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1.
Plant Physiol Biochem ; 214: 108942, 2024 Jul 16.
Article in English | MEDLINE | ID: mdl-39024780

ABSTRACT

Eukaryotic microalgae are a diverse group of organisms that can be used for the sustainable production of a wide range of high value compounds, including lipids, flavours and dyes, bioplastics, and cosmetics. Optimising total biomass production often does not lead to optimal product yield and more sophisticated biphasic growth strategies are needed, introducing specific stresses to induce product synthesis. Genetic tools have been used to increase yields of natural products or to introduce new pathways to algae, and wider deployment of these tools offers promising routes for commercial production of high value compounds utilising minimal inputs.

2.
Mar Drugs ; 22(5)2024 May 15.
Article in English | MEDLINE | ID: mdl-38786613

ABSTRACT

Porphyra sensu lato has economic importance for food and pharmaceutical industries due to its significant physiological activities resulting from its bioactive compounds (BACs). This study aimed to determine the optimal nitrate dosage required in short-term cultivation to achieve substantial BAC production. A nitrate experiment using varied concentrations (0 to 6.5 mM) revealed optimal nitrate uptake at 0.5 mM in the first two days and at 3 and 5 mM in the last five days. Polyphenols and carbohydrates showed no differences between treatments, while soluble proteins peaked at 1.5 and 3 mM. Total mycosporine-like amino acids (MAAs) were highest in algae incubated at 5 and 6.5 mM, and the highest antioxidant activity was observed in the 5 mM, potentially related to the MAAs amount. Total carbon and sulfur did not differ between treatments, while nitrogen decreased at higher nitrate. This discovery highlights the nuanced role of nitrate in algal physiology, suggesting that biological and chemical responses to nitrate supplementation can optimize an organism's health and its commercially significant bioactive potential. Furthermore, given its ability to absorb high doses of nitrate, this alga can be cultivated in eutrophic zones or even in out-/indoor tanks, becoming an excellent option for integrated multi-trophic aquaculture (IMTA) and bioremediation.


Subject(s)
Antioxidants , Biodegradation, Environmental , Nitrates , Porphyra , Nitrates/metabolism , Nitrates/pharmacology , Antioxidants/pharmacology , Antioxidants/metabolism , Porphyra/metabolism , Cosmeceuticals , Amino Acids/metabolism
3.
Microb Cell Fact ; 23(1): 121, 2024 May 09.
Article in English | MEDLINE | ID: mdl-38725068

ABSTRACT

BACKGROUND: Mycosporine-like amino acids (MAAs) are a class of strongly UV-absorbing compounds produced by cyanobacteria, algae and corals and are promising candidates for natural sunscreen components. Low MAA yields from natural sources, coupled with difficulties in culturing its native producers, have catalyzed synthetic biology-guided approaches to produce MAAs in tractable microbial hosts like Escherichia coli, Saccharomyces cerevisiae and Corynebacterium glutamicum. However, the MAA titres obtained in these hosts are still low, necessitating a thorough understanding of cellular factors regulating MAA production. RESULTS: To delineate factors that regulate MAA production, we constructed a shinorine (mycosporine-glycine-serine) producing yeast strain by expressing the four MAA biosynthetic enzymes from Nostoc punctiforme in Saccharomyces cerevisiae. We show that shinorine is produced from the pentose phosphate pathway intermediate sedoheptulose 7-phosphate (S7P), and not from the shikimate pathway intermediate 3-dehydroquinate (3DHQ) as previously suggested. Deletions of transaldolase (TAL1) and phosphofructokinase (PFK1/PFK2) genes boosted S7P/shinorine production via independent mechanisms. Unexpectedly, the enhanced S7P/shinorine production in the PFK mutants was not entirely due to increased flux towards the pentose phosphate pathway. We provide multiple lines of evidence in support of a reversed pathway between glycolysis and the non-oxidative pentose phosphate pathway (NOPPP) that boosts S7P/shinorine production in the phosphofructokinase mutant cells. CONCLUSION: Reversing the direction of flux between glycolysis and the NOPPP offers a novel metabolic engineering strategy in Saccharomyces cerevisiae.


