UV-B-induced synthesis of photoprotective pigments and extracellular polysaccharides in the terrestrial cyanobacterium Nostoc commune.

Liquid cultures of the terrestrial cyanobacterium Nostoc commune derived from field material were treated with artificial UV-B and UV-A irradiation. We studied the induction of various pigments which are though to provide protection against damaging UV-B irradiation. First, UV-B irradiation induced an increase in carotenoids, especially echinenone and myxoxanthophyll, but did not influence production of chlorophyll a. Second, an increase of an extracellular, water-soluble UV-A/B-absorbing mycosporine occurred, which was associated with extracellular glycan synthesis. Finally, synthesis of scytonemin, a lipid-soluble, extracellular pigment known to function as a UV-A sunscreen, was observed. After long-time exposure, the UV-B effect on carotenoid and scytonemin synthesis ceased whereas the mycosporine content remained constantly high. The UV-B sunscreen mycosporine is exclusively induced by UV-B (< 315 nm). The UV-A sunscreen scytonemin is induced only slightly by UV-B (< 315 nm), very strongly by near UV-A (350 to 400 nm), and not at all by far UV-A (320 to 350 nm). These results may indicate that the syntheses of these UV sunscreens are triggered by different UV photoreceptors.     

Antioxidative activity and chemical constituents of edible terrestrial alga Nostoc commune Vauch

The extract of terrestrial alga Nostoc commune Vauch. has high antioxidative activity. Our study on N. commune Vauch. resulted in the isolation of two β-ionone derivatives, nostocionone and 3-oxo-β-ionone, together with four indole alkaloids, scytonemin, reduced scytonemin, N-(p-coumaroyl)tryptamine, and N-acetyltryptamine. The structures of the isolated compounds were determined on the basis of 1D and 2D NMR and MS analyses. Among these isolates, nostocionone and reduced scytonemin demonstrated strong antioxidative activities which were assessed by using a β-carotene oxidation assay.     

ROLE OF ULTRAVIOLET RADIATION IN MAINTAINING THE THREE-DIMENSIONAL STRUCTURE OF A CYANOBACTERIAL MAT COMMUNITY AND FACILITATING NITROGEN FIXATION

Cyanobacterial mat communities were collected in the mangrove forest bordering the Grand Cul de Sac Marin, Guadeloupe, French West Indies, which supports a community of nitrogen fixing cyanobacterial mats established on the trunk and branches of black mangrove ( Avicennia germinans L.). This study presents results that are focused on the mat community and the physiological and morphological adaptations to UV radiation. The dominant surface species of the mat, Nostoc cf commune Vaucher and Scytonema sp., possessed the UV-shielding pigment scytonemin. Mats grown on medium D agar without nitrogen under photosynthetically active radiation (PAR) only, rapidly became disorganized compared with those exposed to PAR + UV-A (320– 400 nm) + UV-B (280–320 nm) irradiation. Concurrent with disorganization, acetylene reduction activity (ARA = one third of N₂ reduction) was severely reduced, whereas mats irradiated with PAR + UV-A + UV-B maintained high ARA activity. Mats incubated for 27 days under PAR + UV-A + UV-B then exposed to PAR only exhibited a 68% stimulation of ARA, whereas ARA values were 33% inhibited in mats incubated with PAR only and then exposed to PAR + UV-A + UV-B. This favorable equilibrium was facilitated by the mats' three-dimensional structure in which the most UV-resistant species, N. commune , covers the surface with UV-sensitive species below this protective covering. The UV stressor was essential for the maintenance of mat structure and ARA.     

A comparison of scytonemin and its carbon analogue in terms of antioxidant properties through free radical mechanisms and conformational analysis: a DFT investigation

Scytonemin is a UV absorbing sheath pigment synthesized uniquely by cyanobacteria. Its biological features has attracted interest ecologically (in microbial mat systems), medically (for therapeutic activity) and astrobiologically (as a key biomarker). Recently, a carbon analogue of scytonemin, in which two nitrogen atoms are replaced by carbon atoms was synthesized to elucidate the origin of biological activity by comparison with scytonemin. In this work, their structural/conformational aspects and relative antioxidant capacity are compared making use of DFT calculations to provide insight about the similarities and differences between the two. The carbon analogue of scytonemin, isoelectronic with scytonemin, has the same structural skeleton and a similar potential energy surface but the hydrogens on the carbons that replace the nitrogens cause the phenolic rings to rotate out of the plane which is obseved for scytonemin. Thermochemically, the carbon analogue of scytonemin prefers the same radical scavenging mechanism scytonemin does, the HAT mechanism, and has a lower homolytic bond dissociation enthalpy for the OH group than that of scytonemin and other known antioxidants like ascorbic acid. The carbon analogue of scytonemin is suggested to be a novel synthetic antioxidant.     

