Mycosporine-like Amino Acids Research Articles

Recent Advances and Future Prospects of Mycosporine-like Amino Acids.

Mycosporine-like amino acids (MAAs) are a class of water-soluble active substances produced by various aquatic organisms. However, due to the limitations of low accumulation of MAAs in organisms, the cumbersome extraction process, difficult identification, and high cost, MAAs have not yet been widely used in human life. Recently, there has been an emergence of heterologous synthesis for MAAs, making increasing yield the key to the quantification and application of MAAs. This review summarizes the latest research progress of MAAs, including: (1) introducing the biodistribution of MAAs and the content differences among different species to provide a reference for the selection of research subjects; (2) elaborating the species and molecular information of MAAs; (3) dissecting the synthesis mechanism and sorting out the synthesis pathways of various MAAs; (4) summarizing the methods of extraction and identification, summarizing the advantages and disadvantages, and providing a reference for the optimization of extraction protocols; (5) examining the heterologous synthesis method; and (6) summarizing the physiological functions of MAAs. This paper comprehensively updates the latest research status of MAAs and the various problems that need to be addressed, especially emphasizing the potential advantages of heterologous synthesis in the future production of MAAs.

Application of Synthetic Biology Approaches to High-Yield Production of Mycosporine-like Amino Acids

Ultraviolet (UV) radiation reaching the Earth’s surface is a major societal concern, and therefore, there is a significant consumer demand for cosmetics formulated to mitigate the harmful effects of UV radiation. Synthetic sunscreens being formulated to block UV penetration include inorganic metal oxide particles and organic filters. Lately, organic UV-absorbing compounds are manufactured from non-renewable petrochemicals and, as a result, there is a need to develop a sustainable manufacturing process for efficient, high-level production of a naturally occurring group of UV-absorbing compounds, namely mycosporine-like amino acids (MAAs), for use as a sunscreen additive to skincare products. Currently, the commercial production of MAAs for use in sunscreens is not a viable proposition due to the low yield and the lack of fermentation technology associated with native MAA-producing organisms. This review summarizes the biochemical properties of MAAs, the biosynthetic gene clusters and transcriptional regulations, the associated carbon-flux-driving processes, and the host selection and biosynthetic strategies, with the aim to expand our understanding on engineering suitable cyanobacteria for cost-effective production of natural sunscreens in future practices.

Gadusol is a maternally provided sunscreen that protects fish embryos from DNA damage

Exposure to ultraviolet radiation (UVR) is harmful to living cells, leading organisms to evolve protective mechanisms against UVR-induced cellular damage and stress.1,2 UVR, particularly UVB (280-320nm), can damageproteins and DNA, leading to errors during DNA repair and replication. Excessive UVR can induce cellulardeath. Aquatic organisms face risk of UV exposure as biologically harmful levels of UVB can penetrate >10m in clear water.3 While melanin is the only known sunscreen in vertebrates, it often emerges late in embryonic development, rendering embryos of many species vulnerable during the earlier stages. Algae and microbes produce a class of sunscreening compounds known as mycosporine-like amino acids (MAAs).4 Fish eggs contain a similar compound called gadusol, whose role as a sunscreen has yet to be tested despite its discovery over 40 years ago.5 The recent finding that many vertebrate genomescontain a biosynthetic pathway for gadusol suggests that many fish may produce and use this molecule as a sunscreen.6 We generated a gadusol-deficient mutant zebrafish to investigate the role of gadusol in protecting fish embryos and larvae from UVR. Our results demonstrate that maternally provided gadusol is the primary sunscreen in embryonic and larval development, while melanin provides modest secondary protection. The gadusol biosynthetic pathway is retained in the vast majority of teleost genomes but is repeatedly lost in species whose young are no longer exposed to UVR. Our data demonstrate that gadusol is a maternally provided sunscreen that is critical for early-life survival in the most species-rich branch of the vertebrate phylogeny.

