Induction of mycosporine-like amino acids (MAAs) in cyanobacteria by solar ultraviolet-B radiation

Three filamentous and heterocystous N₂-fixing cyanobacteria, Anabaena sp., Nostoc commune and Scytonema sp. were tested for the presence of ultraviolet-absorbing mycosporine-like amino acids (MAAs) and their induction by solar ultraviolet-B (UV-B) radiation. High performance liquid chromatographic (HPLC) studies revealed the presence of only one type of MAAs in all three cyanobacteria, that was identified as shinorine, a bisubstituted MAA containing both glycine and serine groups having an absorption maximum at 334 nm and a retention time of around 2.8 min. There was a circadian induction in the synthesis of MAAs when the cultures were exposed to mid-latitude solar radiation (Playa Unión, Rawson, Chubut, Patagonia, Argentina) for 3 days, 4–6th February, 2000. Solar radiation was measured by an ELDONET (European Light Dosimeter Network) filter radiometer permanently installed on the roof of the Estación de Fotobiología Playa Unión (43°18′ S; 65°03′ W). The maximum irradiances were around 450–500, 45–50 and 1.0–1.2 W m⁻² for PAR (photosynthetic active radiation), UV-A (ultraviolet-A) and UV-B (ultraviolet-B), respectively. PAR and UV-A had no significant impact on MAA induction while UV-B induced the synthesis of shinorine in all three cyanobacteria. Shinorine was found to be induced mostly during the light period. During the dark period the concentration stayed almost constant. In addition to shinorine, another unidentified, water-soluble, brownish compound with an absorption maximum at 315 nm was found to be induced by UV-B only in Scytonema sp. and released into the medium. This substance was neither found in Anabaena sp. nor in Nostoc commune. Judging from the results, the studied cyanobacteria may protect themselves from deleterious short wavelength radiation by their ability to synthesize photoprotective compounds in response to UV-B radiation.     

UV-B-induced synthesis of mycosporine-like amino acids in three strains of Nodularia (cyanobacteria)

Three filamentous and heterocystous cyanobacterial strains of Nodularia, Nodularia baltica, Nodularia harveyana and Nodularia spumigena, have been tested for the presence and induction of ultraviolet-absorbing/screening mycosporine-like amino acids (MAAs) by simulated solar radiation in combination with 395 (receiving photosynthetically active radiation (PAR) only), 320 (receiving PAR + UV-A) and 295 (receiving PAR + UV-A + UV-B) nm cut-off filters. Absorption spectroscopic analyses of the methanolic extracts of samples revealed a typical MAA peak at 334 nm in all three cyanobacteria. Specific contents of MAAs had a pronounced induction in the samples covered with 295 nm cut-off filters after 72 h of irradiation. In comparison, there was little induction of MAAs in the samples covered by 395 and 320 nm cut-off filters. High performance liquid chromatographic (HPLC) studies revealed the presence of two types of MAAs in all three cyanobacteria, which were identified as shinorine and porphyra-334, both absorbing maximally at 334 nm. The occurrence of porphyra-334 is rare in cyanobacteria. Specific content of both shinorine and porphyra-334 were induced remarkably only in the samples covered with 295 nm cut-off filters. The results indicate that in comparison to UV-A and PAR, UV-B is more effective in eliciting MAAs induction in the studied cyanobacteria.     

Protective effects of mycosporine-like amino acids of Synechocystis sp. PCC 6803 and their partial characterization

In this work, mycosporine-like amino acids (MAAs) of Synechocystis sp. PCC 6803 were characterized and were investigated on UV induction and protective ability. High performance liquid chromatographic (HPLC) studies revealed three major compounds in the MAAs. By UV absorption and mass spectra analysis, one of the compounds was tentatively identified as mycosporine-tau (M-tau). One novel compound similar to usujirene was tentatively named as dehydroxylusujirene, and the other novel compound was named as M-343 according to its absorption maximum. In vivo experiments indicated that M-tau was induced by both UV-A and UV-B, while dehydroxylusujirene and M-343 were only induced by UV-A, suggesting that different chromophores were involved in MAAs synthesis in Synechocystis sp. PCC 6803. It was also indicated that M-343 could be photochemically synthesized from some precursors. Under both UV and oxidation stresses, M-343 was more stable than dehydroxylusujirene and M-tau. Considering the reaction with H2O2, M-tau and dehydroxylusujirene might be potential antioxidants in reaction with physiological reactive oxygen species in vivo. In protection experiments, the MAAs exhibited efficient protective ability towards UV-B and H2O2 stresses, with maximal protection rates of 30% and 21.5%, respectively. These results indicate that the MAAs in Synechocystis sp. PCC 6803 act as both UV-screen and antioxidant.     

