Wavelength-dependent induction of UV-absorbing mycosporine-like amino acids in the red alga Chondrus crispus under natural solar radiation

Polychromatic response spectra for the induction of UV absorbing mycosporine-like amino acids (MAAs) were calculated after exposing small thalli of the red alga Chondrus crispus under various cut-off filters to natural solar radiation on the North Sea island Helgoland, Germany. The laboratory-grown specimens typically contain only traces of palythine and synthesise five different MAAs rapidly and in high concentrations after being transplanted into shallow water. The resulting qualitative and quantitative patterns of MAA induction differed markedly with respect to spectral distribution. Furthermore, the wavebands effective for MAA induction vary within the MAA. UV-B radiation had a negative effect on the accumulation of the major MAAs shinorine (λ_max=334 nm) and palythine (λ_max=320 nm), while short wavelength UV-A exhibits the highest quantum efficiency on their synthesis. In contrast, the synthesis of asterina-330 (λ_max=330 nm), palythinol (λ_max=332 nm) and palythene (λ_max=360 nm) was mainly induced by UV-B radiation. Whether the synthesis of shinorine and palythine is induced by a photoreceptor with an absorption maximum in the short wavelength UV-A and whether a second photoreceptor absorbing UV-B radiation is responsible for the induction of asterina-330, palythinol and palythene remains to be studied.Our results show that C. crispus has a high capacity to adapt flexibly the qualitative and quantitative MAA concentration to the prevailing spectral distribution of irradiance. On one hand, this is regarded as an important aspect with respect to the acclimation of algae to increasing UV-B irradiance in the context of ongoing depletion of stratospheric ozone. On the other hand, the experiment demonstrates that UV-A irradiance is more important for the induction of the major MAAs shinorine and palythine than UV-B.     

Mycosporine-like amino acids (MAAs) profile of a rice-field cyanobacterium Anabaena doliolum as influenced by PAR and UVR

The mycosporine-like amino acid (MAA) profile of a rice-field cyanobacterium, Anabaena doliolum, was studied under PAR and PAR + UVR conditions. The high-performance liquid chromatographic analysis of water-soluble compounds reveals the biosynthesis of three MAAs, mycosporine-glycine (lambda (max) = 310 nm), porphyra-334 (lambda (max) = 334 nm) and shinorine (lambda (max) = 334 nm), with retention times of 4.1, 3.5 and 2.3 min, respectively. This is the first report for the occurrence of mycosporine-glycine and porphyra-334 in addition to shinorine in Anabaena strains studied so far. The results indicate that mycosporine-glycine (monosubstituted) acts as a precursor for the biosynthesis of the bisubstituted MAAs shinorine and porphyra-334. Mycosporine-glycine was under constitutive control while porphyra-334 and shinorine were induced by UV-B radiation, indicating the involvement of UV-regulated enzymes in the biotransformation of MAAs. It seems that A. doliolum is able to protect its cell machinery from UVR by synthesizing a complex set of MAAs and thus is able to survive successfully during the summer in its natural brightly lit habitats.     

SYNTHESIS OF MYCOSPORINE-LIKE AMINO ACIDS IN CHONDRUS CRISPUS (FLORIDEOPHYCEAE) AND THE CONSEQUENCES FOR SENSITIVITY TO ULTRAVIOLET B RADIATION

