Ulva rigida Valorization into Poly(3-hydroxybutyrate), Organic Acids and Functional Ingredients.

Halomonas elongata 1H9T is a moderate halophilic strain able to produce poly(3-hydroxybutyrate) (P(3HB)), a biodegradable plastic, and gluconic acid, a valuable organic acid with wide industrial applications. In this work, the green alga Ulva rigida was used as platform to produce cultivation substrates for microbial conversion as well as functional ingredients, targeting its full valorization. The liquor obtained by autohydrolysis presented the highest concentration of oligosaccharides and protein, being an interesting feedstock to produce functional ingredients. The acid and/or enzymatic hydrolysis liquors are adequate as substrates for microbial processes. Shake flask assays with H. elongata revealed that the N-rich liquor produced after acidic treatment was the best suited for cell growth while the N-poor liquor produced by the enzymatic treatment of acid-pretreated algae residues produced the highest P(3HB) titers of 4.4 g/L. These hydrolysates were used in fed-batch cultivations as carbon and protein sources for the co-production of gluconic acid and polymer achieving titers of 123.2 g/L and 7.2 g/L, respectively. Besides gluconic acid, the Krebs cycle intermediate 2-oxoglutaric acid, also called alpha-ketoglutaric acid (KGA), was produced. Therefore, the co-production of P(3HB) and acids may be of considerable interest as an algal biorefinery valorization strategy.

Biochemical characterization of a new ulvan lyase and its applicability in utilization of ulvan and preparation of ulva oligosaccharides.

Ulva is globally distributed specie and has a high economic value. Ulvan is one of the main active substances in Ulva, which has a variety of biological properties. Ulvan lyase degrades ulvan through a ß-elimination mechanism which cleaves the ß-glycosidic bond between Rha3S and GlcA or IdoA. The complex monosaccharide composition of ulvan makes it promising for use in food and pharmaceutical applications. This thesis explores a putative ulvan lyase from Alteromonas sp. KUL_42. We expressed and purified the protein, performed a series of characterizations and signal peptide had been removed. The results showed that the protein molecular weight of ULA-2 was 53.97 kDa, and it had the highest catalytic activity at 45 °C and pH 8.0 in Tris-HCl buffer. The Km and Vmax values were 2.24 mg · mL-1 and 2.048 µmol · min-1 · mL-1, respectively. The activity of ULA-2 was able to maintain more than 80% at 20 ~ 30 °C. ESI-MS analysis showed that the primary end-products were mainly disaccharides to tetrasaccharides. The study of ULA-2 enriches the ulvan lyase library, promotes the development and high-value utilization of Ulva resources, and facilitates further research applications of ulvan lyase in ulva oligosaccharides.

Airborne dimethyl sulfide (DMS) cues dimethylsulfoniopropionate (DMSP) increases in the intertidal green alga Ulva fenestrata.

Although the use of airborne molecules as infochemicals is common in terrestrial plants, it has not been shown to occur in an ecologically relevant context in marine seaweeds. Like terrestrial plants, intertidal plants spend part of their lives emersed at low tide and release volatile organic compounds (VOCs) into the air when they are grazed or physiologically stressed. We hypothesized seaweeds could use airborne VOCs as infochemicals and respond to them by upregulating a keystone defensive metabolite, dimethylsulfoniopropionate (DMSP). We conducted laboratory and field experiments in which Ulva fenestrata was exposed to airborne dimethyl sulfide (DMS), a volatile antiherbivore and antioxidant metabolite released when the seaweed is grazed or physiologically stressed. In the laboratory, U. fenestrata exposed to DMS had 43-48% higher DMSP concentrations, relative to controls, 6-9 days after exposure. In the field, U. fenestrata 1 m downwind of DMS emitters had 19% higher DMSP concentrations than upwind seaweeds after 11 days. To our knowledge, this is the first demonstration of a marine plant using an airborne molecule released when damaged to elicit defensive responses. Our study suggests that the ability to detect airborne compounds has evolved multiple times or before the divergence of terrestrial plants and green algae.

Regulatory mechanisms of the edible alga Ulva lactuca polysaccharide via modulation of gut microbiota in diabetic mice.

