Inorganic carbon acquisition systems in cyanobacteria

This minireview focuses on the mechanism of inorganic carbon uptake in cyanobacteria and in particular the two CO₂-uptake systems and two bicarbonate transporters recently identified in Synechocycstis PCC 6803, and their presence in other cyanobacterial strains. This revised version was published online in August 2006 with corrections to the Cover Date.     

Physiological characteristics of cyanobacteria in pulp and paper waste-treatment systems

An investigation was made into the physiological characteristics of the extensive cyanobacterial communities in pulp and paper secondary waste-treatment systems, including the capacity of isolates to biodegrade organic contaminants in these systems. Although pulp and paper waste-treatment systems were found to be severely light-limited, photosynthesis-irradiance curves indicated that shade-adapted cyanobacterial communities could fix conspicuous amounts of inorganic carbon via photosynthesis. Of 21 cyanobacterial strains isolated from pulp and paper waste-treatment systems located in 4 countries, all except one were capable of glucose uptake in the light, and 19 also showed uptake in the dark. In the 6 species tested, glucose and acetate addition stimulated growth in low light in all except one species. Aphanocapsa rivularis grew equally well on glucose and acetate in the dark but Pseudanabaena sp. grew well on acetate, and minimally on glucose. Growth stimulation by glucose and acetate in these two strains was greater in low than in high light. Pseudanabaena sp. and Phormidium animale both accumulated 2,4-dichlorophenol and 3-chlorobenzoate but showed minimal mineralization to CO₂. None of the four species tested could accumulate or degrade phenol or dichloroacetate. It is concluded that, depending on the light conditions, cyanobacteria contribute organic carbon in photosynthesis, and/or remove small organic molecules during mixotrophic and heterotrophic growth but are not important degraders of contaminants in these waste treatment systems.     

The occurrence of cyanobacteria in pulp and paper waste-treatment systems

Pulp and paper secondary waste-treatment systems in Brazil, Canada, New Zealand, and the U.S.A. contained dynamic cyanobacterial communities, some of which exceeded heterotrophic bacterial biomass. No other viable photoautotrophic populations were detected in the ponds. Regardless of geographical location, Oscillatoriales including Phormidium, Geitlerinema, and Pseudanabaena were the dominant taxa. As well, Chroococcus (Chroococcales) was an important genus in Brazil and New Zealand. The possible impact of cyanobacteria on waste-treatment efficiency deserves further study given their large biomass and diverse metabolic characteristics.     

Special roles for efflux systems in iron homeostasis of non-siderophore-producing cyanobacteria

In oligotrophic oceans, low bioavailability of Fe is a key factor limiting primary productivity. However, excessive Fe in cells leads to the Fenton reaction, which is toxic to cells. Cyanobacteria must strictly maintain intracellular Fe homeostasis. Here, we knocked out a series of genes encoding efflux systems in Synechocystis sp. PCC 6803, and found eight genes that are required for high Fe detoxification. Unexpectedly, the HlyBD-TolC efflux system plays an important role in the adaptation of Synechocystis under Fe-deficient conditions. Mutants of HlyD and TolC grew worse than the wild-type strain under low-Fe conditions and showed significantly lower intracellular Fe contents than the wild-type strain. We excluded the possibility that the low Fe sensitivity of the HlyBD-TolC mutants was caused by a loss of the S-layer, the main extracellular protein secreted via this efflux system. Inactivation of the HlyD protein influenced type IV pili formation and direct inactivation of type IV pili related genes affected the adaptation to low-Fe conditions. HlyBD-TolC system is likely involved in the formation of type IV pili and indirectly influenced Fe acquisition. Our findings suggest that efflux system in non-siderophore-producing cyanobacteria can facilitate Fe uptake and help cells adapt to Fe-deficient conditions via novel pathways.     

Sensing the light: photoreceptive systems and signal transduction in cyanobacteria

Photosynthetic prokaryotes have highly developed abilities to detect and react to environmental signals. Light sensing is one of the most important capabilities of organisms that use light for photosynthesis and photomorphogenesis. This review addresses photoreception in cyanobacteria from the perception of light through the physiological responses observed in response to light-dependent signalling. Recent progress made in our understanding of the structure and function of photosensory receptors and their downstream effector molecules is discussed.     