Mycosporine-like Amino Acids in Freshwater Copepods: Potential Sources and Some Factors That Affect Their Bioaccumulation

Mycosporine-like amino acids (MAAs) are ubiquitous photoprotective compounds in aquatic environments. MAAs are synthesized by a wide variety of organisms (i.e. bacteria, fungi and algae) and their production is photoinducible by ultraviolet radiation (UVR) (280–400 nm) and/or photosynthetically active radiation (400–750 nm). Most animals however, are unable to synthesize MAAs and must acquire these compounds through their diet or from symbiotic organisms. In this paper, we investigate the possible sources of MAAs and factors (temperature and initial MAA concentration) that may affect their bioaccumulation in freshwater copepods. We found that MAA accumulation may occur even if the copepods are cultured on a MAA-free diet. In addition, we found that the bacteriostatic antibiotic, chloramphenicol, inhibits the bioaccumulation of MAAs. These two pieces of evidence suggest that the source of MAAs in these copepods may be prokaryotic organisms in close association with the animals. The two factors investigated in this study, temperature and initial MAA concentrations, were found to affect the rates at which MAAs are accumulated. Temperature had positive effects on both uptake and elimination rates. On the other hand, the rate of uptake decreased at the highest assayed initial MAA concentration, probably because the concentration of MAAs was already close to saturation.     

Electrospray ionization tandem mass spectrometric and electron impact mass spectrometric characterization of mycosporine-like amino acids

Positive-ion mass spectral fragmentations of seven mycosporine-like amino acids (MAAs) are reported and discussed. The MAAs studied are small compounds composed of a cycloheximine ring substituted with amino acid or amino alcohol units. Techniques used include electron impact (EI) and electrospray ionization (ESI) with tandem mass spectrometry (MS/MS). ESI-MS/MS showed unusual small radical losses, generally resulting from the loss of a methyl group with the exception of shinorine and porphyra for which the initial losses were 30 and 44 Da, respectively. As expected from structural similarities, porphyra, shinorine and palythinol displayed similar fragmentation patterns, while palythenic acid and palythene fragmented in a similar manner. Overall, the ESI-MS/MS fragmentations at m/z <200 exhibited a distinctive pattern for all seven MAAs with characteristic ions at m/z 137, 168, 186, and 197 or 199. Several ions were observed for each of the MAAs analyzed, and together provide a useful and potentially diagnostic pattern for identification of MAAs and as an aid in structure elucidation of novel MAAs. For GC/EI-MS analysis, trimethylsilyl (TMS) derivatives were made. The EI-MS fragmentation patterns of TMS-MAAs showed many features typical of TMS-derivatized alpha-amines. The precursor TMS-MAA ion was not detected, but a [M-90](+ radical) ion was the highest-mass intense peak observed for palythine, palythinol and shinorine, while palythene gave a [M-116](+ radical) ion. Besides determining the number of acidic hydrogens, EI-MS of TMS-derivatized MAAs will aid in structure elucidation of novel MAAs.     