Environmental adaptations by the intertidal Antarctic cyanobacterium Halotia branconiiCENA392 as revealed using long‐read genome sequencing

AbstractAntarctica poses numerous challenges to life such as cold shock, low nutrient concentrations, and periodic desiccation over a wide range of extreme temperatures. Cyanobacteria survive this harsh environment having evolved adaptive metabolic plasticity to become the dominant primary producers. The type strain cyanobacterium Halotia branconii CENA392 was isolated from an Antarctic intertidal seashore. The complete circular genome of this strain is presented herein, which was assembled using long‐sequence reads. The genome encoded some stress‐related genes associated with low‐temperature adaptation and biosynthesis of mycosporine‐like amino acid (MAA) photoprotective compounds. Empirical experimentation demonstrated constitutive production of the MAA porphyra‐334 and total carotenoids without exposure to low temperatures or ultraviolet radiation stress. Phylogenetic analysis provided insights on taxonomic placement and evolutionary history of some annotated genes. These data exemplify the importance of generating complete quality genome sequences of microorganisms isolated from extreme intertidal environments, facilitating in‐depth evaluation of ecological and taxonomic inferences.

Isolation of Mycosporine-like Amino Acids from Red Macroalgae and a Marine Lichen by High-Performance Countercurrent Chromatography: A Strategy to Obtain Biological UV-Filters

Marine organisms have gained considerable biotechnological interest in recent years due to their wide variety of bioactive compounds with potential applications. Mycosporine-like amino acids (MAAs) are UV-absorbing secondary metabolites with antioxidant and photoprotective capacity, mainly found in organisms living under stress conditions (e.g., cyanobacteria, red algae, or lichens). In this work, five MAAs were isolated from two red macroalgae (Pyropia columbina and Gelidium corneum) and one marine lichen (Lichina pygmaea) by high-performance countercurrent chromatography (HPCCC). The selected biphasic solvent system consisted of ethanol, acetonitrile, saturated ammonium sulphate solution, and water (1:1:0.5:1; v:v:v:v). The HPCCC process for P. columbina and G. corneum consisted of eight separation cycles (1 g and 200 mg of extract per cycle, respectively), whereas three cycles were performed for of L. pygmaea (1.2 g extract per cycle). The separation process resulted in fractions enriched with palythine (2.3 mg), asterina-330 (3.3 mg), shinorine (14.8 mg), porphyra-334 (203.5 mg) and mycosporine-serinol (46.6 mg), which were subsequently desalted by using precipitation with methanol and permeation on a Sephadex G-10 column. Target molecules were identified by HPLC, MS, and NMR.

Efficient production of natural sunscreens shinorine, porphyra-334, and mycosporine-2-glycine in Saccharomyces cerevisiae

Mycosporine-like amino acids (MAAs) are promising natural sunscreens mainly produced in marine organisms. Until now, metabolic engineering efforts to produce MAAs in heterologous hosts have mainly focused on shinorine production, and the low production levels are still not suitable for industrial applications. In this study, we successfully developed Saccharomyces cerevisiae strains that can efficiently produce various disubstituted MAAs, including shinorine, porphyra-334, and mycosporine-2-glycine (M2G), which are formed by conjugating serine, threonine, and glycine to mycosporine-glycine (MG), respectively. We first generated an MG-producing strain by multiple integration of the biosynthetic genes from cyanobacteria and applying metabolic engineering strategies to increase sedoheptulose-7-phosphate pool, a substrate for MG production. Next, five mysD genes from cyanobacteria, which encode D-Ala-D-Ala ligase homologues that conjugate an amino acid to MG, were introduced into the MG-producing strain to determine the substrate preference of each MysD enzyme. MysDs from Lyngbya sp., Nostoclinckia, and Euhalothece sp. showed high specificity toward serine, threonine, and glycine, resulting in efficient production of shinorine, porphyra-334, and M2G, respectively. This is the first report on the production of porphyra-334 and M2G in S. cerevisiae. Furthermore, we identified that the substrate specificity of MysD was determined by the omega loop region of 43–45 amino acids predicted based on its structural homology to a D-Ala-D-Ala ligase from Thermus thermophilus involved in peptidoglycan biosynthesis. The substrate specificities of two MysD enzymes were interchangeable by swapping the omega loop region. Using the engineered strain expressing mysD from Lyngbya sp. or N. linckia, up to 1.53 g/L shinorine or 1.21 g/L porphyra-334 was produced by fed-batch fermentation in a 5-L bioreactor, the highest titer reported so far. These results suggest that S. cerevisiae is a promising host for industrial production of different types of MAAs, providing a sustainable and eco-friendly alternative for the development of natural sunscreens.