The induction and protective role of the UV-absorbing compounds known as mycosporine-like amino acids (MAAs) were examined in sublittoral Chondrus crispus Stackh. transplanted for 2 weeks in the spring and summer to shallow water under three irradiance conditions: PAR (photosynthetically active radiation; 400–700 nm), PAR + UVA (PAR + 320– 400 nm), PAR + UVA + UVB (PAR + UVA + 280– 320 nm). Sublittoral thalli collected around Helgoland, North Sea, Germany, from 6 m below the mean low water of spring tides contained less than 0.1 mg·g⁻¹ dry weight (DW) total MAAs, whereas eulittoral samples contained over 1 mg·g⁻¹ DW. Transplantation to shallow water led to the immediate synthesis of three MAAs in the following temporal order: shinorine (λ_max 334 nm), asterina (λ_max 330 nm), and palythine (λ_max 320 nm), with the shinorine content peaking and then declining after 2 days (exposure to 100 mol photons·m⁻²). Maximum total MAA content (2 mg·g⁻¹ DW) also occurred after 2 days of induction, exceeding the content normally found in eulittoral samples. Furthermore, the relative proportion of the different MAAs at this time was different than that in eulittoral samples. After 2 days the total content declined to the eulittoral value, with palythine as the principal MAA. Similar data were obtained for all treatments, indicating that MAA synthesis in C. crispus was induced by PAR and not especially stimulated by UV radiation. The ability of photosystem II (PSII) to resist damage by UVB was tested periodically during the acclimation period by exposing samples to a defined UVB dose in the lab. Changes in chlorophyll fluorescence (F_v/F_m and effective quantum yield, φ_II) indicated that PSII function was inhibited during the initial stage of acclimation but gradually improved with time. No difference among screening treatments was detected except in spring for the samples acclimating to PAR + UVA + UVB. In this treatment F_v/F_m and φ_II were significantly lower than in the other treatments. During the first week of each experiment, growth rates were also significantly reduced by UVB. The reductions occurred despite maximum MAA content, indicating an incomplete protection of photosynthetic and growth-related processes.     

Discovery of Gene Cluster for Mycosporine-Like Amino Acid Biosynthesis from Actinomycetales Microorganisms and Production of a Novel Mycosporine-Like Amino Acid by Heterologous Expression

Mycosporines and mycosporine-like amino acids (MAAs), including shinorine (mycosporine-glycine-serine) and porphyra-334 (mycosporine-glycine-threonine), are UV-absorbing compounds produced by cyanobacteria, fungi, and marine micro- and macroalgae. These MAAs have the ability to protect these organisms from damage by environmental UV radiation. Although no reports have described the production of MAAs and the corresponding genes involved in MAA biosynthesis from Gram-positive bacteria to date, genome mining of the Gram-positive bacterial database revealed that two microorganisms belonging to the order Actinomycetales, Actinosynnema mirum DSM 43827 and Pseudonocardia sp. strain P1, possess a gene cluster homologous to the biosynthetic gene clusters identified from cyanobacteria. When the two strains were grown in liquid culture, Pseudonocardia sp. accumulated a very small amount of MAA-like compound in a medium-dependent manner, whereas A. mirum did not produce MAAs under any culture conditions, indicating that the biosynthetic gene cluster of A. mirum was in a cryptic state in this microorganism. In order to characterize these biosynthetic gene clusters, each biosynthetic gene cluster was heterologously expressed in an engineered host, Streptomyces avermitilis SUKA22. Since the resultant transformants carrying the entire biosynthetic gene cluster controlled by an alternative promoter produced mainly shinorine, this is the first confirmation of a biosynthetic gene cluster for MAA from Gram-positive bacteria. Furthermore, S. avermitilis SUKA22 transformants carrying the biosynthetic gene cluster for MAA of A. mirum accumulated not only shinorine and porphyra-334 but also a novel MAA. Structure elucidation revealed that the novel MAA is mycosporine-glycine-alanine, which substitutes l-alanine for the l-serine of shinorine.     

BLUE LIGHT AND UV-A RADIATION CONTROL THE SYNTHESIS OF MYCOSPORINE-LIKE AMINO ACIDS IN CHONDRUS CRISPUS (FLORIDEOPHYCEAE)