Effect of light quality on the accumulation of photosynthetic pigments, proteins and mycosporine-like amino acids in the red alga Porphyra leucosticta (Bangiales, Rhodophyta)

The effect of different light qualities (white, blue, green, yellow and red light) on photosynthesis, measured as chlorophyll fluorescence, and the accumulation of photosynthetic pigments, proteins and the UV-absorbing mycosporine-like amino acids (MAAs) was studied in the red alga Porphyra leucosticta. Blue light promoted the highest accumulation of nitrogen metabolism derived compounds i.e., MAAs, phycoerythrin and proteins in previously N-starved algae after seven days culture in ammonium enriched medium. Similar results were observed in the culture under white light. In contrast, the lowest photosynthetic capacity i.e., lowest electron transport rate and lowest photosynthetic efficiency as well as the growth rate were found under blue light, while higher values were found in red and white lights. Blue light favored the accumulation of the MAAs porphyra-334, palythine and asterina-330 in P. leucosticta. However, white, green, yellow and red lights favored the accumulation of shinorine. The increase of porphyra-334, palythine and asterina-330 occurred in blue light simultaneous to a decrease in shinorine. The accumulation of MAAs and other nitrogenous compounds in P. leucosticta under blue light could not be attributed to photosynthesis and the action of a non-photosynthetic blue light photoreceptor is suggested. A non-photosynthetic photoreceptor could be also involved in the MAAs interconversion pathways in P. leucosticta.     

Characterization of Truncated <Emphasis Type="Italic">dsz</Emphasis> Operon Responsible for Dibenzothiophene Biodesulfurization in <Emphasis Type="Italic">Rhodococcus</Emphasis> sp. FUM94

Numerous desulfurizing bacteria from the Rhodococcus genus harbor conserved dsz genes responsible for the degradation of sulfur compounds through 4S pathway. This study describes a newly identified desulfurizing bacterium, Rhodococcus sp. FUM94, which unlike previously identified strains encodes a truncated dsz operon. DNA sequencing revealed a frameshift mutation in the dszA gene, which led to an alteration of 66 amino acids and deletion of other C-terminal 66 amino acids. The resulting DszA polypeptide was shorter than DszA in Rhodococcus sp. IGTS8 reference strain. Despite the truncation, desulfurizing activity of the operon was observed and attributed to the removal of an overlap of dszA and dszB genes, and lack of active site in the altered region. Desulfurization experiments resulted in specific production rate of 6.3 mmol 2-hydroxy biphenyl (kgDCW)^?1 h^?1 at 2 g l^?1 biocatalyst concentration and 68.8% biodesulfurization yield at 20 g l^?1 biocatalyst concentration, both at 271 μM dibenzothiophene concentration which is comparable to similar wild-type biocatalysts.     

Polychromatic action spectrum for the induction of a mycosporine-like amino acid in a rice-field cyanobacterium, Anabaena sp

A polychromatic action spectrum for the induction of an ultraviolet-absorbing/screening mycosporine-like amino acid (MAA) has been determined in a filamentous and heterocystous nitrogen-fixing rice-field cyanobacterium, Anabaena sp. High-performance liquid chromatographic (HPLC) studies revealed the presence of only one type of MAA, which was identified as shinorine, a bisubstituted MAA containing both glycine and serine groups having a retention time at 2.8 min and an absorption maximum at 334 nm. Exposure of cultures to simulated solar radiation in combination with various cut-off filters (WG 280, 295, 305, 320, 335, 345, GG 400, 420, 455, 475, OG 515, 530, 570, RG 645, 665 and a broad-band filter, UG 11) clearly revealed that the induction of the MAA takes place only in the UV range. Photosynthetic active radiation (PAR) had no significant impact on MAA induction. The ratio of the absorption at 334 nm (shinorine) to 665 nm (chlorophyll a) and the action spectrum also showed the induction of MAA to be UV dependent peaking in the UV-B range at around 290 nm. The results indicate that the studied cyanobacterium, Anabaena sp. may protect itself from deleterious short wavelength solar radiation by its ability to synthesize a mycosporine-like amino acid in response to UV-B radiation and thereby screen the negative effects of UV-B.     

