Claudio Sili

Exopolysaccharide-producing cyanobacteria and their possible exploitation: A review

Since the early 1950s, more than one hundred cyanobacterial strains,belonging to twenty different genera, have been investigated with regard tothe production and the released exocellular polysaccharides (RPS) into theculture medium. The chemical and rheological properties show that suchpolysaccharides are complex anionic heteropolymers, in about 80% casescontaining six to ten different monosaccharides and in about 90% casescontaining one or more uronic acids; almost all have non-saccharidiccomponents, such as peptidic moieties, acetyl, pyruvyl and/or sulphategroups. Based on such ingredients, cyanobacterial RPSs show promise asthickening or suspending agents, emulsifying or cation-chelating compoundsand the residual capsulated cyanobacterial biomass, following RPSextraction, could be an effective cation-chelating material. Indeed, wheneleven unicellular and filamentous RPS-producing cyanobacteria, selectedon the basis of the anion density of their RPSs and on the abundance oftheir outermost investments, were screened for their ability to removeCu2+ from aqueous solutions, a quick and most effective heavy metaladsorption was observed for the unicellular Cyanothece CE 4 and thefilamentous Cyanospira capsulata. These results suggest the possibilityto accomplish, through the exploitation of RPS-producing cyanobacteria,a multiproduct strategy to procure a wide range of biopolymers suited tovarious industrial applications, in addition to the residual biomass effectivein the recovery of heavy metals from polluted waters.

Assessment of the metal removal capability of two capsulated cyanobacteria, Cyanospira capsulata and Nostoc PCC7936

Two capsulated, exopolysaccharide-producing cyanobacteria, Cyanospira capsulata and Nostoc PCC7936, were tested with regard to their metal removal capability by using copper as model metal. The experiments, carried out with the sole cyanobacterial biomass suspended in distilled water and confined into small dialysis tubings, showed that C. capsulata biomass is characterized by the best efficiency in metal removal, with a qmax (maximum amount of copper removed per biomass unit) of 96 ± 2 mg Cu(II) removed per g of protein in comparison with the value of 79 ± 3 of Nostoc PCC7936 biomass. The experimental data obtained with both cyanobacterial biomass best fit the Langmuir sorption isotherm. The sorption of copper started from the first minutes of contact with the metal and attained the equilibrium state, when no more copper removal was evident, after 5 and 6 hours, for C. capsulata and Nostoc PCC7936, respectively. The best efficiency in Cu(II) removal was obtained at pH 6.1–6.2, while the presence of Mg2+ or Ca2+ reduced copper removal capability of both species to 60–70% of their qmax. The results showed that the biomass of C. capsulata and Nostoc PCC7936 possesses a high affinity and a high specific uptake for copper, comparable with the best performances shown by other microbial biomass, and suggest the possibility to use the capsulated trichomes of the two cyanobacteria for the bioremoval of heavy metals from polluted water bodies.

STUDIES ON EXOPOLYSACCHARIDE RELEASE BY DIAZOTROPHIC BATCH CULTURES OF CYANOSPIRA CAPSULATA

SummaryDiazotrophic batch cultures of Cyanospira capsulata producing large amounts of a soluble exopolysaccharide (EPS) were studied over a period of about 30 days under continuous illumination. The thickness of the capsule surrounding the trichomes remained almost the same throughout the growth phases and the EPS was continuously released into the medium at a rate which was roughly constant throughout the culture period. A mean EPS productivity of about 6 g m−2 day−1 was attained. Purified EPS samples exhibited a saccharidic composition consisting of four neutral sugars (glucose, mannose, fucose and arabinose) and galacturonic acid in a molar ratio of 1:1:1:1:2, respectively. The EPS was also characterized by the presence of pyruvic residues and by a protein content of about 2%. O-Acetyl groups and sulphate residues were not detected. The massive release of this polysaccharidic material into the liquid medium made the cultures progressively more viscous.