The induction of UV-absorbing compounds known as mycosporine-like amino acids (MAAs) by red, green, blue, and white light (43% ambient radiation greater than 390 nm) was examined in sublittoral Chondrus crispus Stackh. Fresh collections or long-term cultures of sublittoral thalli, collected from Helgoland, North Sea, Germany, and containing no measurable amounts of MAAs, were exposed to filtered natural radiation for up to 40 days. The MAA palythine (λ_max 320 nm) was synthesized in thalli in blue light to the same extent observed in control samples in white light. In contrast, thalli in green or red light contained only trace amounts of MAAs. After the growth and synthesis period, the photosynthetic performance of thalli in each treatment, measured as pulse amplitude modulated chlorophyll fluorescence, was assessed after a defined UV dose in the laboratory. Thalli with MAAs were more resistant to UV than those without, and exposure to UV-A+B was more damaging than UV-A in that optimal (F_v/F_m) and effective (φ_II) quantum yields were lower and a greater proportion of the primary electron acceptor of PSII, Q, became reduced at saturating irradiance. However, blue light-grown thalli were generally more sensitive than white light control samples to UV-A despite having similar amounts of MAAs. The most sensitive thalli were those grown in red light, which had significantly greater reductions in F_v/F_m and φ_II and greater Q reduction. Growth under UV radiation alone had been shown previously to lead to the synthesis of the MAA shinorine (λ_max 334 nm) rather than palythine. In further experiments, we found that preexposure to blue light followed by growth in natural UV-A led to a 7-fold increase in the synthesis of shinorine, compared with growth in UV-A or UV-A+B without blue light pretreatment. We hypothesize that there are two photoreceptors for MAA synthesis in C. crispus, one for blue light and one for UV-A, which can act synergistically. This system would predispose C. crispus to efficiently synthesize UV protective compounds when radiation levels are rising, for example, on a seasonal basis. However, because the UV-B increase associated with artificial ozone reduction will not be accompanied by an increase in blue light, this triggering mechanism will have little additional adaptive value in the face of global change unless a global UV-B increase positively affects water column clarity.     

Distribution of mycosporine-like amino acids in the sea hare Aplysia dactylomela: effect of diet on amounts and types sequestered over time in tissues and spawn

We investigated the interaction of diet and accumulation of UV-absorbing mycosporine-like amino acids (MAAs) in body tissues and spawn of the sea hare Aplysia dactylomela to determine if MAA accumulation reflects type and level of dietary intake. Food sources were the red algae Acanthophora spicifera, Centroceras clavulatum, and Laurencia sp., and the green alga, Ulva lactuca. Adults were maintained on these foods for 40 days, after which feces were collected and tissues separated by dissection. Field animals were similarly sampled at this time. All spawn from experimental and field animals was collected over the study period. Samples, including seaweed foods, were analysed for six MAAs. Overnight consumption experiments using a variety of common seaweeds and one seagrass from A. dactylomela's habitat showed that the four seaweeds selected as foods were among those best-eaten by Aplysia. After 40 days levels of specific MAAs in the tissues of experimental animals showed excellent correlation with those in their diets, suggesting that the MAAs were dietarily-derived. Relative MAA contents in spawn from all diet groups correlated well with those in spawn from field animals. Commonest MAAs in spawn were porphyra-334, shinorine, and palythine, in this order. Concentrations of these MAAs were maintained at constant levels over time in spawn from all diet groups eating red algae and from field animals. Spawn from the Ulva dietary group showed an initial significant decline in MAA concentrations, but levels stabilized after the first 2 weeks. Skin was rich in porphyra-334 and shinorine, and levels of these in experimental animals correlated well with comparable levels in the skin of field animals. Digestive glands contained high levels of asterina-330, particularly those of the Centroceras dietary group, where concentrations reached a maximum of 21 mg dry g(-1).     

Natural ultraviolet radiation and photosynthetically active radiation induce formation of mycosporine-like amino acids in the marine macroalga Chondrus crispus (Rhodophyta)