The role of UV-B radiation in aquatic and terrestrial ecosystems--an experimental and functional analysis of the evolution of UV-absorbing compounds

We analysed and compared the functioning of UV-B screening pigments in plants from marine, fresh water and terrestrial ecosystems, along the evolutionary line of cyanobacteria, unicellular algae, primitive multicellular algae, charophycean algae, lichens, mosses and higher plants, including amphibious macrophytes. Lichens were also included in the study. We were interested in the following key aspects: (a) does the water column function effectively as an 'external UV-B filter'?; (b) do aquatic plants need less 'internal UV-B screening' than terrestrial plants?; (c) what role does UV screening play in protecting the various plant groups from UV-B damage, such as the formation of thymine dimers?; and (d) since early land 'plants' (such as the predecessors of present-day cyanobacteria, lichens and mosses) experienced higher UV-B fluxes than higher plants, which evolved later, are primitive aquatic and land organisms (cyanobacteria, algae, lichens, mosses) better adapted to present-day levels of UV-B than higher plants? Furthermore, polychromatic action spectra for the induction of UV screening pigments of aquatic organisms have been determined. This is relevant for translating 'physical' radiation measurements of solar UV-B into 'biological' and 'ecological' effects. From the action spectra, radiation amplification factors (RAFs) have been calculated. These action spectra allow us to determine any mitigating or antagonistic effects in the ecosystems and therefore qualify the damage prediction for the ecosystems under study. We summarize and discuss the main results based on three years of research of four European research groups. The central theme of the work was the investigation of the effectiveness of the various screening compounds from the different species studied in order to gain some perspective of the evolutionary adaptations from lower to higher plant forms. The induction of mycosporine-like amino acids (MAAs) was studied in the marine dinoflagellate Gyrodinium dorsum, the green algal species Prasiola stipitata and in the cyanobacterium Anabaena sp. While visible (400-700 nm) and long wavelength UV-A (315-400 nm) showed only a slight effect, MAAs were effectively induced by UV-B (280-315 nm). The growth of the lower land organisms studied, i.e. the lichens Cladina portentosa, Cladina foliacaea and Cladonia arbuscula, and the club moss Lycopodiumannotinum, was not significantly reduced when grown under elevated UV-B radiation (simulating 15% ozone depletion). The growth in length of the moss Tortula ruralis was reduced under elevated UV-B. Of the aquatic plants investigated the charophytes Chara aspera showed decreased longitudinal growth under elevated UV-B. In the 'aquatic higher plants' studied, Ceratophyllum demersum, Batrachium trichophyllum and Potamogeton alpinus, there was no such depressed growth with enhanced UV-B. In Chara aspera, neither MAAs nor flavonoids could be detected. Of the terrestrial higher plants studied, Fagopyrum esculentum, Deschampsia antarctica, Vicia faba, Calamagrostis epigejos and Carex arenaria, the growth of the first species was depressed with enhanced UV-B, in the second species length growth was decreased, but the shoot number was increased, and in the latter two species of a dune grassland there was no reduced growth with enhanced UV-B. In the dune grassland species studied outdoors, at least five different flavonoids appeared in shoot tissue. Some of the flavonoids in the monocot species, which were identified and quantified with HPLC, included orientin, luteolin, tricin and apigenin. A greenhouse study with Vicia faba showed that two flavonoids (aglycones) respond particularly to enhanced UV-B. Of these, quercetin is UV-B inducible and mainly located in epidermal cells, while kaempferol occurs constitutively. In addition to its UV-screening function, quercetin may also act as an antioxidant. Polychromatic action spectra were determined for induction of the UV-absorbing pigments in three photosynthetic organisms, representing very different taxonomic groups and different habitats. In ultraviolet photobiology, action spectra mainly serve two purposes: (1) identification of the molecular species involved in light absorption; and (2) calculation of radiation amplification factors for assessing the effect of ozone depletion. Radiation amplification factors (RAFs) were calculated from the action spectra. In a somewhat simplified way, RAF can be defined as the percent increase of radiation damage for a 1% depletion of the ozone layer. Central European summer conditions were used in the calculations, but it has been shown that RAF values are not critically dependent on latitude or season. If only the ultraviolet spectral region is considered, the RAF values obtained are 0.7 for the green alga Prasiola stipitata, 0.4 for the dinoflagellate Gyrodinium dorsum, and 1.0 for the cyanobacterium Anabaena sp. In the case of P. stipitata, however, the effect of visible light (PAR, photosynthetically active radiation, 400-700 nm) is sufficient to lower the RAF to about 0.4, while the PAR effect for G. dorsum is negligible. RAFs for some damage processes, such as for DNA damage (RAF=2.1 if protective effects or photorepair are not considered [1]), are higher than those above. Our interpretation of this is that if the ozone layer is depleted, increased damaging radiation could overrule increased synthesis of protective pigments. In addition to investigating the functional effectiveness of the different screening compounds, direct UV effects on a number of key processes were also studied in order to gain further insight into the ability of the organisms to withstand enhanced UV-B radiation. To this end, the temperature-dependent repair of cyclobutane dimers (CPD) and (6-4) photoproducts induced by enhanced UV-B was studied in Nicotiana tabacum, and the UV-B induction of CPD was studied in the lichen Cladonia arbuscula. Also, photosynthesis and motility were monitored and the response related to the potential function of the screening compounds of the specific organism.     