Resolving Confusion Surrounding d-Ala-d-Ala Ligase Catalysis in Cyanobacterial Mycosporine-Like Amino Acid (MAA) Biosynthesis.

Mycosporine-like amino acids (MAAs) are natural UV-absorbing sunscreens that evolved in cyanobacteria and algae to palliate harmful effects from obligatory exposure to solar radiation. Multiple lines of evidence prove that in cyanobacteria all MAAs are derived from mycosporine-glycine, which is typically modified by an ATP-dependent ligase encoded by the gene mysD. The function of the mysD ligase has been experimentally described but haphazardly named based solely upon sequence similarity to the d-alanine-d-alanine ligase of bacterial peptidoglycan biosynthesis. Combining phylogeny and alpha-fold tertiary protein structure prediction unambiguously distinguished mysD from d-alanine-d-alanine ligase. The renaming of mysD to mycosporine-glycine-amine ligase (MG-amine ligase) using recognised enzymology rules of nomenclature is, therefore, proposed, and considers relaxed specificity for several different amino acid substrates. The evolutionary and ecological context of MG-amine ligase catalysis merits wider appreciation especially when considering exploiting cyanobacteria for biotechnology, for example, producing mixtures of MAAs with enhanced optical or antioxidant properties.

Exploring Mycosporine-like Amino Acid UV-Absorbing Natural Products for a New Generation of Environmentally Friendly Sunscreens.

Human skin needs additional protection from damaging ultraviolet radiation (UVR: 280-400 nm). Harmful UVR exposure leads to DNA damage and the development of skin cancer. Available sunscreens offer chemical protection from detrimental sun radiation to a certain extent. However, many synthetic sunscreens do not provide sufficient UVR protection due to the lack of photostability of their UV-absorbing active ingredients and/or the lack of ability to prevent the formation of free radicals, inevitably leading to skin damage. In addition, synthetic sunscreens may negatively affect human skin, causing irritation, accelerating skin aging and even resulting in allergic reactions. Beyond the potential negative effect on human health, some synthetic sunscreens have been shown to have a harmful impact on the environment. Consequently, identifying photostable, biodegradable, non-toxic, and renewable natural UV filters is imperative to address human health needs and provide a sustainable environmental solution. In nature, marine, freshwater, and terrestrial organisms are protected from harmful UVR through several important photoprotective mechanisms, including the synthesis of UV-absorbing compounds such as mycosporine-like amino acids (MAAs). Beyond MAAs, several other promising, natural UV-absorbing products could be considered for the future development of natural sunscreens. This review investigates the damaging impact of UVR on human health and the necessity of using sunscreens for UV protection, specifically UV-absorbing natural products that are more environmentally friendly than synthetic UV filters. Critical challenges and limitations related to using MAAs in sunscreen formulations are also evaluated. Furthermore, we explain how the genetic diversity of MAA biosynthetic pathways may be linked to their bioactivities and assess MAAs' potential for applications in human health.

Improvement of the UV-resistance capability of fish gelatin-oxidized starch film via inserting mycosporine-like amino acids.

Mycosporine-like amino acids (MAAs) are known as the strongest solar guardians in nature. In the present study, the extraction of MAAs from dried Pyropia haitanensis was achieved. Fish gelatin and oxidized starch composite films embedded with MAAs (0-0.3% w/w) were fabricated. The maximum absorption wavelength of the composite film appeared at 334 nm, which was consistent with MAA solution. Furthermore, the UV absorption intensity of the composite film was highly dependent on the concentration of MAAs. The composite film exhibited excellent stability during the 7-day storage period. The physicochemical features of composite film were demonstrated by the measurement of water content, water vapor transmission rate, oil transmission, and visual characteristics. Furthermore, in the actual anti-UV effect investigation, the increase in peroxide value and the acid value of grease under the films coverage was delayed. In the meantime, the decrease in ascorbic acid content in dates was postponed, and survivability of Escherichia coli was increased. Our results suggest that fish gelatin-oxidized starch-mycosporine-like amino acids film (FOM film) with biodegradable and anti-ultraviolet properties has a high potential for usage in food packaging materials. © 2023 Society of Chemical Industry.