The UV-absorbing mycosporine-like amino acids (MAAs) are hypothesized to protect organisms against harmful UV radiation (UVR). Since the physiology and metabolism of these compounds are unknown, the induction and kinetics of MAA biosynthesis by various natural radiation conditions were investigated in the marine red alga Chondrus crispus collected from Helgoland, Germany. Three photosynthetically active radiation (PAR, 400–700 nm) treatments without UVR and three UV-A/B (290–400 nm) treatments without PAR were given. Chondrus crispus collected from 4–6 m depth contained only traces of the MAA palythine. After 24 h exposure to 100% ambient PAR, traces of three additional MAAs, shinorine, palythinol and palythene, were detected, and their concentrations increased strongly during a one-week exposure to all PAR treatments. The concentration of all MAAs varied directly with PAR dose, with palythine and shinorine being four- to sevenfold higher than palythinol and palythene. Likewise, naturally high doses of both UV-A and UV-B resulted in a strong accumulation of all MAAs, in particular shinorine. While shinorine accumulation was much more stimulated by UVR, the content of all other MAAs was more affected by high PAR, indicating an MAA-specific induction triggered by UVR or PAR. Received: 24 September 1997 / Accepted: 17 December 1997     

Ultraviolet and osmotic stresses induce and regulate the synthesis of mycosporines in the cyanobacterium Chlorogloeopsis PCC 6912

The cyanobacterium Chlorogloeopsis PCC 6912 was found to synthesize and accumulate two putative UV sunscreen compounds of the mycosporine (mycosporine-like amino acid; MAA) type: mycosporine-glycine and shinorine. These MAAs were not constitutively present in the cells; their synthesis could be induced specifically either by exposure to UVB radiation (280–320 nm) or by osmotic stress, but not by other stress factors such as heat or cold shock, nutrient limitation, or photooxidative stress. A significant synergistic enhancement of MAA synthesis was observed when both stress factors were applied in combination. Although osmotic stress could induce MAA synthesis, comparison of the intracellular contents of MAAs with those of sugar osmolytes (glucose and trehalose) indicated that MAAs play no significant role in attaining osmotic homeostasis. UVB strongly enhanced the accumulation of shinorine, whereas osmotic stress had a more pronounced effect on mycosporine-glycine. This differential effect on the steady-state contents of each MAA could be explained either by differential regulation of biosynthesis or by differential loss rates of MAAs (leakage) under each condition. A preferential leakage of mycosporine-glycine from the cells after a hypoosmotic shock was detected. The results are interpreted in terms of an adaptive necessity for a combined regulatory control responding to both UV and external osmotic conditions in organisms that accumulate water-soluble sunscreens intracellularly. Received: 26 March 1999 / Accepted: 13 July 1999     

Ultraviolet sunscreen compounds in epiphytic red algae from mangroves

Epiphytic red algae of the order Ceramiales from mangroves and salt marshes (nine species from Bostrychia, three from Stictosiphonia and four from Caloglossa) produce varying levels of the UV-absorbing compounds mycosporine-glycine, shinorine, porphyra-334, palythine, asterina-330 and palythinol, a suite of substances chemically assigned as mycosporine-like amino acids (MAAs). Mean MAA levels varied from 0.02 to 12.8 mg g^–1 DW in field-collected and laboratory cultured specimens. While in field samples of Bostrychia montagneiHarvey, Bostrychia radicans (Montagne) Montagne and Caloglossa apomeiotica J.West et G.Zuccarello MAA concentrations were generally higher compared to cultured plants of the same taxa, Bostrychia tenella(Lamouroux) J.Agardh did not show such a difference. Catenella caespitosa (Withering) L.Irvine, Catenella impudica (Montagne) J.Agardh and Catenella nipae Zanardini (Gigartinales, Caulacanthaceae) produce two novel UV-absorbing compounds: MAA-1 (1.4–4.3 mg g ^–1 DW) and MAA-2 (0.1–1.0 mg g^–1 DW), which absorb at 334 nm and 320 nm, respectively. In laboratory culture of Bostrychia moritziana when photosynthetically active radiation (PAR) was increased from 20 to 40 mol photons m^–2 s^–1, the total level of palythinol increased by 85% (from 2.0 to 3.7 mg g^–1 DW). In a culture of Caloglossa leprieurii when PAR was increased from 40 to 80 mol m^–2 s^–1the porphyra-334 content increased by 77% (from 3.1 to 5.5 mg g^–1 DW). Extremely high MAA contents of >30 mg g^–1 DW were detected in mature tetrasporangial sori prepared from two isolates of laboratory-cultured reproductive Caloglossa apomeiotica compared to vegetative plants (about 10 mg MAAs g^–1 DW) indicating tetraspores loaded up with UV-sunscreens. All data demonstrate that mangrove red algae contain high MAA concentrations, particularly the reproductive structures, and hence these compounds may act as biochemical photoprotectants against exposure to UV-radiation.     