Occurrence of Mycosporine-like Amino Acids (MAAs) from the Bloom-Forming Cyanobacteria Aphanizomenon Strains

Mycosporine-like amino acids (MAAs) are widespread in various microbes and protect them against harsh environments. Here, four different Aphanizomenon species were isolated from severely eutrophic waterbodies, Lake Dianchi and the Guanqiao fishpond. Morphological characters and molecular phylogenetic analysis verified that the CHAB5919, 5921, and 5926 strains belonged to the Aphanizomenon flos-aquae clade while Guanqiao01 belonged to the Aphanizomenon gracile clade. Full wavelength scanning proved that there was obvious maximal absorption at 334 nm through purified methanol extraction, and these substances were further analyzed by HPLC and UPLC-MS-MS. The results showed that two kinds of MAAs were discovered in the cultured Aphanizomenon strains. One molecular weight was 333.28 and the other was 347.25, and the daughter fragment patterns were in accordance with the previously articles reported shinorine and porphyra-334 ion characters. The concentration of the MAAs was calibrated from semi-prepared MAAs standards from dry cells of Microcystis aeruginosa PCC7806 algal powder, and the purity of shinorine and porphyra-334 were 90.2% and 85.4%, respectively. The average concentrations of shinorine and porphyra-334 were 0.307–0.385 µg/mg and 0.111–0.136 µg/mg in Aphanizomenon flos-aquae species, respectively. And there was only one kind of MAAs (shinorine) in Aphanizomenon gracile species.,with a content of 0.003–0.049 µg/mg dry weight among all Aphanizomenon gracile strains. The shinorine concentration in Aphanizomenon flos-aquae was higher than that in Aphanizomenon gracile strains. The total MAAs production can be ranked as Aphanizomenon flos-aquae > Aphanizomenon gracile.     

Synthesis of aminobismethylenephosphonic acids on a platform of <Emphasis Type="Italic">p-tert</Emphasis>-Butylthiacalix[4]arene in 1,3-<Emphasis Type="Italic">alternate</Emphasis> configuration

Aminobismethylenephosphonic acids on a platform of p-tert-butylthiacalix[4]arene were obtained by reacting macrocyclic amines with phosphorous acid and formaldehyde under acid catalysis. Free phenol hydroxy groups on the lower rim of p-tert-butylthiacalix[4]arene were found to inhibit the interaction with the amino moieties of the macrocycle. In the case of amino derivatives of thiacalix[4]arene containing no hydroxy groups the reaction led to the formation of target compounds in good yields.     