Photosynthetic pigments and photoprotective metabolites of Colombian pacific marine macroalgae in response to contrasting ultraviolet-index periods

Tropical macroalgae have developed specific defense mechanisms, synthesizing photoprotective molecules to cope with solar radiations, especially ultraviolet radiation, which hits them intensely. Despite the vast Colombian marine biodiversity, not all the country's coastlines have received the same attention, like the Pacific coast. In this study, we assessed photosynthetic pigments (chlorophylls and carotenoids) and photoprotective compounds (mycosporine-like amino acids - MAAs) in seaweeds from the Colombian Pacific coast. We collected seven species of seaweeds, namely Boodleopsis verticillata, Bostrychia calliptera, B. radicans, B. cf. scorpioides; Catenella impudica, Cladophora graminea and Rhizoclonium riparium, easily found in supralittoral and mangrove ecosystems, at low tide, in contrasting periods of high and low UV index over the year of 2021. The sampling area encompassed a latitudinal gradient (3° S to 6° S) along the Colombian Pacific coast, changing the pattern of the chlorophyll-a and total carotenoid content in different species collected in contrasting UV-index periods. MAAs were quantified using standards (shinorine, porphyra-334, and palythine). This study corroborates the hypothesis that tropical species produce higher concentrations of MAAs in response to UV radiation, with significant differences in the high UV-index period. The highest MAA content (5.68 mg.g−1 DW) was reported in Bostrychia calliptera during the high UV-index period. Ceramiales and Cladophorales species represented a photoprotective potential with at least six different MAAs. Shinorine, palythine and porphyra-334 were quantified; and mycosporine-methylamine-serine and aplysiapalythine C were putatively identified by LC-MS/MS dereplication, this, therefore representing a pioneer study on the chemical composition of Colombian Pacific marine macroalgae.

Extraction of Mycosporine-like Amino Acids and Proteins from the Agarophyte Gelidium corneum Using Pulsed Power Techniques

Gelidium corneum (syn. sesquipedale) is an industrially and ecologically important species of red alga used for the production of high-quality agar. However, the species is also of growing interest for the production of other valuable compounds, such as mycosporine-like amino acids (MAAs), with potential cosmeceutical and biomedical applications. Novel methods using two pulsed power techniques, high-voltage electrical discharges (HVED) and pulsed electrical fields (PEF), were evaluated for efficacy of MAA extraction. Algal suspensions were prepared at two ratios (1:20 and 1:40 w:v). Four different extraction protocols were compared: (i) high-voltage electrical discharges, (ii) pulsed electric fields, (iii) maceration at room temperature, and (iv) maceration at 50 °C. The algae were treated in three states: freshly harvested, dried, and powdered. HVED and PEF treatments were effective when performed on fresh algae, and in particular the HVED treatment resulted in yields of MAAs twenty times higher than the control: 0.81 ± 0.05 mg/gDry Weight (DW) vs. 0.037 ± 0.002 mg/gDW. This effect was not observed to the same extent when the algae were dried or powdered, although HVED remained the most selective method overall.

Green Synthesis of Silver Nanoparticles and Its Combination with Pyropia columbina (Rhodophyta) Extracts for a Cosmeceutical Application.

We report the green synthesis of silver nanoparticles (AgNPs) by using daisy petals (Bellis perennis), leek (Allium porrum) and garlic skin (Allium sativum) as reducing agents and water as solvent. AgNPs are obtained with high monodispersity, spherical shapes and size ranging from 5 to 35 nm and characterized by UV-Vis and TEM techniques. The obtained yields in AgNPs are in concordance with the total phenolic content of each plant. We also study the incorporation of AgNPs in combination with the red algae Pyropia columbina extracts (PCE) into cosmetic formulations and analyze their combined effect as photoprotective agents. Moreover, we carry out the inclusion of the PCE containing mycosporine-like amino acids (MAAs), which are strong UV-absorbing and antioxidant compounds, into β-cyclodextrin (βCD) and pNIPAM nanoparticles and analyze stability and release. The thermoresponsive polymer is grown by free radical polymerization using N-isopropylacrylamide (NIPAM) as the monomer, N,N'-methylenebisacrylamide (BIS) as the cross-linker, and 2,2'-azobis(2-methylpropionamidene) (V50) as the initiator, while βCD complex is prepared by heating in water. We evaluate the nanoparticle and βCD complex formation by UV-Vis and FT-IR, and NMR spectroscopies, respectively, and the nanoparticles' morphology, including particle size, by TEM. The cosmetic formulations are subsequently subjected to accelerated stability tests and photoprotective analyses: a synergistic effect in the combination of AgNPs and PCE in photoprotection was found. It is not related to a UV screen effect but to the antioxidant activity, having potential against photoaging.