The effect of ultraviolet radiation on photosynthesis and ultraviolet-absorbing substances in the endemic Arctic macroalga Devaleraea ramentacea (Rhodophyta)

In field studies conducted at the Kongsfjord (Spitsbergen), the effect of filtered natural radiation conditions (solar without ulraviolet [UV]-A+UV-B, solar without UV-B, solar) on photosynthesis and the metabolism of UV-absorbing mycosporine-like amino acids (MAAs) in the marine red alga Devaleraea ramentacea have been studied. While solar treatment without UV-A+UV-B did not affect photosynthesis during the course of a day, solar without UV-B and the full solar spectrum led to a strong inhibition. However, after offset of the various radiation conditions, all algae fully recovered. Isolates collected from different depths were exposed in the laboratory to artificial fluence rates of photosynthetic active radiation (PAR), PAR+UV-A, and PAR+UV-A+UV-B. The photosynthetic capacity was affected in accordance with the original sampling depth, i.e. shallow-water isolates were more resistant than algae from deeper waters, indicating that D. ramentacea is able to acclimate to changes in irradiance. Seven different UV-absorbing MAAs were detected in this alga, namely mycosporine-glycine, shinorine, porphyra-334, palythine, asterina-330, palythinol, and palythene. The total amount of MAAs continuously decreased with increasing collecting depth when sampled in mid June, and algae taken in late August from the same depths contained on average 30–45% higher MAA concentrations, indicating a seasonal effect as well. The presence of increasing MAA contents with decreasing depth correlated with a more insensitive photosynthetic capacity under both UV-A and UV-B treatments. Populations of D. ramentacea collected from 1 m depth, with one fully exposed to solar radiation and the other growing protected as understorey vegetation underneath the kelp Laminaria saccharina, exhibited quantitatively different MAA compositions in the apices. The exposed seaweeds contained 2.5-fold higher MAA values compared with the more shaded algae. Moreover, the exposed isolates showed a strong tissue gradient in MAAs, pigments, and proteins. The green apices contained 5-fold higher MAA contents than the red bases. Transplantation of D. ramentacea from 2 m depth to the surface induced the formation and accumulation of MAAs after 1 week exposure to the full solar spectrum. Control samples which were treated with the solar spectrum without UV-A+B or with solar without UV-B showed unchanged MAA contents, indicating a strong UV-B effect on MAA metabolism. All data well supported the suggested physiological function of MAAs as natural UV sunscreens in macroalgae.     

Heme/heme redox interaction and resolution of individual optical absorption spectra of the hemes in cytochrome bd from Escherichia coli

Cytochrome bd is a terminal component of the respiratory chain of Escherichia coli catalyzing reduction of molecular oxygen to water. It contains three hemes, b₅₅₈, b₅₉₅, and d. The detailed spectroelectrochemical redox titration and numerical modeling of the data reveal significant redox interaction between the low-spin heme b₅₅₈ and high-spin heme b₅₉₅, whereas the interaction between heme d and either hemes b appears to be rather weak. However, the presence of heme d itself decreases much larger interaction between the two hemes b. Fitting the titration data with a model where redox interaction between the hemes is explicitly included makes it possible to extract individual absorption spectra of all hemes. The α- and β-band reduced-minus-oxidized difference spectra agree with the data published earlier ([22] J.G. Koland, M.J. Miller, R.B. Gennis, Potentiometric analysis of the purified cytochrome d terminal oxidase complex from Escherichia coli, Biochemistry 23 (1984) 1051-1056., and [23] R.M. Lorence, J.G. Koland, R.B. Gennis, Coulometric and spectroscopic analysis of the purified cytochrome d complex of Escherichia coli: evidence for the identification of “cytochrome a₁” as cytochrome b₅₉₅, Biochemistry 25 (1986) 2314-2321.). The Soret band spectra show λ_max = 429.5 nm, λ_min ≈ 413 nm (heme b₅₅₈), λ_max = 439 nm, λ_min ≈ 400 ± 1 nm (heme b₅₉₅), and λ_max = 430 nm, λ_min = 405 nm (heme d). The spectral contribution of heme d to the complex Soret band is much smaller than those of either hemes b; the Soret/α (ΔA₄₃₀:ΔA₆₂₉) ratio for heme d is 1.6.     