Subject(s)
Amino Acids , Glycolysis , Pentose Phosphate Pathway , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae/genetics , Amino Acids/metabolism , Metabolic Engineering/methods , Nostoc/metabolism , Nostoc/genetics , Sugar Phosphates/metabolism , Glycine/metabolism , Glycine/analogs & derivatives , Cyclohexylamines
4.
Biosci Biotechnol Biochem ; 88(7): 830-838, 2024 Jun 21.
Article in English | MEDLINE | ID: mdl-38684478

ABSTRACT

Mycosporine-like amino acids (MAAs) are the natural UV-absorbing compounds with antioxidant activity found in microalgae and macroalgae. We collected red algae Asparagopsis taxiformis, Meristotheca japonica, and Polysiphonia senticulosa from Nagasaki, where UV radiation is more intense than in Hokkaido, and investigated the effect of UV radiation on MAA content. It was suggested that A. taxiformis and M. japonica contained shinorine and palythine, while UV-absorbing compound in P. senticulosa could not be identified. The amounts of these MAAs were lower compared to those from Hokkaido. Despite an increase in UV radiation in both regions from February to April, MAA contents of red algae from Nagasaki slightly decreased while those from Hokkaido significantly decreased. This difference was suggested the amount of inorganic nitrogen in the ocean. Antioxidant activity of MAAs increased under alkaline conditions. The extract containing MAAs from P. senticulosa showed the highest antioxidant activity among 4 red algae.


Subject(s)
Amino Acids , Antioxidants , Rhodophyta , Rhodophyta/chemistry , Amino Acids/analysis , Antioxidants/chemistry , Antioxidants/pharmacology , Japan , Ultraviolet Rays , Biphenyl Compounds/antagonists & inhibitors , Hydrogen-Ion Concentration , Cyclohexanols , Cyclohexylamines , Glycine/analogs & derivatives
5.
Mol Biotechnol ; 2024 Apr 23.
Article in English | MEDLINE | ID: mdl-38652428

ABSTRACT

Melanin is the major pigment responsible for the coloring of mammalian skin, hair, and eyes to defend against ultraviolet radiation. However, excessive melanin production has resulted in numerous types of hyperpigmentation disorders. Tyrosinase-related protein 1 (TYRP1) is a transmembrane glycoprotein enzyme found in many organisms, including humans, that plays an important role in melanogenesis. Thus, controlling the enzyme activity of TYRP1 with tyrosinase inhibitors is a vital step in the treatment of hyperpigmentation problems in humans. In the present investigation, virtual screening, pharmacokinetics, drug docking, and molecular dynamics (MD) simulation were used to find the most potent drug as an inhibitor of TYRP1 to effectively treat hyperpigmentation disorder. The 3D structure of TYRP1 was retrieved from the Protein Data Bank (PDB) database (PDB ID: 5M8M) and validated by the Ramachandran plot. Pharmacokinetics and drug-likeness showed that mycosporine 2 glycine (M2G) and shinorine (SHI) were the best compounds over other ligands in the same (P-1) structural pose. However, MD simulations of the M2G showed the highest CDOCKER interaction energy (-45.182 kcal/mol) and binding affinity (-65.0529 kcal/mol) as compared to SHI and reference drugs. The molecular binding modes RMSD and RMSF plots have exhibited more relevance to the M2G ligand in comparison to other drug ligands. The bioactivity and ligand efficiency profiles revealed that M2G is the most effective compound as a TYRP1 inhibitor. Thus, M2G could be used as a most effective drug for developing valuable sunscreen products to cure hyperpigmentation-related diseases.