Dicarboxylic acids and hydroxy fatty acids in different species of fungi

Secondary metabolites of fungi can be responsible for allergies; therefore, the identification of compounds produced by these organisms is very important. Fungi produce large amounts of secondary metabolites, which belong to groups of chemicals such as: dicarboxylic acids, hydroxy acids, alcohols, hydrocarbons, esters, fatty acids, sterols, amino acids and mycotoxins. The presence of all these compounds in human proximity contributes to many diseases. Therefore, the aim of the study was a qualitative and quantitative analysis of hydroxy and dicarboxylic acids produced by fungi occurring in student hostel in Poland, in the province of Pomerania. The following species of fungi were subjected to extraction: Aspergillus niger, Aspergillus fumigatus, Aspergillus candidus, Rhizopus sp., Geotrichum candidum, and Penicillium chrysogenum. A mixture of ethyl acetate and methanol was used for the extraction. The obtained extracts were further analyzed by gas chromatography mass spectrometry (GC–MS). In all samples of fungi, the presence of a total of 22 acids, including 13 dicarboxylic and 9 hydroxy acids, was confirmed. Most acids (17 different acids) were identified in A. fumigatus. Only 10 acids were identified in the mycelium of G. candidum and A. niger. Acids which were identified in all samples of the mycelium were 22-hydroxydocosanoic acid, 24-hydroxytetracosanoic acid and adipic acid. The most abundant compounds were 22-hydroxydocosanoic acid in A. fumigatus, A. candidus, Rhizopus sp., G. candidum and P. chrysogenum, and succinic acid in A. niger. More experiments are needed to understand the physiological role of hydroxy and dicarboxylic acids. We hope that our results are an important contribution to further studies on the human health.     

Mycosporine-like amino acids in the marine dinoflagellate Gyrodinium dorsum: induction by ultraviolet irradiation

Cultures of the marine dinoflagellate Gyrodinium dorsum have been exposed to polychromatic radiation (photosynthetically active radiation and UV) from a solar simulator for up to 72 h. Different irradiance spectra in the ultraviolet are produced by inserting cut-off filters between lamp and samples. The mycosporine-like amino acid (MAA) content and composition are investigated by spectroscopic and chromatographic analysis. The study reveals that G. dorsum contains a complex mixture of several aminocyclohexenimine-MAAs and one aminocyclohexenone-MAA. UV irradiation around 320 nm induces an increase in the concentration of all MAAs in the samples. In contrast, exposure to short-wavelength UV-B radiation results in decreased overall MAA production. Furthermore, there is a spectral shift in the absorption of the MAA mixture towards shorter wavelengths, indicating that short-wavelength UV-B induces an altered MAA composition. The amount of MAAs is normalized to the chlorophyll a concentration.     

Photoprotection and long-term acclimation to UV radiation in the marine diatom Thalassiosira weissflogii.

Unialgal cultures of the marine diatom Thalassiosira weissflogii (Grunow) were exposed for 40 days to artificial UV-B radiation in the presence of PAR and UV-A to examine long-term acclimation to UV, PAR and UV-A were supplied 14 h daily, while UV-B (two levels: 0.16 and 0.30 W m(-2) unweighted) was supplied for 4 h/day. Growth rates and photochemical capacity (CFC ratio) both decreased over the first 10-15 days, then recovered. No obvious differences were noted between the responses to the two UV-B treatments. The concentration of the major pigments (chlorophyll a and c(1+2), fucoxanthin and beta,beta-carotene) changed very little with time, except for diatoxanthin. which increased over the first 16 days, decreased over the next 13 days, then increased again from day 29 to the end of the experiment. The concentration of total mycosporine-like amino acids (MAAs) was initially undetectable, then increased from day 16 in the high UV-B treatment and after day 22 in the low UV-B treatment, reaching a maximum on day 29 for both treatments and decreasing afterwards. The synthesis of MAAs proceeded only once photochemical capacity had recovered from the initial UV stress and this recovery likely involved the xanthophyll cycle (diatoxanthin increase). The concentration of MAAs decreased when the cells showed signs of photoinhibition (decrease in CFC ratio). It also showed an inverse trend with diatoxanthin. UV-B alone had little regulatory effect over these responses, except possibly for an earlier synthesis of MAAs under HUV-B conditions. This suggests that the observed changes were due to UV-A rather than to UV-B exposure. The overall response of this coastal diatom to prolonged UV exposure indicates that T. weissflogii is a relatively UV-tolerant species and that its long-term response to UV exposure involves an activation of the xanthophyll cycle followed by the synthesis of MAAs, which may proceed only when photoinhibition is relieved.     