Antioxidant activity of mycosporine-like amino acids isolated from three red macroalgae and one marine lichen

Several standard in vitro assays were performed in order to determine the potential antioxidant capabilities of purified aqueous extracts of the mycosporine-like amino acids (MAAs), porphyra-334 plus shinorine (P-334 + SH), isolated from the red alga Porphyra rosengurttii, asterina-330 plus palythine (AS-330 + PNE), from the red alga Gelidium corneum, shinorine (SH), from the red alga Ahnfeltiopsis devoniensis, and mycosporine -glycine (M-Gly), isolated from the marine lichen Lichina pygmaea. The scavenging potential of hydrosoluble radicals (ABTS⁺ decolorization method), the antioxidant activity in lipid medium (β-carotene/ linoleate bleaching method) and the scavenging capacity of superoxide radicals (pyrogallol autooxidation assay) were evaluated. In terms of scavenging of hydrosoluble radicals, the antioxidant activity of all MAAs studied was dose-dependent and it increased with the alkalinity of the medium (pH 6 to 8.5). M-Gly presented the highest activity in all pH tested; at pH 8.5 its IC₅₀ was 8-fold that of L-ascorbic acid (L-ASC) followed by AS-330 + PNE while P-334 + SH and SH showed scarce activity of scavenging of hydrosoluble free radicals. AS-330 + PNE showed high activity for inhibition of β-carotene oxidation relative to vitamin E and superoxide radical scavenging whilst the activity of P-334 +SH and SH were moderate. According to these results, the potential of MAAs in photoprotection can be considered high due to a double function: (1) UV chemical screening with high efficiency for UVB and UVA regions of the solar spectrum, and (2) their antioxidant capacity.     

Mycosporine-like amino acids in azooxanthellate and zooxanthellate early developmental stages of the soft coral Heteroxenia fuscescens

The ontogenetic changes of MAAs in the soft coral Heteroxenia fuscescens was studied in relation to their symbiotic state (azooxanthellate vs. zooxanthellate) under different temperature conditions in the Gulf of Eilat, northern Red Sea. The HPLC chromatograms for extracts of the planulae, azoo- and zooxanthellate primary polyps of H. fuscescens from all dates of collection yielded a single peak at 320 nm that has been identified as the compound palythine. Concentration of palythine in planulae at 23 °C was 7.57 ± 1 nmol mg^− 1 protein and at 28 °C reached 17.29 ± 1 nmol × mg^− 1 protein. Concentration of palythine in azooxanthellate primary polyps was 16.4 ± 3 nmol × mg^− 1 protein and 28.37 ± 2.8 nmol × mg^− 1 protein at 23 °C and 28 °C respectively. The palythine concentration for zooxanthellate primary polyps at 23 °C was 13 ± 3 nmol × mg^− 1 protein and at 28 °C 32.7 ± 2 nmol mg^− 1 protein. Palythine concentrations were significantly higher at 28 °C in the different animal groups and correlated linearly with the ambient collection temperature. This study shows for the first time that UVR and temperature act synergistically and affect the MAA levels of early life-history stages of soft corals.     