In this study, Ulva lactuca polysaccharide (ULP) antihyperglycemic effect was assessed by monitoring changes in the gut microbiota of aging diabetic mice. The results showed that ULP alleviated type 2 diabetes by improving insulin tolerance, increasing SOD and CAT activities, and thus lowering blood glucose level. Moreover, ULP regulated the expressions of INSR and AMPK concurrent with inhibition the expression of JNK, JAK, STAT3, p16 and p38 to improve glucose metabolism dysfunction. Interestingly, the abundance of Alloprevotella and Pediococcus change might the key factor for ULP antihyperglycemic effectiveness in aging-related diabetes. These results suggest that ULP can exert a mechanism of blood glucose regulation by improving intestinal diversity composition asides from direct insulin mimetic actions.

Activated abscisic acid pathway and C4 pathway, inhibited cell cycle progression, responses of Ulva prolifera to short term high temperature elucidated by multi-omics.

The annual outbreak of green tides since 2007 has destroyed coastal waters' ecological environment and caused substantial economic losses. Ulva prolifera, known as the dominant species of green tides, is influenced by temperatures. Omics-based technology was used to analyze U. prolifera under 12 h of treatment at 30 °C in the work. High temperature has the following advantages, e.g., activating the abscisic acid signaling pathway, improving the heat tolerance of U. prolifera, up-regulating metabolites such as glycolipids, glyceroyl, and glutamic acid to maintain the stability and fluidity of cells, and reducing the stimulatory effect of external stress on cells. The key genes and proteins of the tricarboxylic acid cycle, glycolysis, and pentose phosphorylation pathways were inhibited; however, the key enzyme pyruvate phospho-dikinase of the C4 pathway was up-regulated. The C4 pathway was activated in U. prolifera in response to high-temperature stress and may play a key role in photosynthesis. Besides, U. prolifera metabolizing amino acids was active. High temperature inhibited genes and proteins related to DNA replication and cell cycle in the transcriptome and proteome as well as the growth and reproduction of U. prolifera.

Purification identification and function analysis of ACE inhibitory peptide from Ulva prolifera protein.

In the present study, Ulva prolifera, an edible alga, was used to prepare angiotensin-I converting enzyme (ACE) inhibitory peptide. The algae protein was isolated and later hydrolyzed by five commercial enzymes (alcalase, papain, pepsin, trypsin, neutral protease), either individually or in combination. Hydrolysate, with the highest in vitro ACE inhibitory activity, was processed using the Sephadex-G100, ultrafiltration, HPLC-Q-TOF-MS, ADMET screening and molecular docking, respectively. The ACE inhibitory peptide DIGGL with a IC50 value of 10.32 ± 0.96 µM was then identified. The peptide against ACE by a non-competitive mode and mainly attributable to the three Conventional Hydrogen Bonds. It could activate Endothelial nitric oxide synthase activity in NO generation and reduce Endothelin-1 secretion induced by Angiotensin II in Human umbilical vein endothelial cells. Meanwhile, DIGGL could promote mice splenocytes proliferation, which was also effective when co-incubated with Con A or LPS, respectively. Besides, the anti-ACE peptide could remain active during the digestion of gastrointestinal proteases (pepsin-trypsin) in vitro.

From model organism to application: Bacteria-induced growth and development of the green seaweed Ulva and the potential of microbe leveraging in algal aquaculture.

The marine green macroalga Ulva (Chlorophyta, Ulvales), also known as sea lettuce, coexists with a diverse microbiome. Many Ulva species proliferate in nature and form green algal blooms ("green tides"), which can occur when nutrient-rich wastewater from agricultural or densely populated areas is flushed into the sea. Bacteria are necessary for the adhesion of Ulva to its substrate, its growth, and the development of its blade morphology. In the absence of certain bacteria, Ulva mutabilis develops into a callus-like morphotype. However, with the addition of the necessary marine bacteria, the entire morphogenesis can be restored. Surprisingly, just two bacteria isolated from U. mutabilis are sufficient for inducing morphogenesis and establishing the reductionist system of a tripartite community. While one bacterial strain causes algal blade cell division, another causes the differentiation of basal cells into a rhizoid and supports cell wall formation because of a low concentration of the morphogen thallusin (below 10-10 mol/L). This review focuses on the research conducted on this topic since 2015, discusses how U. mutabilis has developed into a model organism in chemical ecology, and explores the questions that have already been addressed and the perspectives that a reductionist model system allows. In particular, the field of systems biology will achieve a comprehensive, quantitative understanding of the dynamic interactions between Ulva and its associated bacteria to better predict the behavior of the system as a whole. The reductionist approach has enabled the study of the bacteria-induced morphogenesis of Ulva. Specific questions regarding the optimization of cultivation conditions as well as the yield of raw materials for the food and animal feed industries can be answered in the laboratory and through applied science. Genome sequencing, the improvement of genetic engineering tools, and the first promising attempts to leverage macroalgae-microbe interactions in aquaculture make this model organism, which has a comparatively short parthenogenetic life cycle, attractive for both fundamental and applied research. The reviewed research paves the way for the synthetic biology of macroalgae-associated microbiomes in sustainable aquacultures.