Mycosporine-like Amino Acids Provide Protection Against Ultraviolet Radiation in Eggs of the Green Sea Urchin Strongylocentrotus droebachiensis

A photoprotective role of ultraviolet radiation-absorbing mycosporine-like amino acids (MAAs) in eggs of the green sea urchin Strongylocentrotus droebachiensis was demonstrated by comparing UV-induced delays in the first division of embryos having either high or low concentrations of MAAs. Embryos from adult urchins fed Laminaria saccharina (no MAAs) had low concentrations of MAAs and experienced a significantly longer UV-induced delay in cleavage (25.1%) than MAA-rich embryos from adults fed Mastocarpus stellatus (12.8% delay) or a combination diet of both macroalgae (12.3% delay). Collectively, these embryos displayed a significant inverse logarithmic relationship between MAA concentration and percentage cleavage delay, so that the greater the MAA concentration in the eggs, the less they were affected by UV radiation. This is the first study to examine such MAA manipulation of cellular MAA concentrations with no prior UV exposure of the experimental subjects. Concentrations of MAAs were also measured in unfertilized eggs, blastulae, gastrulae and early pluteus larvae, providing the first documentation of changes in MAAs during embryological and larval development. The concentration of shinorine (the principal MAA in the eggs) did not change during short-term UV exposure in vivo or long-term exposure in vitro; such photostability is a useful attribute of a natural sunscreen.     

Sulfur Deficiency Changes Mycosporine-like Amino Acid (MAA) Composition of Anabaena variabilis PCC 7937: A Possible Role of Sulfur in MAA Bioconversion

In the present investigation we show for the first time that bioconversion of a primary mycosporine-like amino acid (MAA) into a secondary MAA is regulated by sulfur deficiency in the cyanobacterium Anabaena variabilis PCC 7937. This cyanobacterium synthesizes the primary MAA shinorine (RT = 2.2 min, λ_max = 334 nm) under normal conditions (PAR + UV-A + UV-B); however, under sulfur deficiency, a secondary MAA palythine-serine (RT = 3.9 min, λ_max = 320 nm) appears. Addition of methionine to sulfur-deficient cultures resulted in the disappearance of palythine-serine, suggesting the role of primary MAAs under sulfur deficiency in recycling of methionine by donating the methyl group from the glycine subunit of shinorine to tetrahydrofolate to regenerate the methionine from homocysteine. This is also the first report for the synthesis of palythine-serine by cyanobacteria which has so far been reported only from corals. Addition of methionine also affected the conversion of mycosporine-glycine into shinorine, consequently, resulted in the appearance of mycosporine-glycine (RT = 3.6 min, λ_max = 310 nm). Our results also suggest that palythine-serine is synthesized from shinorine. Based on these results we propose that glycine decarboxylase is the potential enzyme that catalyzes the bioconversion of shinorine to palythine-serine by decarboxylation and demethylation of the glycine unit of shinorine.     

Photodegradation and photosensitization of mycosporine-like amino acids

The photodegradation and photosensitization of several mycosporine-like amino acids (MAAs) were investigated. The photodegradation of the MAA, palythine, was tested with three photosensitizers: riboflavin, rose bengal and natural seawater. For comparison of degradation rates, the riboflavin-mediated photosensitization of six other MAAs was also examined. When riboflavin was used as a photosensitizer in distilled water, MAAs were undetectable after 1.5h. Palythine showed little photodegradation when rose bengal was added as the photosensitizer (k=0.12x10(-3)m(2)kJ(-1)). Palythine dissolved in natural seawater containing high nitrate concentrations also showed slow photodegradation rate constants (k=0.26x10(-3)m(2)kJ(-1)) over a 24-h period of constant irradiation. Similar experiments in deep seawater with porphyra-334 and shinorine resulted in 75% of the initial MAA remaining after 4h of irradiation and rates of 0.018 and 0.026x10(-3) m(2) kJ(-1), respectively. Experiments conducted in deep seawater with riboflavin additions resulted in photodegradation rate constants between 0.77x10(-3) and 1.19x10(-3)m(2)kJ(-1) for shinorine and porphyra-334, respectively. Photoproduct formation appeared to be minimal with the presence of a dehydration product of the cycloheximine ring structure indicated as well as the presence of amino acids. Evidence continues to build for the role of MAAs as potent and stable UV absorbers. This study further highlights the photostability of several MAAs in both distilled and seawater in the presence of photosensitizers.     