Geosmin and 2-methylisoborneol from cyanobacteria in three water supply systems

The changes in populations of Staphylococcus aureus, Bacillus subtilis, Salmonella typhimurium, Klebsiella pneumoniae, Agrobacterium tumefaciens, Rhizobium meliloti, and Saccharomyces cerevisiae were measured after their introduction into samples of sewage, lake water, and soil. Enumeration of small populations was possible because the strains used were resistant to antibiotics in concentrations and combinations such that few species native to these ecosystems were able to grow on agar containing the inhibitors. Fewer than 2 cells per ml of sewage or lake water and 25 cells per g of soil could be detected. A. tumefaciens and R. meliloti persisted in significant numbers with little decline, but S. aureus, K. pneumoniae, S. typhimurium, S. cerevisiae, and vegetative cells of B. subtilis failed to survive in samples of sewage and lake water. In sterile sewage, however, K. pneumoniae, B. subtilis, S. typhimurium, A. tumefaciens, and R. meliloti grew; S. cerevisiae populations were maintained at the levels used for inoculation; and S. aureus died rapidly. In sterile lake water, the population of S. aureus and K. pneumoniae and the number of vegetative cells of B. subtilis declined rapidly, R. meliloti grew, and the other species maintained significant numbers with little or a slow decline. The populations of S. aureus, K. pneumoniae, A. tumefaciens, B. subtilis, and S. typhimurium declined in soil, but the first four species grew in sterile soil. It is suggested that some species persist in environments in which they are not indigenous because they tolerate abiotic stresses, do not lose viability readily when starved, and coexist with antagonists. The species that fails to survive need only be affected by one of these factors.     

Transport of D-glucose and 3-O-methyl-D-glucose in the cyanobacteria Aphanocapsa 6714 and Nostoc strain Mac.

1. The cyanobacterium Aphanocapsa 6714 which grow in the dark on D-glucose, will take up D-glucose and the analogue 3-O-methyl-D-glucose; uptake of each of these compounds was inhibited competitively by the other and by 6-deoxy-D-glucose. 2. This cyanobacterium accumulated 3-O-methyl-D-glucose up to 100-fold relative to the medium but did not modify or metabolize it to a significant degree. 3. Intracellular 3-O-methyl-D-glucose was rapidly displaced from Aphanocapsa 6714 by exogenous D-glucose and 3-O-methyl-D-glucose. 4. Although not characterized to the same extent, D-glucose and 3-O-methyl-D-glucose uptake by Nostoc strain Mac, another cyanobacterium capable of growth in the dark on D-glucose, was similar. 5. Other cyanobacteria that do not grow on D-glucose take up this compound at much lower rates which were unaffected by analogues of D-glucose that greatly reduced carbohydrate uptake by Aphanocapsa 6714 and Nostoc strain Mac. 6. It is therefore proposed that Aphanocapsa 6714 and Nostoc strain Mac possess a mechanism for the active transport of D-glucose. The absence of this transport mechanism is suggested as the reason why other strains fail to grow in the dark on this substrate. These latter organisms are therefore naturally cryptic with respect to D-glucose as a growth substrate.     

Transport and accumulation of bloom-forming cyanobacteria in a large, mid-latitude lake: the gyre-Microcystis hypothesis