6.
Photochem Photobiol ; 100(2): 298-313, 2024.
Article in English | MEDLINE | ID: mdl-37312642

ABSTRACT

Sunscreens provide a frontline defense for our DNA against the damage caused by ultraviolet (UV) radiation. The active ingredients in topically applied sunscreens that provide this defense are UV filters, which preferentially absorb or reflect UV radiation before it penetrates the skin and interacts with photosensitive nucleic acids. However, there are concerns related to human and environmental toxicity of current UV filters, and consequently a shift toward nature-inspired, particularly microbial, UV filters. In this paper, new physical insight is provided into the fundamental mechanisms of photoprotection in two synthetic analogs of mycosporine-like amino acid-type UV filters, demonstrating new methods of protection that are distinct from those of current commercial sunscreens, extending previous work in this area. Transient absorption measurements (both transient electronic absorption spectroscopy and transient vibrational absorption spectroscopy) are combined with steady-state studies and high-level computational results to aid our mapping of the experimentally derived lifetimes to real-time photodynamic processes. The conclusions reached here pave the way toward developing new and more efficient biomimetic DNA photoprotectant materials.


Subject(s)
Sunscreening Agents , Ultraviolet Rays , Humans , Sunscreening Agents/chemistry , Isomerism , Skin , DNA
7.
Mar Drugs ; 21(12)2023 Dec 08.
Article in English | MEDLINE | ID: mdl-38132954

ABSTRACT

Marine macroalgae (seaweeds) are important primary global producers, with a wide distribution in oceans around the world from polar to tropical regions. Most of these species are exposed to variable environmental conditions, such as abiotic (e.g., light irradiance, temperature variations, nutrient availability, salinity levels) and biotic factors (e.g., grazing and pathogen exposure). As a result, macroalgae developed numerous important strategies to increase their adaptability, including synthesizing secondary metabolites, which have promising biotechnological applications, such as UV-absorbing Mycosporine-Like Amino Acid (MAAs). MAAs are small, water-soluble, UV-absorbing compounds that are commonly found in many marine organisms and are characterized by promising antioxidative, anti-inflammatory and photoprotective properties. However, the widespread use of MAAs by humans is often restricted by their limited bioavailability, limited success in heterologous expression systems, and low quantities recovered from the natural environment. In contrast, bloom-forming macroalgal species from all three major macroalgal clades (Chlorophyta, Phaeophyceae, and Rhodophyta) occasionally form algal blooms, resulting in a rapid increase in algal abundance and high biomass production. This review focuses on the bloom-forming species capable of producing pharmacologically important compounds, including MAAs, and the application of proteomics in facilitating macroalgal use in overcoming current environmental and biotechnological challenges.


Subject(s)
Rhodophyta , Seaweed , Humans , Seaweed/chemistry , Ultraviolet Rays , Seasons , Amino Acids/chemistry , Rhodophyta/chemistry
8.
J Ind Microbiol Biotechnol ; 50(1)2023 Feb 17.
Article in English | MEDLINE | ID: mdl-37950572

ABSTRACT

Exposure to ultraviolet (UV) rays is a known risk factor for skin cancer, which can be notably mitigated through the application of sun care products. However, escalating concerns regarding the adverse health and environmental impacts of synthetic anti-UV chemicals underscore a pressing need for the development of biodegradable and eco-friendly sunscreen ingredients. Mycosporine-like amino acids (MAAs) represent a family of water-soluble anti-UV natural products synthesized by various organisms. These compounds can provide a two-pronged strategy for sun protection as they not only exhibit a superior UV absorption profile but also possess the potential to alleviate UV-induced oxidative stresses. Nevertheless, the widespread incorporation of MAAs in sun protection products is hindered by supply constraints. Delving into the biosynthetic pathways of MAAs can offer innovative strategies to overcome this limitation. Here, we review recent progress in MAA biosynthesis, with an emphasis on key biosynthetic enzymes, including the dehydroquinate synthase homolog MysA, the adenosine triphosphate (ATP)-grasp ligases MysC and MysD, and the nonribosomal peptide synthetase (NRPS)-like enzyme MysE. Additionally, we discuss recently discovered MAA tailoring enzymes. The enhanced understanding of the MAA biosynthesis paves the way for not only facilitating the supply of MAA analogs but also for exploring the evolution of this unique family of natural sunscreens. ONE-SENTENCE SUMMARY: This review discusses the role of mycosporine-like amino acids (MAAs) as potent natural sunscreens and delves into recent progress in their biosynthesis.


