Cyanobacterial extracellular polymeric substances (EPS) are mainly composed of high-molecular-mass heteropolysaccharides, with variable composition and roles according to the microorganism and the environmental conditions. The number of constituents - both saccharidic and nonsaccharidic - and the complexity of structures give rise to speculations on how intricate their biosynthetic pathways could be, and how many genes may be involved in their production. However, little is known regarding the cyanobacterial EPS biosynthetic pathways and regulating factors. This review organizes available information on cyanobacterial EPS, including their composition, function and factors affecting their synthesis, and from the in silico analysis of available cyanobacterial genome sequences, proposes a putative mechanism for their biosynthesis.

Complexity of cyanobacterial exopolysaccharides: composition, structures, inducing factors and putative genes involved in their biosynthesis and assembly

Dating from the Pre-Cambrian era, cyanobacteria have a long history of adaptation to the Earth's environment. By evolving oxygen via photosynthetic reactions similar to those of plants and green algae, these prokaryotes were essential to the evolution of the present biosphere. They continue to make a large contribution to the equilibrium of the Earth's atmosphere by production oxygen and removing carbon dioxide. To survive in extreme or variable environments, cyanobacteria have developed specific regulatory systems, in addition to more general mechanisms equivalent to those of other prokaryotes or photosynthesis eukaryotes. Specific regulatory systems control the differentiation of specialized nitrogen-fixing cells and of cell types facilitating the dispersion of species. In the past decade, considerable progress has been made towards understanding the expression of the cyanobacterial genome in response to variations in the intensity and spectral quality of incident light and in response to nutritional conditions, especially carbon, nitrogen and sulphur sources. These studies have provided insights into the relationships between carbon and nitrogen intermediary metabolism, and a start towards understanding of the interconnected pathways which lead from the perception of environmental signals to the regulation of enzyme activities and gene expression. Cyanobacterial regulatory mechanisms share common features with those of other prokaryotes, but are unique since these essentially photo-autotrophic organisms must maintain a proper cellular C/N balance, in spite of dailty variations in incident light. Thus an appropriate coordination between photosynthesis and other metabolic processes must be achieved through control of the catalytic activity of key enzymes by reducing equivalents and ATP produced by photosynthetic or respiratory electron transport. Recently discovered kinases/phosphatases act by post-translational modification of specific proteins which probably act as signal transducers or modulators of gene expression in a manner similar to the well-known two-component regulatory systems described in other bacteria. In this overview, we present our current knowledge on the molecular aspects of the biology of cyanobacteria, as well as on their mechanisms of resistance to metal ions and their responses to metabolic stress.

Adaptation of cyanobacteria to environmental stimuli: new steps towards molecular mechanisms

Arsenic ions, frequently present as environmental pollutants, are very toxic for most microorganisms. Some microbial strains possess genetic determinants that confer resistance. In bacteria, these determinants are often found on plasmids, which has facilitated their study at the molecular level. Bacterial plasmids conferring arsenic resistance encode specific efflux pumps able to extrude arsenic from the cell cytoplasm thus lowering the intracellular concentration of the toxic ions. In Gram-negative bacteria, the efflux pump consists of a two-component ATPase complex. ArsA is the ATPase subunit and is associated with an integral membrane subunit, ArsB. Arsenate is enzymatically reduced to arsenite (the substrate of ArsB and the activator of ArsA) by the small cytoplasmic ArsC polypeptide. In Gram-positive bacteria, comparable arsB and arsC genes (and proteins) are found, but arsA is missing. In addition to the wide spread plasmid arsenic resistance determinant, a few bacteria confer resistance to arsenite with a separate determinant for enzymatic oxidation of more-toxic arsenite to less-toxic arsenate. In contrast to the detailed information on the mechanisms of arsenic resistance in bacteria, little work has been reported on this subject in algae and fungi.

Resistance to arsenic compounds in microorganisms

Cell coloration changes from normal blue-green to yellow or yellow-green when the cyanobacterium Synechococcus sp. strain PCC 7942 is deprived of an essential nutrient. We found that this bleaching process (chlorosis) in cells deprived of sulfur (S) was similar to that in cells deprived of nitrogen (N), but that cells deprived of phosphorus (P) bleached differently. Cells divided once after N deprivation, twice after S deprivation, and four times after P deprivation. Chlorophyll (Chl) accumulation stopped almost immediately upon N or S deprivation but continued for several hours after P deprivation. There was no net Chl degradation during N, S, or P deprivation, although cellular Chl content decreased because cell division continued after Chl accumulation ceased. Levels of the light-harvesting phycobiliproteins declined dramatically in a rapid response to N or S deprivation, reflecting an ordered breakdown of the phycobilisomes (PBS). In contrast, P-deprived cultures continued to accumulate PBS for several hours. Whole PBS were not extensively degraded in P-deprived cells, although the PBS contents of P-deprived cells declined because of continued cell division after PBS accumulation ceased. Levels of mRNAs encoding PBS polypeptides declined by 90 to 95% in N- or S-deprived cells and by 80 to 85% in P-deprived cells. These changes in both the synthesis and stability of PBS resulted in a 90% decline in the PC/Chl ratio of N- or S-deprived cells and a 40% decline in the PC/Chl ratio of P-deprived cells. Therefore, although bleaching appears to be a general response to nutrient deprivation, it is not the same under all nutrient-limited conditions and is probably composed of independently controlled subprocesses.

Chlorosis induced by nutrient deprivation in Synechococcus sp. strain PCC 7942: not all bleaching is the same.

Kinetics of nutrient-limited transport and microbial growth.

The phosphorus contents of acid-soluble pools, lipid, ribonucleic acid, and acid-insoluble polyphosphate were lowered in Synechococcus in proportion to the reduction in growth rate in phosphate-limited but not in nitrate-limited continuous culture. Phosphorus in these cell fractions was lost proportionately during progressive phosphate starvation of batch cultures. Acid-insoluble polyphosphate was always present in all cultural conditions to about 10% of total cell phosphorus and did not turn over during balanced exponential growth. Extensive polyphosphate formation occurred transiently when phosphate was given to cells which had been phosphate limited. This material was broken down after 8 h even in the presence of excess external orthophosphate, and its phosphorus was transferred into other cell fractions, notably ribonucleic acid. Phosphate uptake kinetics indicated an invariant apparent Km of about 0.5 μM, but Vmax was 40 to 50 times greater in cells from phosphate-limited cultures than in cells from nitrate-limited or balanced batch cultures. Over 90% of the phosphate taken up within the first 30 s at 15°C was recovered as orthophosphate. The uptake process is highly specific, since neither phosphate entry nor growth was affected by a 100-fold excess of arsenate. The activity of polyphosphate synthetase in cell extracts increased at least 20-fold during phosphate starvation or in phosphate-restricted growth, but polyphosphatase activity was little changed by different growth conditions. The findings suggest that derepression of the phosphate transport and polyphosphate-synthesizing systems as well as alkaline phosphatase occurs in phosphate shortage, but that the breakdown of polyphosphate in this organism is regulated by modulation of existing enzyme activity.

John F. Grillo

Papers

Regulation of Phosphate Accumulation in the Unicellular Cyanobacterium Synechococcus