of processed protein, EcaA-L, whereas the bulk of the full-length protein resides in the cytosol. This may indicate poor recognition of the EcaA leader peptide by protein export systems. S. elongatus possessed a relatively low level of ecaA mRNA, which varied insignificantly in response to changes in CO2 supply. However, the presence of protein in the cells is not obvious. This points to the physiological insignificance of EcaA in S. elongatus, at least under the applied experimental conditions.

Draft Genome Sequences of Two Thermotolerant Cyanobacterial Strains Isolated from Hot Springs.

We report here two draft cyanobacterial genome sequences, those of Cyanobacterium aponinum IPPAS B-1201, isolated from a hot spring in the Turgen Gorge (Kazakhstan), and the uncharacterized cyanobacterium IPPAS B-1203, isolated from a hot spring in Karlovy Vary (Czech Republic). These two strains were deposited at the Collection of Microalgae (IPPAS) of the Timiryazev Institute of Plant Physiology.

Cyanofuels: biofuels from cyanobacteria. Reality and perspectives.

Cyanobacteria are represented by a diverse group of microorganisms that, by virtue of being a part of marine and freshwater phytoplankton, significantly contribute to the fixation of atmospheric carbon via photosynthesis. It is assumed that ancient cyanobacteria participated in the formation of earth's oil deposits. Biomass of modern cyanobacteria may be converted into bio-oil by pyrolysis. Modern cyanobacteria grow fast; they do not compete for agricultural lands and resources; they efficiently convert excessive amounts of CO2 into biomass, thus participating in both carbon fixation and organic chemical production. Many cyanobacterial species are easier to genetically manipulate than eukaryotic algae and other photosynthetic organisms. Thus, the cyanobacterial photosynthesis may be directed to produce carbohydrates, fatty acids, or alcohols as renewable sources of biofuels. Here we review the recent achievements in the developments and production of cyanofuels-biofuels produced from cyanobacterial biomass.

Cold-induced gene expression and ω(3) fatty acid unsaturation is controlled by red light in Synechocystis.

The expression of cold-induced genes, which are controlled by the cold sensor histidine kinase Hik33, and the formation of ω(3) polyunsaturated fatty acids are controlled by light in the cyanobacterium Synechocystis sp. PCC 6803. Cold-induced Hik33-dependent gene expression is initiated by red light (∼700nm), but not by blue or green light. Red light also turns on the ω(3) fatty acid desaturation. Different combinations of other wavelengths in red spectral region (635 and 726nm) had no effect on the red-light-activated cold-induced transcription or fatty acid desaturation. Therefore, the involvement of phytochrome-like photoreceptor(s), similar to phytochromes of higher plants, in this regulation was not confirmed. The absence of light-dependence of gene expression in the mutant cells deficient in Hik33 suggests the involvement of this histidine kinase in direct or mediated with red light regulation of cold responses in Synechocystis.

Light-dependent cold-induced fatty acid unsaturation, changes in membrane fluidity, and alterations in gene expression in Synechocystis.

Cold stress causes unsaturation of the membrane lipids. This leads to adjustment of the membrane fluidity, which is necessary for cold acclimation of cells. Here we demonstrate that the cold-induced accumulation of PUFAs in the cyanobacterium Synechocystis is light-dependent. The desA(-)/desD(-) mutant, that lacks the genes for Δ12 and Δ6 desaturases, is still able to adjust the fluidity of its membranes in spite of its inability to synthesize PUFAs and modulate the fatty acid composition of the membrane lipids under cold stress. The expression of cold-induced genes, which are controlled by the cold sensor histidine kinase Hik33, depends on the fluidity of cell membranes and it is regulated by light, though it does not require the activity of the photosynthetic apparatus. The expression of cold-induced genes, which are not controlled by Hik33, does not depend on the membrane fluidity or light. Thus, membrane fluidity determines the temperature dependence of the expression of cold-induced genes that are under control of the Hik33, which might be the sensor of changes in the membrane fluidity. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.