Effects of nitrogen source on the synthesis of the UV-screening compound, scytonemin, in the cyanobacterium Nostoc punctiforme PCC 73102

The effects of nitrogen source (N(2), NO(3)(-) and NH(4)(+)) on scytonemin synthesis were investigated in the heterocystous cyanobacterium Nostoc punctiforme PCC 73102. With the required UVA radiation included, Nostoc synthesized three to seven times more scytonemin while fixing nitrogen than when utilizing nitrate or ammonium. A similar increase in scytonemin synthesis occurred when nitrate or ammonium became depleted by growth and Nostoc switched to diazotrophic metabolism with the differentiation of heterocysts. In addition, UVA-exposed cultures grown in medium with both NO(3)(-) and NH(4)(+) synthesized some scytonemin but synthesis increased when NH(4)(+) was depleted and growth had become dependent on NO(3)(-) reduction. Although the mechanism is unclear, these results suggest that the greater the restriction in nitrogen accessibility, the greater the production of scytonemin. Perhaps the entire response may be an interaction between this restriction and a resultant sensitivity to UV radiation that acts as a cue for determining the level of scytonemin synthesis. Scytonemin is a stable UVR screening compound and appears to be synthesized by cyanobacteria as a long-term solution for reducing UVR exposure and damage, but mainly or solely, when metabolic activity is absent. It is likely that during metabolic resurgence, the presence of a dense scytonemin sheath would facilitate the recovery process without the need for active defenses against UV radiation.     

UV-acclimation responses in natural populations of cyanobacteria (Calothrix sp.)

Phenotypic acclimation to changing conditions is typically thought to be beneficial to organisms in the environment. UV radiation is an important parameter affecting photosynthetic organisms in natural environments. We measured the response of photosynthetic carbon fixation in populations of cyanobacteria inhabiting a hot spring following acclimation to different UV treatments. These two very closely related populations of cyanobacteria, differing in their content of the extracellular UV-screening pigment scytonemin, were acclimated in situ under natural solar irradiance modified by filters that excluded both UVA/B, only UVB or transmitted both UVA/B. Cells from each preacclimation treatment were subsequently assayed for photosynthetic performance under all UV conditions (incubation treatment) giving a two-factor experimental design for each population. No acclimation filter treatment effects were observed even after two months under different acclimation treatments. This suggests that UV photoacclimation does not occur in either of these populations, regardless of the presence of scytonemin. By contrast, cells showed significant UV-inhibition during 1 h incubations under full sun. The population with high levels of scytonemin usually had lower rates of photosynthetic carbon fixation than the scytonemin-lacking population. However, the degree of UV inhibition, especially UVA inhibition, was higher for the cells without scytonemin pigment. These results suggest that closely related natural cyanobacterial populations respond differently to natural irradiance conditions and may be adopting different strategies of UV tolerance.     

The response regulator Npun_F1278 is essential for scytonemin biosynthesis in the cyanobacterium Nostoc punctiforme ATCC 29133