Stereoselective Total Synthesis of (-)-Thallusin for Bioactivity Profiling.

Chemical mediators are key compounds for controlling symbiotic interactions in the environment. Here, we disclose a fully stereoselective total synthesis of the algae differentiation factor (-)-thallusin that utilizes sophisticated 6-endo-cyclization chemistry and effective late-stage sp2 -sp2 -couplings using non-toxic reagents. An EC50 of 4.8 pM was determined by quantitative phenotype profiling in the green seaweed Ulva mutabilis (Chlorophyte), underscoring this potent mediator's enormous, pan-species bioactivity produced by symbiotic bacteria. SAR investigations indicate that (-)-thallusin triggers at least two different pathways in Ulva that may be separated by chemical editing of the mediator compound structure.

Differential responses of bloom-forming Ulva intestinalis and economically important Gracilariopsis lemaneiformis to marine heatwaves under changing nitrate conditions.

Marine heatwaves (MHWs) are affecting the survival of macroalgae. However, little is known regarding how the impacts of MHWs are regulated by nitrogen availability. In this study, we investigated the physiological and genetic responses of a green-tide macroalga Ulva intestinalis Linnaeus and a commercially cultivated macroalga Gracilariopsis lemaneiformis (Bory) E.Y. Dawson, Acleto & Foldvik under different nitrate conditions to simulated MHWs. Under nitrogen limited conditions (LN), heatwaves did not significantly affect biomass or Fv/Fm of U. intestinalis although it led to an earlier biomass decline due to more reproduction events, and meanwhile an upregulation in genes related to TCA cycle and oxidative phosphorylation was detected, supporting sporulation. Under nitrogen replete conditions (HN), heatwaves did not change biomass, Fv/Fm or photosynthetic pigments but reduced reproduction rate along with insignificant change of oxidative phosphorylation and TCA cycle related genes. Meanwhile, genes related to photosynthesis and glutathione metabolism were upregulated. Regarding G. lemaneiformis, heatwaves reduced its Fv/Fm and photosynthetic pigments content, leading to bleaching and death, and photosynthesis-related genes were also downregulated at LN. Fv/Fm was improved and photosynthesis-related genes were up-regulated by the combination of nitrogen enrichment and heatwaves, whereas G. lemaneiformis remained bleached and died by day 12. Therefore, U. intestinalis could survive heatwaves through shifting to micropropagules at LN and protecting its photosynthesis at HN. In contrast, G. lemaneiformis died of bleaching when suffering heatwaves regardless of nitrogen availability. These findings suggest that in future oceans with eutrophication and MHWs, the harmful alga U. intestinalis may have more advantages over the economic alga G. lemaneiformis.

Changes in dissolved organic pool and regulation of associated nutrients during green tides: A case study of Ulva prolifera bloom in the southern Yellow Sea.