 Toxic cyanobacterial blooms have occurred in the near-shore waters of the North Basin of Lake Biwa, Japan, since 1994, and have been attributed to deterioration of water quality in the enriched littoral zone of the lake. From 1997 onwards, the bloom-forming cyanobacteria have been observed with increasing frequency in the deep offshore waters of the North Basin. In the present study, we examined the mechanisms responsible for these bloom populations in the main body of the lake. Specifically, we addressed the hypothesis that buoyant, nutrient-replete colonies of cyanobacteria are generated inshore, are advected offshore by large-scale horizontal transport processes, and subsequently accumulate in the downwelling center of large surface gyres that characterize the overall circulation pattern in the epilimnion of the North Basin. Diel variations of Microcystis biomass at the center and the edge of the Lake Biwa gyre were monitored at 6-h intervals on August 23–24, 2000, and the horizontal distribution of buoyant Microcystis was determined on October 6. The hydrodynamic structure of the first gyre was determined over the preceding 2 days by an on-board Acoustic Doppler Current Profiler (ADCP). The gyre was characterized by a counterclockwise horizontal current that could potentially advect material large distances offshore, a downwelling current near the center of the gyre, and an upwelling current at the edge of the gyre, caused by the radial pressure gradients. The biomass of Microcystis near the water surface was greater at the center than at the edge of the gyre, and the biomass at 5 m depth at the edge of the gyre was greater than that at the water surface or at the thermocline near the edge of the gyre. The results are consistent with the gyre-Microcystis hypothesis, and show the potential for accumulation of large concentrations of cyanobacteria in deep offshore lake environments that are normally considered unsuitable for cyanobacterial blooms. Received: July 16, 2001 / Accepted: March 6, 2002     

Transport systems encoded by bacterial plasmids

A variety of bacterial functions are encoded on plasmids, extrachromosomal elements. Examples of plasmid-borne functions are antibiotic production and resistance, degradation of recalcitrant chemicals, virulence factors, and plant symbiotic properties. Several transport systems with diverse functions have recently been found to be carried on plasmids. These systems serve to either accumulate or extrude a compound from a cell. The focus of this review is to present a survey on several of these novel plasmid-borne transport systems emphasizing functions, components, and molecular genetics.     

Ploidy in cyanobacteria

A recently developed real-time PCR method for the determination of genome copy numbers was optimized for the application to cyanobacteria. Three species were chosen to represent a fresh water species, a salt water species, and two strains of a widely used laboratory species. Synechococcus PCC 7942 and Synechococcus WH7803 were found to contain 3-4 genome copies per cell and are thus oligoploid, confirming earlier publications. In contrast, Synechocystis PCC 6803 is highly polyploid. The motile wild-type strain contains 218 genome copies in exponential phase and 58 genome copies in linear and in stationary growth phase. The GT wild-type strain contains 142 genome copies in exponential phase and 42 genome copies in linear and stationary growth phase. These are the highest numbers found for any cyanobacterial species. Notably these values are much higher than the value of 12 genome copies published for the 'Kazusa' strain more than 20 years ago. The results reveal that for Synechocystis PCC 6803 strain differences exist and that the ploidy level is highly growth phase-regulated. A compilation of the ploidy levels of all investigated cyanobacterial species gives an overview of the genome copy number distribution and shows that monoploid, oligoploid, and polyploid cyanobacteria exist.     

The role of systems biology in developing non-model cyanobacteria as hosts for chemical production

1. Download : Download high-res image (127KB)
2. Download : Download full-size image

     

Lactate dehydrogenases in cyanobacteria

NAD-linked lactate dehydrogenases specific for the D- and L-lactate have been demonstrated in a number of strains of unicellular cyanobacteria. The D-lactate dehydrogenase of one strain (Synechococcus 6716) was partially purified and its properties were studied. The enzyme has a molecular weight of ca. 115000-120000, is highly specific, autooxidizable, and susceptible to inhibition by iodoacetamide, oxamate and ATP. The possible physiological functions of the enzyme in the metabolism of the organism were investigated. D-lactate carbon was incorporated in cell material during photosynthetic growth with CO2, but lactate was not used as sole source for carbon for photosynthetic or chemosynthetic development. D-lactate and pyruvate were oxidized aerobically in the dark by resting cell suspensions with the assimilation mainly of the C2 and the C3 carbon atoms. In the oxidation of lactate, acetate was excreted into the medium. No fermentation of glucose was found, but a small amount of D-lactate was detected as a product of endogenous dark metabolism of the cell. All enzymes required for the production of lactate from glucose and from glycogen were found in exponentially growing cells, but the activity of some key enzymes was low or undetectable in old cultures.     