Following exposure to long‐wavelength ultraviolet radiation (UVA), some cyanobacteria produce the indole‐alkaloid sunscreen scytonemin. The genomic region associated with scytonemin biosynthesis in the cyanobacterium Nostoc punctiforme includes 18 cotranscribed genes. A two‐component regulatory system (Npun_F1277/Npun_F1278) directly upstream from the biosynthetic genes was identified through comparative genomics and is likely involved in scytonemin regulation. In this study, the response regulator (RR), Npun_F1278, was evaluated for its ability to regulate scytonemin biosynthesis using a mutant strain of N. punctiforme deficient in this gene, hereafter strain Δ1278. Following UVA radiation, the typical stimulus to initiate scytonemin biosynthesis, Δ1278 was incapable of producing scytonemin. A phenotypic characterization of Δ1278 suggests that aside from the ability to produce scytonemin, the deletion of the Npun_F1278 gene does not affect the cellular morphology, cellular differentiation capability, or lipid‐soluble pigment complement of Δ1278 compared to the wildtype. The mutant, however, had a slower specific growth rate under white light and produced ~2.5‐fold more phycocyanin per cell under UVA than the wildtype. Since Δ1278 does not produce scytonemin, this study demonstrates that the RR gene, Npun_F1278, is essential for scytonemin biosynthesis in N. punctiforme. While most of the evaluated effects of this gene appear to be specific for scytonemin, this regulator may also influence the overall health of the cell and phycobiliprotein synthesis, directly or indirectly. This is the first study to identify a regulatory gene involved in the biosynthesis of the sunscreen scytonemin and posits a link between cell growth, pigment synthesis, and sunscreen production.     

The extracellular-matrix-retaining cyanobacterium Nostoc verrucosum accumulates trehalose, but is sensitive to desiccation

The aquatic cyanobacterium Nostoc verrucosum forms macroscopic colonies, which consist of both cellular filaments and massive extracellular matrix material. In this study, the physiological features of N. verrucosum were investigated and compared with those of the anhydrobiotic cyanobacterium Nostoc commune. Nostoc verrucosum cells were sensitive to desiccation, but tolerant of freeze-thawing treatment in terms of both cell viability and photosynthetic O(2) evolution. Natural colonies of these cyanobacteria contained similar levels of chlorophyll a, carotenoids, the UV-absorbing pigments scytonemin and mycosporine-like amino acids, and uronic acid [a component of extracellular polysaccharides (EPS)]. EPS from both N. verrucosum and N. commune indicated low acidity and a high affinity for divalent cations, although their sugar compositions differed. The WspA protein, known to be a major component of the extracellular matrix of N. commune, was detected in N. verrucosum. Desiccation caused similarly high levels of trehalose accumulation in both cyanobacteria. Although previously considered relevant to anhydrobiosis in the terrestrial cyanobacterium N. commune, the data presented here suggest that extracellular matrix production and trehalose accumulation are not enough for standing extreme desiccation in N. verrucosum.     

Probing the in vivo biosynthesis of scytonemin, a cyanobacterial ultraviolet radiation sunscreen, through small scale stable isotope incubation studies and MALDI-TOF mass spectrometry

Scytonemin is a dimeric indole phenolic pigment found in the sheaths of many cyanobacteria. This pigment absorbs UV radiation protecting subtending cyanobacterial cells from harmful effects. Based on scytonemin's unique chemical structure, the pathway to its biosynthesis is uncertain, thus motivating the current investigation. Herein, we report the incorporation of both tyrosine and tryptophan into scytonemin, and provide in vivo data supporting the tryptophan origin of the ketone carbon involved in the condensation of the two biosynthetic precursors. This study also reports on the new use of a small-scale, MALDI-TOF mass spectrometry technique to monitor the incorporation of isotopically labeled tyrosine during scytonemin biosynthesis.     

Occurrence of UV-Absorbing, Mycosporine-Like Compounds among Cyanobacterial Isolates and an Estimate of Their Screening Capacity

A survey of 20 strains of cyanobacteria (belonging to 13 genera) isolated from habitats exposed to strong insolation revealed that 13 strains contained one or more water-soluble, UV-absorbing, mycosporine amino acid (MAA)-like compounds. Some of the compounds were identical in several strains. In all, 13 distinct compounds were found. The UV absorption spectra of MAAs complemented well that of the extracellular sunscreen pigment scytonemin, which many of the strains also produced. Even though the specific MAA contents were variable among strains, they were invariably higher when the cultures were grown with UV radiation than when it was absent. In five strains tested, the MAA complement accumulated as a solute in the cytoplasmic cell fraction. The sunscreen capacities of MAA and scytonemin and their combined capacity were estimated for each strain and condition on the basis of the specific contents, cell size, and cellular location of the compounds. The estimates suggested that significant, albeit not complete, protection from UV photodamage could be gained from the possession of either MAA or scytonemin but especially from simultaneous screening by both types of compounds.     