To explore the spatial and temporal distributions of dissolved organic matter (DOM) affected by Ulva prolifera bloom and the regulation of DOM released by the addition of nitrate (NO3-N) nutrients, the variation in DOM was analyzed by three cruises in the coastal waters of the southern Yellow Sea (SYS) off Qingdao during a 2018 U. prolifera bloom. A mesocosm experiment was conducted during the decay of U. prolifera. The U. prolifera bloom resulted in higher concentrations of the bulk DOM except for total dissolved amino acids (TDAA), compared with levels recorded during non-bloom periods. Three different fluorescent substances of chromophoric dissolved organic matter (CDOM) were identified, which were also affected by the bloom. Concentrations of total dissolved carbohydrates (TCHO), monosaccharides (MCHO), and polysaccharides (PCHO) peaked during the late bloom, with average levels of 49.78 ± 9.73, 14.23 ± 3.32, and 35.55 ± 9.19 µmol C L-1, respectively, accompanied by an increase of DOC, indicating release of DOM with U. prolifera decay. In contrast, the concentrations of CDOM and TDAA declined during the late bloom, with averages of 0.10 ± 0.10 m-1 and 1.16 ± 0.82 µmol L-1, suggesting that the yields of CDOM and TDAA were regulated by the U. prolifera bloom and other mechanism. These findings were confirmed by the mesocosm experiment. DOM production was obviously improved by NO3-N addition except for TDAA and CDOM. The release of protein-like CDOM was primarily regulated by NO3-N nutrients, however, it was not obvious with humic-like CDOM. Our findings provide evidence supporting the role of U. prolifera blooms in the coastal DOM pool and its significant effect on the marine carbon cycle.

Balancing Damage via Non-Photochemical Quenching, Phenolic Compounds and Photorespiration in Ulva prolifera Induced by Low-Dose and Short-Term UV-B Radiation.

The Yellow Sea green tide (YSGT) is the world's largest transregional macroalgal blooms, and the causative species Ulva prolifera (U. prolifera) suffers from ultraviolet-b radiation (UVBR) during the floating migration process. Previous study confirmed that U. prolifera displayed a wide variety of physiological responses characterized as acclimation to UVBR, while the response mechanisms against low-dose and short-term radiation (LDSTR) are not clear. A study with photosynthetically active radiation (PAR) and UVBR was designed: normal light (NL: 72 µmol photons m-2 s-1), NL+0.3 (UVBR: 0.3 W·m-2), and NL+1.6 (UVBR: 1.6 W·m-2). The results showed that high-dose UVBR inhibited photosynthesis in thalli, especially under long-term exposure, while a variety of physiological responses were observed under LDSTR. The inhibition of photosynthesis appeared to be ameliorated by the algae under LDSTR. Further analysis showed that U. prolifera achieved balancing damage by means of non-photochemical quenching (NPQ), accumulation of phenolic compounds coupled with the ASA-GSH cycle involved in the antioxidant process and enhanced photorespiratory metabolism under LDSTR. This study provides new insights into the balancing damage mechanisms of U. prolifera under LDSTR, enabling the thalli to adapt to the light conditions during the long duration and distance involved in floating migration.

Seaweed Extracts: A Promising Source of Antibiofilm Agents with Distinct Mechanisms of Action against Pseudomonas aeruginosa.

The organization of bacteria in biofilms is one of the adaptive resistance mechanisms providing increased protection against conventional treatments. Thus, the search for new antibiofilm agents for medical purposes, especially of natural origin, is currently the object of much attention. The objective of the study presented here was to explore the potential of extracts derived from three seaweeds: the green Ulva lactuca, the brown Stypocaulon scoparium, and the red Pterocladiella capillacea, in terms of their antibiofilm activity against P. aeruginosa. After preparation of extracts by successive maceration in various solvents, their antibiofilm activity was evaluated on biofilm formation and on mature biofilms. Their inhibition and eradication abilities were determined using two complementary methods: crystal violet staining and quantification of adherent bacteria. The effect of active extracts on biofilm morphology was also investigated by epifluorescence microscopy. Results revealed a promising antibiofilm activity of two extracts (cyclohexane and ethyl acetate) derived from the green alga by exhibiting a distinct mechanism of action, which was supported by microscopic analyses. The ethyl acetate extract was further explored for its interaction with tobramycin and colistin. Interestingly, this extract showed a promising synergistic effect with tobramycin. First analyses of the chemical composition of extracts by GC-MS allowed for the identification of several molecules. Their implication in the interesting antibiofilm activity is discussed. These findings suggest the ability of the green alga U. lactuca to offer a promising source of bioactive candidates that could have both a preventive and a curative effect in the treatment of biofilms.

Metal-free carbocatalysts derived from macroalga biomass (Ulva lactuca) for the activation of peroxymonosulfate toward the remediation of polycyclic aromatic hydrocarbons laden marine sediments and its impacts on microbial community.