Iron deprivation in cyanobacteria

Iron is an essential component of electron transport in almost all living organisms. It is particularly important to phototrophs like cyanobacteria because 22–23 irons are required for a complete functional photosynthetic apparatus. Since the low solubility of Fe⁺⁺⁺ above neutral pH in oxic ecosystems severely limits the biological availability of iron to aquatic microorganisms, cyanobacteria and other microbes have developed a number of responses to cope with iron deficiency. Cyanobacterial responses to iron stress include the synthesis of an efficient, siderophore-based system to scavenge iron and the substitution of ferredoxin with flavodoxin. An additional response in cyanobacteria involves the alteration of the light-harvesting apparatus that includes the appearance of a new, iron-stress-induced, photosystem II, chlorophyll-binding protein. Although cytochromec-553 has a potential non-iron-containing replacement in plastocyanin, a copper-containing protein, iron stress appears to favor the utilization of cytochromec-553 because siderophores also bind copper and form a complex that is excluded from the cell.This paper is intended primarily as a review of molecular and physiological responses of actively growing cyanobacterial cultures to conditions of iron stress, where iron is present but essentially insoluble, and to differentiate these responses from iron starvation, where the amount of iron in the system is not sufficient for cell growth.     

Hydrogenase activities in cyanobacteria

In the unicellular Anacystis nidulans, the expression of both the H2-uptake (with phenazine methosulfate or methylene blue as the electron acceptor) and H2-evolution (with methyl viologen reduced by Na2S2O4) was dependent on Ni in the culture medium. In extracts from Anacystis and Anabaena 7119, H2-evolution and uptake activities were strongly inhibited by Cu2+, p-chloromercuribenzoate and HgCl2 suggesting that at least one functional SH-group is involved in catalysis by hydrogenase. Extracts from the N2-fixing Anabaena 7119 contained two different hydrogenase fractions which could be separated by chromatography on DE-52 cellulose using a linear NaCl concentration gradient. The fraction eluting with 0.13 M NaCl from the column catalyzed only the uptake of H2 with methylene blue as the electron acceptor but virtually not the evolution of H2 ("uptake" hydrogenase fraction). The fraction eluting at a NaCl strength of 0.195 M catalyzed both H2-uptake with methylene blue and H2-evolution with reduced methyl viologen ("reversible" hydrogenase fraction). Growth under anaerobic conditions drastically enhanced the activity levels of the "reversible" but not of the "uptake" hydrogenase fraction. The "uptake" hydrogenase but not the "reversible" protein was activated by reduced thioredoxin. It is suggested that thioredoxin activates the H2-uptake by the membrane-bound "uptake" hydrogenase also in intact cells. The occurrence of the number of hydrogenases in cyanobacteria will be reevaluated.     

Glycolate metabolism in cyanobacteria

A comparative analysis of glycolate excretion in 11 cyanobacteria showed that 8 strains, although grown and assayed in air, excreted glycolate. The largest quantities were excreted by the filamentous strains Plectonema boryanum 73110 and Anabaena cylindrica (Lemm). The carbon lost by excretion was at most 9% of the net fixed carbon in air for heterocystous cyanobacteria but increased (up to 60%) in some strains under a high pO₂ (0.03 kPa CO₂ in pure O₂). A. cylindrica excreted glycolate at a maximum level of 2 and 10 μmol (mg chl a )⁻¹ h⁻¹ in air and at high pO₂, respectively. The excretion continued for several hours. Increases in light intensity and pO₂ and a shift in pH from 7 to 9 increased the amount of glycolate excreted. A. cylindrica also showed the most O₂-sensitive fixation of CO₂. In vitro activity of phosphoglycolate phosphatase (EC 3.1.3.18) was found in all strains tested, with the highest activities noted for Gloeobacter violaceus 7.82 and Gloeothece 6909 and for young cultures of A. cylindrica . The lowest activities were found in Anabaena 7120 and Anacystis nidulans 625, strains excreting no or only minor quantities of glycolate.