Mycosporine-Like Amino Acids Extracted from Scallop (<Emphasis Type="Italic">Patinopecten yessoensis</Emphasis>) Ovaries: UV Protection and Growth Stimulation Activities on Human Cells

Scallops (Patinopecten yessoensis) are extensively cultured and landed in Japan. During the processing of scallops, large amounts of internal organs and shells are discharged as industrial wastes. To reduce the burden on the environment, effective utilization and disposal methods of the wastes are required. Therefore, we have screened for useful materials in scallop internal organs, and found ultraviolet (UV) absorbing compounds from scallop ovaries. Based on UV absorption, electrospray ionization-mass spectrometry (ESI-MS), ESI-MS/MS, and nuclear magnetic resonance (NMR) spectra, three UV absorbing compounds were identified as mycosporine-like amino acids (MAAs): shinorine, porphyra-334 (P-334), and mycosporine-glycine. To investigate whether MAAs can act as a UV protector for human cells, we examined the protective effects of the three MAAs on human fibroblast cells from UV irradiation. All of the three examined MAAs protected the cells from UV-induced cell death. In particular, mycosporine-glycine had the strongest effect. Further, we found a promotion effect of MAAs on the proliferation of human skin fibroblast cells. From these results, it was found that the three MAAs isolated from scallop ovaries have a protective effect on human cells against UV light. MAAs have potential applications in cosmetics and toiletries as a UV protectors and activators of cell proliferation.     

Occurrence of mycosporine-like amino acids in the red-tide dinoflagellate Alexandrium excavatum: UV-photoprotective compounds?

Water extracts of the red-tide dinoflagellate Alexandrium excavatumgrown at ‘high’ light intensity (200 µE m^–2s^–1) show a broad absorbance maximum in the UV regionof the spectrum (310–360 nm). Using TLC and reverse-phaseHPLC a series of mycosporine-like amino acids have been characterized:mycosporine-glycine (_max = 310 nm), palythine (_max = 320 nm),asterina-330 (_max = 330 nm), shinorine (_max = 334 nm), porphyra-334(_max= 334 nm), palythenic acid (_max = 337 nm) and the isomericmixture of usujirene and palythene (_max = 359 nm). From theobserved spectral changes during transference from ‘low’(20 µE m^–2 s^–1) to ‘high’ (200µE m^–2 s^–1) light intensities and vice versa,the series of compounds are supposed to be biogenically relatedto one another. The presence of these compounds in A.excavatumis discussed in relation to their possible role in the photoprotectionto deleterious UV radiation.     

An ATP-grasp ligase involved in the last biosynthetic step of the iminomycosporine shinorine in Nostoc punctiforme ATCC 29133

We investigated the genetic basis for mycosporine sunscreen biosynthesis by the cyanobacterium Nostoc punctiforme ATCC 29133. Heterologous expression in Escherichia coli of three contiguous N. punctiforme genes (NpR5600, NpR5599, and NpR5598, here named mysA, mysB, and mysC, respectively) led to the production of mycosporine-glycine, an oxomycosporine. Additional expression of gene NpF5597 (mysD) led to the conversion of mycosporine-glycine into iminomycosporines (preferentially shinorine but also others like mycosporine-2-glycine and porphyra-334). This represents a new mode of enzymatic synthesis for iminomycosporines, one that differs in genetic origin, mechanism, and apparent substrate specificity from that known in Anabaena variabilis ATCC 29413. These results add to the emerging profile of the protein family of ATP-dependent ligases, to which the mysC product belongs, as important condensation enzymes in microbial secondary metabolism.     

Mycosporine-like amino acids in planktonic organisms living under different UV exposure conditions in Patagonian lakes