Potential toxic chemicals, specifically, polycyclic aromatic hydrocarbons (PAHs), are major sediment contaminants. Herein, green seaweed (Ulva lactuca) was used as a feedstock and pyrolyzed at temperature in the range between 300 and 900 °C. The metal-free carbocatalyst (GSBC) for peroxymonosulfate (PMS) activation to degrade PAHs contaminated sediments was studied. The effects of GSBC‒PMS treatment on microbial community abundance was studied as well. The pyrolysis temperature of GSBC preparation affected the PMS activation performance. Results show that GSBC700 exhibited remarkable catalytic characteristics in PAHs degradation by effective activation of PMS. The results also demonstrated that the sulfate radical-carbon-driven advanced oxidation processes (SR-CAOP) reaction achieved 87% and apparent rate constant (kobs) of 6.3 × 10-2 h-1 of total PAHs degradation in 24 h at 3.3 g/L of GSBC, PMS dose of 1 × 10-4 M, and pH 3.0. The degradation of 2-, 3-, 4-, 5-, and 6-ring PAHs was 84, 83, 83, 80, and 89%, respectively. The synergetic effect established between GSBC and PMS enhanced the formation of ROSs, namely, SO4-, HO, and 1O2, which were major species contributing to PAHs degradation. The synergistic effect of π‒π stacking structure and graphitization of GSBC formed electron shuttle, which contributed to PAHs degradation performance. Microbial community structure analyses in the GSBC‒PMS treated sediments showed that the relative abundance of Lactobacillus_rhamnosus species, most of which belonged to the Lactobacillus genus and Firmicutes phylum, which aided in continuing PAHs biodegradation post GSBC‒PMS treatment. Therefore, GSBC can be a promising carbocatalyst produced via biomass-to-biochar conversion as biowaste-to-energy source used in the SR-CAOP-mediated process for sediment remediation.

Polyphenols extracted from Enteromorpha clathrata alleviates inflammation in lipopolysaccharide-induced RAW 264.7 cells by inhibiting the MAPKs/NF-κB signaling pathways.

ETHNOPHARMACOLOGY RELEVANCE: Enteromorpha has long been recorded in traditional Chinese medicine, with cholesterol-lowering, anti-cancer, anti-inflammatory and antibacterial effects. Recently, we extracted the polyphenol-enriched fraction from Enteromorpha clathrata (E. clathrata) by ethyl acetate (ECPs), and isolated six individual polyphenols from ECPs via high-speed counter-current chromatography (HSCCC) with high-performance liquid chromatography (HPLC). AIM OF THE STUDY: In this study, we explored the anti-inflammatory activity and underlying mechanism of ECPs in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. MATERIALS AND METHODS: ECPs and the six polyphenols were used for nitric oxide (NO) assay to identify the components with potent inflammation inhibitory effect. Enzyme-linked immunosorbent assay (ELISA), quantitative real-time PCR (qPCR), flow cytometry, and Western blot analysis were applied to further investigate their anti-inflammatory effects and underlying mechanism in LPS-stimulated RAW264.7 cells. RESULTS: ECPs and the three individual polyphenols, including (-)-epicatechin, epigallocatechin-3-O-gallate and (-)-epicatechin-3-O-gallate, showed in vitro immunosuppressive activity by altering the cell biology at the gene, protein and functional levels in a dose- and species-dependent manner. Their anti-inflammatory effects were achieved by inhibiting LPS-induced production of nitric oxide and its upstream enzyme inducible nitric oxide synthase (iNOS), the pro-inflammatory cytokines including interleukin-1 beta (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), as well as the phagocytotic capacity, without cytotoxicity. The mechanism study further revealed that these anti-inflammatory properties were, at least partly, attributed to the suppressed activation of nuclear factor-κB (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signaling pathways. CONCLUSIONS: These findings indicated for the first time the correlation between the anti-inflammatory activity of ECPs and NF-κB and MAPK signaling pathways, suggesting that polyphenol-enriched organic fraction of E. clathrata could be potential candidate as therapeutic agent for treating inflammatory diseases.

Structural characterization of ulvans extracted from blade (Ulva ohnoi) and filamentous (Ulva tepida and Ulva prolifera) species of cultivated Ulva.