Marine Natural Products as Innovative Cosmetic Ingredients.

Over the course of the last 20 years, numerous studies have identified the benefits of an array of marine natural ingredients for cosmetic purposes, as they present unique characteristics not found in terrestrial organisms. Consequently, several marine-based ingredients and bioactive compounds are under development, used or considered for skin care and cosmetics. Despite the multitude of cosmetics based on marine sources, only a small proportion of their full potential has been exploited. Many cosmetic industries have turned their attention to the sea to obtain innovative marine-derived compounds for cosmetics, but further research is needed to determine and elucidate the benefits. This review gathers information on the main biological targets for cosmetic ingredients, different classes of marine natural products of interest for cosmetic applications, and the organisms from which such products can be sourced. Although organisms from different phyla present different and varied bioactivities, the algae phylum seems to be the most promising for cosmetic applications, presenting compounds of many classes. In fact, some of these compounds present higher bioactivities than their commercialized counterparts, demonstrating the potential presented by marine-derived compounds for cosmetic applications (i.e., Mycosporine-like amino acids and terpenoids' antioxidant activity). This review also summarizes the major challenges and opportunities faced by marine-derived cosmetic ingredients to successfully reach the market. As a future perspective, we consider that fruitful cooperation among academics and cosmetic industries could lead to a more sustainable market through responsible sourcing of ingredients, implementing ecological manufacturing processes, and experimenting with inventive recycling and reuse programs.

Characterization, UV-induction, antioxidant function and role in photo-protection of mycosporine-like amino acids (MAAs) in a unicellular cyanobacterium, Euhalothece sp.WR7

Mycosporine-like amino acids (MAAs) are considered an important and unique UV-absorbing biomolecules. The present study was intended to explore the occurrence, UV-induction, free radical scavenging potential and photoprotective function of MAAs in the cyanobacterium, Euhalothece sp.WR7. High-performance liquid chromatography (HPLC) coupled with photodiode-array (PDA) detection studies revealed the occurrence of two different UV-absorbing compounds having absorption maxima at 331 nm and 362 nm with retention time 6.4 and 10.2 min, respectively. Based on the HPLC-PDA detection and ion trap liquid chromatography-mass spectrometry (LC-MS) analysis, the compounds were identified as mycosporine-2-Glycine (M2G: λmax 331 nm, m/z 303) and Euhalothece-362 (E-362: λmax 362 nm, m/z 331). Compared to the photosynthetically active radiation, a substantial induction of the synthesis of both MAAs was observed under UV (UV-A + UV-B) stress condition. MAAs (M2G + E-362) showed significant free radical scavenging potential in a dose-dependent manner. It also exhibited efficient photoprotective function by diminishing the UV-induced generation of reactive oxygen species (ROS) and enhancing the survival of Escherichia coli cells under UV-B stress. The results indicate the dynamic role of MAAs in existence and viability of cyanobacterium by regulating the key ecological functions in a harsh environment with intense solar UV irradiances. Moreover, MAAs may be used as important UV absorbers in cosmetic sunscreens due to their strong photoprotective function.

Rapid Screening for Mycosporine-like Amino Acids (MAAs) of Irish Marine Cyanobacteria and Their Antioxidant Potential