Mycosporine-like amino acids (MAAs) were studied in zooplanktonfrom 13 Argentinian lakes covering a broad range in altitude,maximum depth and physico-chemical properties of the water.Four to nine different MAAs (predominantly porphyra-334 andshinorine) were found in the copepods Boeckella gibbosa, B.gracilipes, B. meteoris and Parabroteas sarsi, and in the ciliateStentor amethystinus, while MAAs were undetectable in the cladoceranDaphnia middendorffiana. Among the different copepods, maximumMAA concentrations accounted for 0.25–1.31% of the dryweight, and contents were generally about three to seven times(up to 43 times) higher in the animals living in the clearestlakes compared to those occurring in low-UV systems. This variabilityin the content of MAAs was related to the lake altitude (r²= 0.71), and the fraction of the water column to which 1% ofthe surface UV radiation at 320 nm penetrated (r² = 0.57). Ourdata therefore underscore the role of MAAs as sunscreens todecrease the potential negative effects of solar radiation,but they also indicate that other environmental factors besidesUV transparency play a role in determining MAA concentrations.One lake was selected to obtain additional information on thequalitative composition of MAAs in seston of <100 µmbetween two sampling sites and over a 2 month study period (australsummer). Six different MAAs were detected in the samples, withporphyra-334 and palythine being predominant. In the copepodscollected simultaneously, there was low variation in MAA concentrationsbetween the two sites and over time. Thus, our results suggestthat under similar UV exposure conditions MAA contents of planktonicorganisms show low temporal variation.     

Temporal concentrations of sunscreen compounds (Mycosporine-like Amino Acids) in phytoplankton and in the New Zealand krill, Nyctiphanes australis G.O. Sars

This study investigated the relationship between seasonal changesin ambient UV-R, and sunscreen concentrations in phytoplanktonand krill. Concentrations of mycosprine-like amino acid (MAA)sunscreens were quantified in phytoplankton communities andin the krill Nyctiphanes australis over a 1-year period offthe Otago Coast, New Zealand. Ambient UV-B and UV-A ranged froma minimum mean daily dose of 2.19 x 10⁴ kJ day^–1 and 0.73x 10⁶ kJ day^–1 in June, to a maximum in January of 20.19x 10⁴ kJ day^–1 and 4.88 x 10⁶ kJ day^–1, respectively.Concentrations of MAAs (consisting almost entirely of Mycosporine-glycine)in the phytoplankton community were lowest in August (5.6 nmolµg^–1 Chl) when UV-R irradiances were minimal andhighest in January (41.4 nmol µg^–1 Chl) when UV-Rirradiances were maximal. Nyctiphanes australis was found tocontain five identified MAAs (mycosporine-glycine, shinorine,Porphyra-334, palythine and palythinol) and several unknownUV-R absorbing compounds. Concentrations ranged from 4.73 to15.51 nmol mg^–1 dw, with little indication of a seasonalcycle that could be correlated with changes in either phytoplanktonMAA concentrations or ambient UV-R irradiances. The findingssuggest that krill are neither accumulating MAAs in responseto changes in MAA concentrations in their phytoplankton food,or that MAA concentrations in krill are increased in responseto higher ambient UV-R irradiances. Concentrations of MAAs inkrill body parts (carapace, legs, eyes, antennae, muscle) weresimilar (4.89–5.98 nmol mg^–1 dw), with the exceptionof the carapace (2.03 nmol mg^–1 dw).     

Enzymatic and non-enzymatic defense mechanisms against ultraviolet-B radiation in two Anabaena species

Enzymatic and non-enzymatic defense strategies against ultraviolet-B radiation (UV-B, 280–315 nm) were studied in Anabaena doliolum and Anabaena strain L31, two of the most common strains of Indian cyanobacteria. Upon UV-B irradiation, both strains showed a 2–5-fold increase in antioxidative enzymes, such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD), as compared to non-irradiated control cultures. These enzymes scavenge damaging reactive oxygen species (ROS), generated by UV-B radiation inside the cells. In addition, these organisms also synthesize mycosporine-like amino acids (MAAs) which are able to carry out UV-screening and/at the same time as UV-quenching. The identification and characterization of three types of MAAs from both Anabaena species were performed using absorption spectroscopy, high performance liquid chromatography (HPLC), electro spray ionization-mass spectrometry (ESI-MS), Fourier Transform Infrared (FTIR) spectroscopy and Nuclear Magnetic Resonance (NMR) spectroscopy. Shinorine was found to be the most common MAA in both Anabaena species while porphyra-334 and mycosporine-glycine were present only in A. doliolum. The results of the present investigation clearly demonstrate that both enzymatic and non-enzymatic defense mechanisms are being employed by A. doliolum and Anabaena strain L31 to counteract the damaging effects of UV-B radiation.