Glycogen and poly-β-hydroxybutyrate synthesis in Spirulina maxima

The effect of different growth conditions on the glycogen and poly-β-hydroxybutyrate (PHB) content of the cyanobacterium Spirulina maxima is described. Under photoautotrophic growth conditions without any nutrient limitation, S. maxima exhibited a glycogen content of between 7.1 and 10.7% of cell dry wt, whereas PHB was undetectable. When S. maxima was grown under mixotrophic conditions in the presence of acetate, the intracellular PHB concentration increased to more than 3% of dry wt, while glycogen content remained within the range of 5 to 6% of cell dry wt. Nitrogen starvation favoured glycogen accumulation (up to 60 to 70% of dry wt), while the PHB content remained low (up to 0.7% of dry wt), even after prolonged nitrogen starvation. Inhibition of protein synthesis, induced by addition of azaserine, led to the accumulation of glycogen (up to 52% of cell dry wt) but did not stimulate PHB synthesis. Under phosphorus-limited growth conditions, glycogen and PHB accumulated (up to 23% and 1.2% of cell dry wt, respectively) only after the exhaustion of intracellular phosphorus reserves. Shifting the culture from low to high light irradiance induced a rapid accumulation of glycogen (up to 34% of cell dry wt after 9 h) but did not induce PHB synthesis. Results are discussed in terms of the metabolic significance of PHB synthesis in cyanobacteria, and suggest that this polymer acts exclusively as a disposal mechanism to eliminate excess reducing equivalents.

Cyanospira rippkae and Cyanospira capsulata (gen. nov. and spp. nov.): new filamentous heterocystous cyanobacteria from Magadi lake (Kenya)

New filamentous heterocystous cyanobacteria were isolated from the alkaline soda lake Magadi in Kenya. The characteristics of the isolates are summarized and their taxonomic position discussed.Uniform attributes of the strains, grouped in two types, Mag II 702 and Mag I 504 are the following: helical structure of the trichomes, immotility, gas vacuolation, obligate autotrophy, nitrogen fixation under aerobic conditions, and closely similar fatty acid composition, including the uncommon cis-vaccenic acid.For these organisms the assignment to a new genus named Cyanospira is proposed with the species C. rippkae and C. capsulata, separated on the basis of structural, chemical and mean DNA-base composition. Type strains 702 and 504 will be deposited at the ATCC and PCC.

Biodiversity of Phototrophic Biofilms Dwelling on Monumental Fountains

Among the stone monumental assets, artistic fountains are particularly affected by microbial colonization due to constant contact with water, giving rise to biodegradation processes related with physical–chemical and aesthetical alterations. In this paper, we make an overview of reported biodiversity of the phototrophic patina developed in various fountains of Italy and Spain. The microbial composition of four fountains (two from Florence, Italy and two from Granada, Spain) was investigated using traditional and/or molecular techniques. The results indicated many common similarities with regard the phototrophic biodiversity for all the investigated fountains. Automated ribosomal RNA intergenic spacer analysis (ARISA), a molecular fingerprint tool, was used to examine the eubacterial and cyanobacterial community for two of the investigated fountains. The principal component analysis of ARISA profiles strengthens the results obtained by traditional methods and revealed separate clusters, as a consequence of the differences of micro-environmental conditions for each fountain.

Assessment of the potential of Nostoc strains from the Pasteur Culture Collection for the production of polysaccharides of applied interest

The exocellular polysaccharides released during thephotoautotrophic growth by 25 Nostoc strainsbelonging to the Pasteur Culture Collection (PCC) wereinvestigated with regard to their chemical andrheological properties in order to assess theirpotential for the production of polysaccharides ofindustrial interest.All the released polysaccharides (RPSs) were complexanionic heteropolymers, composed of a number ofmonosaccharides ranging from six to nine, alwaysincluding glucose and fucose, and most frequentlygalactose. Ribose, rarely reported to be present incyanobacterial RPSs, was found in seven polymers. TheRPSs were shown to possess an anionic charge due tothe presence of uronic acids and in most cases also ofpyruvil and sulphate groups. Moreover, a large numberof polymers showed the presence of significant amountsof constituents such as acetyl groups, peptidicmoieties and deoxysugars, that may contribute to thehydrophobicity of the macromolecules. The viscositydependence on the shear rate of aqueous solutions ofthe RPSs was, in three cases, comparable with that ofxanthan gum, and one RPS showed a very satisfactorystability of the viscosity over a wide range of pH,temperature and salinity values. The most promisingPCC Nostoc strains for the production of RPSsfor specific applications were thus singled out, basedon the characteristics of their polymers.