Subject(s)
Amino Acids , Sunscreening Agents , Amino Acids/chemistry , Sunscreening Agents/chemistry , Sunscreening Agents/pharmacology , Oxidative Stress , Ultraviolet Rays
9.
Mar Drugs ; 21(10)2023 Oct 18.
Article in English | MEDLINE | ID: mdl-37888478

ABSTRACT

This study presents a phytochemical survey of two common intertidal red algal species, Bostrychia scorpioides and Catenella caespitosa, regarding their MAA (mycosporine-like amino acid) composition, which are known as biogenic sunscreen compounds. Six novel MAAs from Bostrychia scorpioides named bostrychines and two novel MAAs from Catenella caespitosa named catenellines were isolated using a protocol which included silica gel column chromatography, flash chromatography on reversed phase material and semipreparative HPLC (High-Performance Liquid Chromatography). The structure of the novel MAAs was elucidated using NMR (Nuclear Magnetic Resonance) and HR-MS (High-Resolution Mass Spectrometry), and their absolute configuration was confirmed by ECD (Electronic Circular Dichroism). All isolated MAAs possess a cyclohexenimine scaffold, and the metabolites from B. scorpioides are related to the known MAAs bostrychines A-F, which contain glutamine, glutamic acid and/or threonine in their side chains. The new MAAs from C. caespitosa contain taurine, an amino sulfonic acid that is also present in another MAA isolated from this species, namely, catenelline. Previous and new data confirm that intertidal red algae are chemically rich in MAAs, which explains their high tolerance against biologically harmful ultraviolet radiation.


Subject(s)
Rhodophyta , Seaweed , Amino Acids/chemistry , Seaweed/chemistry , Ultraviolet Rays , Rhodophyta/chemistry , Chromatography, High Pressure Liquid
10.
Biotechnol Biofuels Bioprod ; 16(1): 162, 2023 Oct 29.
Article in English | MEDLINE | ID: mdl-37899467

ABSTRACT

BACKGROUND: Mycosporine-like amino acids (MAAs), including shinorine and porphyra-334, are gaining attention as safe natural sunscreens. The production of MAAs has been achieved in diverse microbial hosts, including Saccharomyces cerevisiae. While S. cerevisiae is the most extensively studied model yeast, the oleaginous yeast Yarrowia lipolytica has emerged as a promising candidate for the synthesis of valuable products. In this study, we explored the potential of Y. lipolytica as a host for producing MAAs, utilizing its advantages such as a robust pentose phosphate pathway flux and versatile carbon source utilization. RESULTS: We produced MAAs in Y. lipolytica by introducing the MAA biosynthetic genes from cyanobacteria Nostoc punctiforme and Anabaena variabilis. These genes include mysA, mysB, and mysC responsible for producing mycosporine-glycine (MG) from sedoheptulose 7-phosphate (S7P). The two strains utilize different enzymes, D-Ala-D-Ala ligase homologue (MysD) in N. punctiforme and NRPS-like enzyme (MysE) in A. variabilis, for amino acid conjugation to MG. MysE specifically generated shinorine, a serine conjugate of MG, while MysD exhibited substrate promiscuity, yielding both shinorine and a small amount of porphyra-334, a threonine conjugate of MG. We enhanced MAAs production by selecting mysA, mysB, and mysC from A. variabilis and mysD from N. punctiforme based on their activities. We further improved production by strengthening promoters, increasing gene copies, and introducing the xylose utilization pathway. Co-utilization of xylose with glucose or glycerol increased MAAs production by boosting the S7P pool through the pentose phosphate pathway. Overexpressing GND1 and ZWF1, key genes in the pentose phosphate pathway, further enhanced MAAs production. The highest achieved MAAs level was 249.0 mg/L (207.4 mg/L shinorine and 41.6 mg/L of porphyra-334) in YP medium containing 10 g/L glucose and 10 g/L xylose. CONCLUSIONS: Y. lipolytica was successfully engineered to produce MAAs, primarily shinorine. This achievement involved the introduction of MAA biosynthetic genes from cyanobacteria, establishing xylose utilizing pathway, and overexpressing the pentose phosphate pathway genes. These results highlight the potential of Y. lipolytica as a promising yeast chassis strain for MAAs production, notably attributed to its proficient expression of MysE enzyme, which remains non-functional in S. cerevisiae, and versatile utilization of carbon sources like glycerol.