Fatty acids and amino acids of entomopathogenic fungus <Emphasis Type="Italic">Conidiobolus coronatus</Emphasis> grow on minimal and rich media

Entomopathogenic fungi are referred to as potential candidates as insect pest control agents. The objective of the study was to identify fatty acids and amino acids from Conidiobolus coronatus cultured on two different media. Each medium was extracted with ethyl acetate and its mixtures with isopropanol, acetonitrile and methanol. Analyses of fatty acids and amino acids of entomopathogenic fungus C. coronatus were performed by means of gas chromatography coupled with mass spectrometry. The analysis showed that the fungus C. coronatus produces the following groups of compounds: fatty acids and amino acids; α- and β-glucopyranose were also identified. The identified fatty acids included 12–20, 22 and 24 carbon atoms per chain. The highest content of fatty acids was detected in a mycelium sample cultured in a liquid minimal medium extracted with ethyl acetate. The lowest content of these organic compounds was identified in mycelium cultured in a liquid nutrient-rich medium extracted with ethyl acetate–methanol mixture. Fatty acids were found to account for 62.0 mass % to 94.4 mass % of all organic compounds in the analyzed mycelia. C18:1 acids were detected in the highest amounts when ethyl acetate was used as the extracting agent. The identified amino acids accounted for 4 mass % to 21 mass % of all organic compounds. Upon extraction of C. coronatus mycelium samples with the ethyl acetate—methanol mixture, two anomeric forms of glucose were also identified. An analysis of the studied material confirmed, that the entomopathogenic fungus C. coronatus is a very rich source of organic compounds, which might encourage its further research so as to identify an even larger number of compounds being produced by this species.     

Physiological responses of Acropora cervicornis to increased solar irradiance

The effects of increased UV radiation (UV-B [280-320 nm] + UV-A [320-400 nm]; hereafter UVR) on the growth, production of photosynthetic pigments and photoprotective mycosporine-like amino acids (MAAs) were studied in the threatened Caribbean coral Acropora cervicornis transplanted from 20 to 1 m depth in La Parguera, Puerto Rico. The UVR exposure by the transplanted colonies was significantly higher than that at 20 m, while photosynthetically active radiation (PAR) only increased by 9%. Photosynthetic pigments, quantified with HPLC, as well as linear extension rates and skeletal densities, were significantly reduced 1 month after transplantation to 1 m depth, while MAAs increased significantly despite immediate paling experienced by transplanted colonies. While these colonies showed a significant reduction in photosynthetic pigments, there were no significant reductions in zooxanthellae densities suggesting photoacclimation of the coral's symbionts to the new radiation conditions. The results suggest that while corals might be able to survive sudden increases in UVR and PAR, their skeletal structure can be greatly debilitated due to a reduction in the photosynthetic capacity of their symbionts and a possible relocation of resources.     

Effect of Ultraviolet Radiation on Acetylcholinesterase Activity in Freshwater Copepods

We analyzed the effects of UV radiation (UVR) effects on acetylcholinesterase (AChE) activity in two calanoid copepods, Boeckella gibbosa and Parabroteas sarsi that inhabit Patagonian shallow lakes. We studied the effect of experimental UVR (UV-B and UV-A) exposure on AChE activity in relation to basal antioxidant capacities of both copepods. Our experiments showed that UVR can effectively depress AChE activity, although with differences between species. In both copepods AChE was affected by UV-B, whereas UV-A only affected AChE in B. gibbosa. Both copepods also differed in body elemental composition (C:N:P), photoprotecting compound content (carotenoids and mycosporine-like amino acids) and enzymatic antioxidant capacity (glutathione S-transferase [GST]). Our results suggest that when exposed to UVR, AChE activity would depend more on the antioxidant capacity (GST) and P availability for enzyme synthesis than on the photoprotective compounds.