SCYTONEMIN, A CYANOBACTERIAL SHEATH PIGMENT, PROTECTS AGAINST UVC RADIATION: IMPLICATIONS FOR EARLY PHOTOSYNTHETIC LIFE

During the Precambrian, ultraviolet (UV) radiation reaching the Earth's surface, including UVC wavelengths (190–280 nm), was considerably higher than present because of the lack of absorbing gases (e.g. O₂ and O₃) in the atmosphere. High UV flux would have been damaging to photosynthetic organisms exposed to solar radiation. Nevertheless, fossil evidence indicates that cyanobacteria-like ancestors may have evolved as early as 3.5 × 10⁹ yr ago, and were common in shallow marine habitats by 2.5 × 10⁹ years ago. Scytonemin, a cyanobacterial extracellular sheath pigment, strongly absorbs UVC radiation. Exposure to high-irradiance conditions caused cells to synthesize scytonemin and resulted in decreased UVC inhibition of photosynthetic carbon uptake. It was further demonstrated that scytonemin alone was sufficient for substantial protection against UVC damage. This represents the first experimental demonstration of biological protection against UVC radiation in cyanobacteria. These results suggest that scytonemin may have evolved during the Precambrian and allowed colonization of exposed, shallow-water and terrestrial habitats by cyanobacteria or their oxygenic ancestors.     

Effect of environmental factors on the synthesis of scytonemin, a UV-screening pigment, in a cyanobacterium (Chroococcidiopsis sp.)

Abstract.The UV-screening pigment scytonemin is found in many species of ensheathed cyanobacteria. Past work has shown that the pigment is synthesized in response to exposure to UV-A irradiance. This study investigated the effect of other correlated stress factors including heat, osmotic and oxidative stress on the synthesis of scytonemin in a clonal cyanobacterial isolate ( Chroococcidiopsis sp.) from an epilithic desert crust. Stress experiments were carried out both in conjunction with UV-A irradiance and in isolation. Increases in both temperature and photooxidative conditions in conjunction with UV-A caused a synergistic increase in the rate of scytonemin production. In contrast, increased salt concentration under UV-A irradiance inhibited scytonemin synthesis. However, unlike the responses to temperature and oxidative stress, cells synthesized low levels of scytonemin under osmotic stress in the absence of scytonemin-inducing irradiance. These results suggest that scytonemin induction may be regulated as a part of a complex stress response pathway in which multiple environmental signals affect its synthesis.     

Effects of different UV radiation on photoprotective pigments and antioxidant activity of the hot‐spring cyanobacterium Leptolyngbya cf. fragilis

UV‐induced synthesis/accumulation of photoprotective pigments and antioxidant activity were investigated in the hot‐spring cyanobacterium Leptolyngbya cf. fragilis. The results indicated that UV radiation may induce biosynthesis of carotenoids, allophycocyanin, phycoerythrin, and scytonemin while phycocyanin degrades in response to longtime UV radiation. Moreover, pigment composition of L. cf. fragilis was significantly altered with increasing UV radiation times, probably due to destruction and resynthesis of accessory pigments as an adaptation strategy to UV stress. The in vitro antioxidant analysis of different extracts of UV treated cyanobacteria exhibited concentration‐dependent antioxidant activity. Ethyl acetate extract of 72 h UV treatment showed maximum total antioxidant activity (IC50 = 71.73 ± 5.3 μg mL^?1) followed by ethyl acetate control (non‐UV irradiated) extract (IC50 = 109.43 ± 2.76 μg mL^?1). This is the first report for the UV‐induced synthesis of photoprotective pigments and their antioxidant activity in L. cf. fragilis.     