11.
Appl Environ Microbiol ; 89(11): e0074023, 2023 11 29.
Article in English | MEDLINE | ID: mdl-37843273

ABSTRACT

Recent years have seen a lot of interest in mycosporine-like amino acids (MAAs) because of their alleged potential as a natural microbial sunscreen. Since chemical ultraviolet (UV) absorbers are unsafe for long-term usage, the demand for natural UV-absorbing substances has increased. In this situation, MAA is a strong contender for an eco-friendly UV protector. The capacity of MAAs to absorb light in the UV-A (320-400 nm) and UV-B (280-320 nm) range without generating free radicals is potentially relevant in photoprotection. The usage of MAAs for purposes other than photoprotection has now shifted in favor of medicinal applications. Aside from UV absorption, MAAs also have anti-oxidant, anti-inflammatory, wound-healing, anti-photoaging, cell proliferation stimulators, anti-cancer agents, and anti-adipogenic properties. Recently, MAAs application to combat SARS-CoV-2 infection was also investigated. In this review article, we highlight the biomedical applications of MAAs that go beyond photoprotection, which can help in utilizing the MAAs as promising bioactive compounds in both pharmaceutical and cosmetic applications.


Subject(s)
Amino Acids , Ultraviolet Rays , Amino Acids/metabolism , Anti-Inflammatory Agents , Sunscreening Agents/pharmacology , Sunscreening Agents/chemistry , Sunscreening Agents/metabolism , Antioxidants
12.
Int J Mol Sci ; 24(20)2023 Oct 10.
Article in English | MEDLINE | ID: mdl-37894736

ABSTRACT

Although it is well recognized that mycosporine-like amino acids (MAAs) are ultraviolet (UV) protective agents that can reduce UV damage, the specific biological mechanism of its role in the skin remains unclear. In this study, we investigated the effect of MAAs extracted from Antarctic diatom Phaeodactylum tricornutum ICE-H on UVB-induced skin damage using a mice model. The MAAs components identified by liquid chromatography-tandem mass spectrometry included 4-deoxygadusol, shinorine, and porphyra-334, which were purified using a Supledean Carboxen1000 solid phase extraction column. The antioxidant activities of these MAA compounds were tested in vitro. For UVB-induced skin photodamage in mice, MAAs alleviated skin swelling and epidermal thickening in this study. We detected the content of reactive oxygen species (ROS), malondialdehyde, and collagen in skin tissue. In addition, quantitative real-time polymerase chain reaction was used to detect nuclear factor-κB (NF-κB), tumor necrosis factor α, interleukin-1ß, cyclooxygenase-2, mitogen activated protein kinase (MAPK) family (extracellular signal-regulated kinase, c-Jun amino-terminal kinase, and p38 kinase), and matrix metalloproteinases. The expression of these cytokines and enzymes is related to inflammatory responses and collagen degradation. In comparison to the model group without MAA treatment, the MAA component decreased the concentration of ROS, the degree of oxidative stress in the skin tissue, and the expression of genes involved in the NF-κB and MAPK pathways. In summary, these MAA components extracted from Phaeodactylum tricornutum ICE-H protected against UVB-induced skin damage by inhibiting ROS generation, relieving skin inflammation, and slowing down collagen degradation, suggesting that these MAA components are effective cosmetic candidate molecules for the protection and therapy of UVB damage.