The present study evaluated 53 Irish marine cyanobacteria intending to identify potential producers of mycosporine-like amino acids (MAAs) to meet the increasing demand for replacing chemical sunscreen with bio-sunscreen. The biodiscovery analysis using absorption spectra of methanolic extracts identified eight cyanobacteria as potential MAAs producers with a specific content of 0.114–0.511 A* mg DW−1. Leptolyngbya tenuis SABC010201 was found to possess notably higher MAAs content. LC-MS analysis identified a total of eight different types of known MAAs (mycosporine-glutamicol, mycosporine-glutaminol-glucoside, mycosporine-serinol, mycosporine-taurine, palythine, palythine-threonine-sulphate, porphyra-334, and usujirene) in eight cyanobacteria, while four compounds were considered unknown UV-absorbing compounds with specific mass and absorption maximum. For example, two unknown compounds with, respectively, [M-H]- values of 219.0557 and 289.0730 and lambda max of 314 and 326 nm, were detected in three cyanobacteria Leptolyngbya tenuis SABC010201, Phormidium angustissimum SABC020801, and Schizothrix sp. SABC022401. These two unknown compounds were named M-314 and M-326, respectively. Antioxidant activities of total MAAs of all cyanobacteria showed considerable amounts of DPPH, FRAP, and ORAC activities. Considering the specific MAAs content and antioxidant activities, Leptolyngbya africana SABC021601 was considered the best producer of MAAs.

Monthly Variation in Mycosporine-like Amino Acids from Red Alga Dulse (Devaleraea inkyuleei, Formerly Palmaria palmata in Japan)

Mycosporine-like amino acids (MAAs) are natural ultraviolet-absorbing compounds found in microalgae and macroalgae. MAA content changes seasonally and in response to environmental factors. We previously investigated MAAs from the red alga dulse (Devaleraea inkyuleei, formerly Palmaria palmata in Japan) in Usujiri, Hokkaido, Japan, from 2019 to 2020. At that time, some factors affecting MAA content were still unclear. In this study, we investigated MAA variation during the period from January to June 2021, and evaluated new methods of MAA extraction from dulse. We recorded a maximum MAA extraction yield (7.03 µmol/g dry weight) on 25 March 2021. Over the course of our three years of investigations from 2019 to 2021, we found that dulse was most suitable for MAA preparation from the middle of February to late April. In the later work reported in this paper, we improved our extraction method by using a lower-risk organic solvent (ethanol) rather than methanol. In addition, we evaluated MAA extraction using different levels of ethanol concentration (25, 50, and 99%) and different extraction times (2, 6, and 24 h). We found that extraction with 25% ethanol for 24 h increased MAA content by a factor of 3.2, compared with our previous extraction method. In summary, we determined the most suitable sampling period for Usujiri dulse, to extract the highest content of MAAs. We also improved the effectiveness of the extraction process.

Separating and Purifying Mycosporine-like Amino Acids from Cyanobacteria for Application in Commercial Sunscreen Formulations.

Using algal-derived mycosporine-like amino acids (MAAs) in sunscreen formulations is constrained by low cellular concentrations of MAAs and by the high costs associated with harvesting algal cells and extracting the MAAs. Here, we report an industrial scalable method using a membrane filtration approach to purify and concentrate aqueous extracts of MAAs. The method includes an additional biorefinery step enabling purification of phycocyanin, an established valuable natural product. Cultivated cells of the cyanobacterium Chlorogloeopsis fritschii (PCC 6912) were concentrated and homogenised to produce a feed for sequential processing through three membranes of decreasing pore size to obtain a retentate and permeate for each step. Microfiltration (0.2 µm) was used to remove cell debris. Ultrafiltration (10,000 Da) was used to remove large molecules and recover phycocyanin. Finally, nanofiltration (300-400 Da) was used to remove water and other small molecules. Permeate and retentate were analysed using UV-visible spectrophotometry and HPLC. The initial homogenised feed had a shinorine concentration of 5.6 ± 07 mg L-1. The final nanofiltered retentate resulted in a 3.3 times-purified concentrate (shinorine concentration of 18.71 ± 0.29 mg L-1). Significant process losses (35%) highlight scope for improvement. Results confirm the potential of membrane filtration to purify and concentrate aqueous solutions of MAAs with simultaneous separation of phycocyanin highlighting a biorefinery approach.

Exploring the Relationship between Biosynthetic Gene Clusters and Constitutive Production of Mycosporine-like Amino Acids in Brazilian Cyanobacteria.