Response of an exopolysaccharide-producing heterocystous cyanobacterium to changes in metabolic carbon flux

The response of the exopolysaccharide-producing heterocystous cyanobacteriumCyanospira capsulata to changes in metabolic carbon flux was investigated to estimate the potential for improvement of the exopolysaccharide (EPS) yield. Carbon flux was altered by transferring the organism either to an argon atmosphere or to medium containing the nitrogen assimilation inhibitors L-methionine-D,L-sulfoximine (MSX), O-diazoacetyl-L-serine (AZAS) or D,L-7-azatryptophan (AZAT), or by adding glyoxylate, known to stimulate carbon metabolism. When carbon flux was modified by interfering with nitrogen metabolism, the concentration of total carbohydrates exceeded that of the control culture only in Ar- or AZAS-treated cell suspensions, but this difference was mainly due to enhancement of the quantity of bound carbohydrates. On the other hand, when carbon flux was modified by a single addition of glyoxylate (30 mM) or by daily additions of 10 mM glyoxylate without interfering with nitrogen metabolism, carbohydrate release into the medium was stimulated markedly; after 5 days of growth in the presence of the organic compound, the concentration of EPS was 43 % higher than in the control culture. The results demonstrate that, with enhanced carbon flux, the excess carbon is preferentially channeled byC. capsulata cells into the synthesis of an overflow product like EPS, whereas, with mere diversion of carbon flux from the process of nitrogen assimilation, the synthesis of carbon reserves is more heavily favoured.

Hydrogen production by immobilized cells. III: Prolonged and stable H2 photoevolution by rhodopseudomonas Palustris in light-dark cycles

Considerable progress in the photobiological hydrogen production process by agar-entrapped resting cells of Rhodopseudomonas palustris in light-dark cycles was achieved by improving the substrate diffusion, by preventing a high uptake hydrogenase activity and by eliminating nitrogen starvation, which induces deleterious effects on nitrogenase function. Keeping to these directions a stable rate of H2 photoevolution was maintained for 100 days, when the experiment was terminated. The thermodynamic conversion efficiency of the process reached a value of about 3.4%.

Germination of desiccated aged akinetes of alkaliphilic cyanobacteria

Morphological and biochemical changes associated with synchronous germination of mature, aged and desiccated akinetes of two alkaliphilic cyanobacteria, Cyanospira rippkae and Cyanospira capsulata, are described. Akinetes of both strains proved to be highly resistant to desiccation, being able to germinate, in the presence of either N2 or nitrate as nitrogen source, with a germination frequency of more than 90% after seven years of storage in a dried state. The first cell division occurred after 8–10 h of incubation, thereafter the germlings of the two strains followed a different pattern of cell differentiation. Heterocysts were first noted, in a terminal position, at 16–18 h in three-celled germlings of C. capsulata and at 21–24 h in C. rippkae, when germlings were at least seven cells in length. Akinetes of both species possessed, on a per cell basis, almost identical amounts of all photosynthetic pigments but, under nitrogen fixing conditions, photosynthetic activity (oxygen evolution) was detected only after new proteins had been synthesized, before a functional heterocyst was developed and while total nitrogen remained constant. With energy provided by aerobic respiration, a wide range of intracellular amino acids characteristic of proteins was utilised to sustain the new protein synthesis. The end of this biosynthetic activity coincided with the timing of the first cell division. From this stage on, no changes in protein concentration occurred until mature heterocysts were developed. In the presence of nitrate, no significant changes in the major germination events were observed.