Prodigiosin from <Emphasis Type="BoldItalic">Vibrio</Emphasis> sp. DSM 14379; A New UV-Protective Pigment

Pigments such as melanin, scytonemin and carotenoids protect microbial cells against the harmful effects of ultraviolet (UV) radiation. The role in UV protection has never been assigned to the prodigiosin pigment. In this work, we demonstrate that prodigiosin provides a significant level of protection against UV stress in Vibrio sp. DSM 14379. In the absence of pigment production, Vibrio sp. was significantly more susceptible to UV stress, and there was no difference in UV survival between the wild-type strain and non-pigmented mutant. The pigment’s protective role was more important at higher doses of UV irradiation and correlated with pigment concentration in the cell. Pigmented cells survived high UV exposure (324 J/m²) around 1,000-fold more successfully compared to the non-pigmented mutant cells. Resistance to UV stress was conferred to the non-pigmented mutant by addition of exogenous pigment extract to the growth medium. A level of UV protection equivalent to that exhibited by the wild-type strain was attained by the non-pigmented mutant once the prodigiosin concentration had reached comparable levels to those found in the wild-type strain. In co-culture experiments, prodigiosin acted as a UV screen, protecting both the wild-type and non-pigmented mutants. Our results suggest a new ecophysiological role for prodigiosin.     

Field relationships between scytonemin density,growth, and irradiance in cyanobacteria occurring in low illumination regimes

In situ measurements of ultraviolet (UV) irradiance, carbon fixation, and scytonemin pigmentation were made on Scytonema populations from contrasting localities in England. Significant negative correlations were obtained between the following variate pairs: pigmentation and UV irradiance; pigmentation and carbon fixation rate. A significant positive correlation was found between pigmentation and sheath thickness.The negative correlation between pigmentation and UV irradiance was unexpected and appeared contrary to the results of previous studies, which indicated a positive correlation between the variates and the recognition of scytonemin as a radiation shield. However, by considering how radiation damage is related to cell division and the water relations of the sites investigated, it was shown that scytonemin is still functioning as a radiation shield, even in shaded sites. Rivularia colonies produced scytonemin only upon their upper, sun-exposed surfaces and were positively correlated with UV irradiance. This paper also describes the successful use of some new and inexpensive techniques to measure pigments in cyanobacterium sheaths, and integrated in situ UV-irradiance.     

The biosynthesis of cyanobacterial sunscreen scytonemin in intertidal microbial mat communities

We have examined the biosynthesis and accumulation of cyanobacterial sunscreening pigment scytonemin within intertidal microbial mat communities using a combination of chemical, molecular, and phylogenetic approaches. Both laminated (layered) and nonlaminated mats contained scytonemin, with morphologically distinct mats having different cyanobacterial community compositions. Within laminated microbial mats, regions with and without scytonemin had different dominant oxygenic phototrophs, with scytonemin-producing areas consisting primarily of Lyngbya aestuarii and scytonemin-deficient areas dominated by a eukaryotic alga. The nonlaminated mat was populated by a diverse group of cyanobacteria and did not contain algae. The amplification and phylogenetic assignment of scytonemin biosynthetic gene scyC from laminated mat samples confirmed that the dominant cyanobacterium in these areas, L. aestuarii, is likely responsible for sunscreen production. This study is the first to utilize an understanding of the molecular basis of scytonemin assembly to explore its synthesis and function within natural microbial communities.     

A comprehensive study of eco-friendly natural pigment and its applications

Actinomycetes, a large group of filamentous bacteria account for 70–80% of secondary metabolites available commercially. The present investigation was undertaken with an aim to identify and characterize pigment from actinomycetes. Actinomycetes were isolated from rhizosphere soil samples collected from different regions of Madhya Pradesh state. Out of 85 actinomycetes, only 5 actinomycetes showed pigment production and based on diffusible pigment production ability one actinomycete ARITM02 was selected. The extraction of pigment was done by solvent extraction method using methanol and purified by TLC and column chromatography. The pigment was characterized by UV–Vis spectroscopy which showed the lamda maximum of 277.44. FTIR spectroscopy suggested various functional groups like amino group, amide group, hydroxide, benzene and lactone group. The Mass spectroscopy and NMR spectroscopy showed that the molecular mass of pigment is 621.7 and molecular formula is C34H43N3O8. The pigment was also tested for Antimicrobial activity against broad spectrum human pathogens, antioxidant test and toxicity test for safe use as a natural colorant in cosmetic, food, pharmaceutical and textile industries. The conclusion of study suggested that this novel pigment could be a versatile natural, safe and multipurpose.