Cyanobacteria are oxygenic phototrophic prokaryotes that have evolved to produce ultraviolet-screening mycosporine-like amino acids (MAAs) to lessen harmful effects from obligatory exposure to solar UV radiation. The cyanobacterial MAA biosynthetic cluster is formed by a gene encoding 2-epi-5-epi-valiolone synthase (EVS) located immediately upstream from an O-methyltransferase (OMT) encoding gene, which together biosynthesize the expected MAA precursor 4-deoxygadusol. Accordingly, these genes are typically absent in non-producers. In this study, the relationship between gene cluster architecture and constitutive production of MAAs was evaluated in cyanobacteria isolated from various Brazilian biomes. Constitutive production of MAAs was only detected in strains where genes formed a co-linear cluster. Expectedly, this production was enhanced upon exposure of the strains to UV irradiance and by using distinct culture media. Constitutive production of MAAs was not detected in all other strains and, unexpectedly, production could not be induced by exposure to UV irradiation or changing growth media. Other photoprotection strategies which might be employed by these MAA non-producing strains are discussed. The evolutionary and ecological significance of gene order conservation warrants closer experimentation, which may provide a first insight into regulatory interactions of genes encoding enzymes for MAA biosynthesis.

Halotolerance, stress mechanisms, and circadian clock of salt-tolerant cyanobacteria.

Cyanobacteria harbor a high level of physiological flexibility, which enables them to reside in virtually all available environmental niches, including extreme environments. In this review, we summarize the recent advancements in stress mechanisms of salt-tolerant (a.k.a. halotolerant) cyanobacteria. Omics approaches have been extensively employed in recent years to decipher mechanisms of halotolerance and to understand the relevance of halotolerance-associated gene regulatory networks. The vast knowledge from genome mining disclosed that halotolerant cyanobacteria possess extended gene families and/or clusters, encoding enzymes that synthesize unique osmoprotectants, including glycine betaine (GB), betaine derivatives, and mycosporine-like amino acids (MAAs). Comprehensive transcriptomic analyses were conducted using Halothece sp. PCC7418 (hereafter referred to as Halothece), a cyanobacterium that exhibits remarkable halotolerance. These studies revealed a specific transcriptional response when Halothece was subjected to salt stress, whereas salt and osmotic stresses were found to share a common transcriptomic response. Transcriptome and metabolite analyses of Halothece illustrated a complex dynamic relationship between the biosyntheses of osmoprotectants, as well as corresponding and ancillary pathways. Lastly, novel insights highlight the relationship between the molecular regulation of the circadian rhythm and salt stress tolerance. Since the circadian rhythm of gene expression was distorted under salt stress, halotolerant cyanobacteria may prioritize the adaptation to salt stress by attenuation of circadian rhythmicity. KEY POINTS: • Recent advancements in the understanding of stress mechanisms in halotolerant cyanobacteria are described based on omics analyses. • Transcriptome and metabolite analyses of Halothece illustrated a complex dynamic relationship between the biosyntheses of osmoprotectants, as well as corresponding and ancillary pathways. • Since salt stress affects the molecular regulation among clock-related proteins, salt stress may attenuate circadian rhythmicity.

Application of feature-based molecular networking in the field of algal research with special focus on mycosporine-like amino acids

Marine red algae have been known as an excellent source for natural sunscreens and antioxidants for a long time, which outlines their potential for various medical and cosmeceutical applications. This is due to their synthesis of unique secondary metabolites to shield themselves from high levels of UV-A and -B radiation encountered in their natural habitats. In this study, a comprehensive and contemporary way for the detection, visualization, and dereplication of algal natural products with special focus on mycosporine-like amino acids (MAAs) is shown, employing HR-MS/MS metabolomics. 33 crude algal extracts were explored using ultra-high-performance liquid chromatography (UHPLC) hyphenated to orbitrap high-resolution tandem mass spectroscopy (HRMS2). Acquired raw data, subjected to pretreatment and spectral organization, could subsequently be implemented in the Global Natural Products Social (GNPS) workflow, whereby a feature based molecular network (FBMN) was created and visualized in Cytoscape. This FBMN was matched against an in-house as well as open source library on the GNPS platform and additionally enhanced by chemotaxonomic classification software and spectra of standard MAAs, as well as further information layers covering e.g. physicochemical properties, taxonomy, and fragmentation behavior. Based on the integration of the latest in silico annotation tools (SIRIUS, CANOPUS, MSNovelist) as well as already published fragmentation patterns of MAAs, structures for known compounds could be corroborated as well as those for novel substances proposed. This offers an interesting and state-of-the-art approach towards the identification and classification of known and new MAAs.