Subject(s)
Amino Acids , Diatoms , Animals , Mice , Amino Acids/chemistry , Diatoms/metabolism , Reactive Oxygen Species/metabolism , NF-kappa B/metabolism , Antarctic Regions , Skin/metabolism , Mitogen-Activated Protein Kinases/metabolism , Collagen/pharmacology , Ultraviolet Rays/adverse effects
13.
Int J Mol Sci ; 24(15)2023 Aug 03.
Article in English | MEDLINE | ID: mdl-37569755

ABSTRACT

Ultraviolet radiation (UVR) tends to damage key cellular machinery. Cells may adapt by developing several defence mechanisms as a response to such damage; otherwise, their destiny is cell death. Since cyanobacteria are primary biotic components and also important biomass producers, any drastic effects caused by UVR may imbalance the entire ecosystem. Cyanobacteria are exposed to UVR in their natural habitats. This exposure can cause oxidative stress which affects cellular morphology and vital processes such as cell growth and differentiation, pigmentation, photosynthesis, nitrogen metabolism, and enzyme activity, as well as alterations in the native structure of biomolecules such as proteins and DNA. The high resilience and several mitigation strategies adopted by a cyanobacterial community in the face of UV stress are attributed to the activation of several photo/dark repair mechanisms, avoidance, scavenging, screening, antioxidant systems, and the biosynthesis of UV photoprotectants, such as mycosporine-like amino acids (MAAs), scytonemin (Scy), carotenoids, and polyamines. This knowledge can be used to develop new strategies for protecting other organisms from the harmful effects of UVR. The review critically reports the latest updates on various resilience and defence mechanisms employed by cyanobacteria to withstand UV-stressed environments. In addition, recent developments in the field of the molecular biology of UV-absorbing compounds such as mycosporine-like amino acids and scytonemin and the possible role of programmed cell death, signal perception, and transduction under UVR stress are discussed.


Subject(s)
Cyanobacteria , Ultraviolet Rays , Ultraviolet Rays/adverse effects , Ecosystem , Amino Acids/metabolism , Cyanobacteria/metabolism
14.
Molecules ; 28(14)2023 Jul 22.
Article in English | MEDLINE | ID: mdl-37513460

ABSTRACT

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.


Subject(s)
Amino Acids , Aquatic Organisms , Humans , Amino Acids/chemistry , Tissue Distribution , Aquatic Organisms/metabolism , Ultraviolet Rays
15.
Mar Drugs ; 21(6)2023 Jun 10.
Article in English | MEDLINE | ID: mdl-37367682

ABSTRACT

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.


Subject(s)
Lichens , Rhodophyta , Seaweed , Seaweed/chemistry , Lichens/chemistry , Countercurrent Distribution , Amino Acids/chemistry , Rhodophyta/chemistry , Plant Extracts/metabolism , Ultraviolet Rays
16.
Metab Eng ; 78: 137-147, 2023 07.
Article in English | MEDLINE | ID: mdl-37257683

ABSTRACT

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.


Subject(s)
Cyanobacteria , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Sunscreening Agents/chemistry , Sunscreening Agents/metabolism , Glycine/metabolism , Amino Acids/metabolism , Cyanobacteria/metabolism , Threonine , Serine/metabolism
17.
Foods ; 12(7)2023 Mar 30.
Article in English | MEDLINE | ID: mdl-37048293

ABSTRACT

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.

18.
Mar Drugs ; 21(4)2023 Apr 19.
Article in English | MEDLINE | ID: mdl-37103392

ABSTRACT

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.


Subject(s)
Amino Acids , Skin Neoplasms , Humans , Amino Acids/chemistry , Ultraviolet Rays/adverse effects , Sunscreening Agents/chemistry , Skin , Skin Neoplasms/prevention & control
19.
J Sci Food Agric ; 103(10): 5087-5095, 2023 Aug 15.
Article in English | MEDLINE | ID: mdl-36991224

ABSTRACT

BACKGROUND: Mycosporine-like amino acids (MAAs) are known as the strongest solar guardians in nature. RESULTS: 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. CONCLUSION: 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.


Subject(s)
Fishes , Animals , Gelatin/chemistry , Amino Acids/chemistry , Ultraviolet Rays , Starch/chemistry , Ascorbic Acid/chemistry
20.
Molecules ; 28(3)2023 Feb 02.
Article in English | MEDLINE | ID: mdl-36771087

ABSTRACT

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.


Subject(s)
Amino Acids , Cyanobacteria , Amino Acids/chemistry , Brazil , Cyanobacteria/genetics , Cyanobacteria/metabolism , Ultraviolet Rays , Multigene Family
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