Ulvans from Ulva ohnoi, Ulva tepida and Ulva prolifera were extracted under mild acidic conditions, isolated and their composition and structure determined. The ulvans contained mostly rhamnose (31.6-46.7 mol%) and glucuronic acid (26.6-37.5 mol%), with smaller amounts of xylose (3.4-10.4 mol%) and iduronic acid (3.1-7.6 mol%). In addition, the ulvan samples also contained galactose (4.4-26.0 mol%). Glycosyl linkage analysis showed that ulvan from U. ohnoi contained mostly →4)-GlcpA-(1→ and →3,4)-Rhap-(1→. Preparation of partially methylated alditol acetate standards of idose showed that U. ohnoi contained →4)-IdopA-(1→. In addition to these residues, glycosyl linkage analysis of U. tepida and U. prolifera showed the presence of →2,3,4)-Rhap-(1→, →4)-Xylp-(1→, →2,4)-GlcpA-(1→ and →3,4)-GlcpA-(1→. These two species also contained galactose linkages. These data, together with nuclear magnetic resonance (NMR) spectroscopy indicated that U. ohnoi comprised mostly of type A3S ulvanobiuronic acid repeats [→4)-ß-D-GlcpA-(1→4)-α-L-Rhap3S-(1→], together with smaller amounts of type B3S ulvanobiuronic acid repeats [→4)-α-L-IdopA-(1→4)-α-L-Rhap3S-(1→] and ulvanobiose (U3S [→4)-ß-D-Xylp-(1→4)-α-L-Rhap3S-(1→]). NMR spectra of U. tepida and U. prolifera showed resonances not detected in U. ohnoi, highlighting the complexity of the ulvans from these species. Regardless of the structural diversity of the ulvan samples there was very little antioxidant or inhibitory activity detected on enzymatic processes investigated.

Ulva compressa from Copper-Polluted Sites Exhibits Intracellular Copper Accumulation, Increased Expression of Metallothioneins and Copper-Containing Nanoparticles in Chloroplasts.

In order to analyze the mechanisms involved in copper accumulation in Ulva compressa, algae were collected at control sites of central and northern Chile, and at two copper-polluted sites of northern Chile. The level of intracellular copper, reduced glutathione (GSH), phytochelatins (PCs), PC2 and PC4, and transcripts encoding metallothioneins (MTs) of U. compressa, UcMT1, UcMT2 and UcMT3, were determined. Algae of control sites contained around 20 µg of copper g-1 of dry tissue (DT) whereas algae of copper-polluted sites contained 260 and 272 µg of copper g-1 of DT. Algae of control sites and copper-polluted sites did not show detectable amounts of GSH, the level of PC2 did not change among sites whereas PC4 was increased in one of the copper-polluted sites. The level of transcripts of UcMT1 and UcMT2 were increased in algae of copper-polluted sites, but the level of UcMT3 did not change. Algae of a control site and a copper-polluted site were visualized by transmission electron microscopy (TEM) and the existence of copper in electrodense particles was analyzed using energy dispersive x-ray spectroscopy (EDXS). Algae of copper-polluted sites showed electrodense nanoparticles containing copper in the chloroplasts, whereas algae of control sites did not. Algae of a control site, Cachagua, were cultivated without copper (control) and with 10 µM copper for 5 days and they were analyzed by TEM-EDXS. Algae cultivated with copper showed copper-containing nanoparticles in the chloroplast whereas control algae did not. Thus, U. compressa from copper-polluted sites exhibits intracellular copper accumulation, an increase in the level of PC4 and expression of UcMTs, and the accumulation of copper-containing particles in chloroplasts.

The non-sulfated ulvanobiuronic acid of ulvans is the smallest active unit able to induce an oxidative burst in dicot cells.

Ulvans from green algae are promising compounds for plant protection because they are environmentally friendly and induce plant defense responses. We analyzed the structure-function relationship of ulvan polymers and oligomers for their elicitor activity in suspension-cultured cells of three dicot species. The polysaccharide from Ulva fasciata was characterized regarding its monosaccharide composition, degree of sulfation, and molecular mass. The polymer was partially depolymerized using acid hydrolysis, and the oligomers were separated using size exclusion chromatography. The oligomeric fractions were analyzed revealing mostly sulfated and de-sulfated ulvan dimers. Both the polymer and the oligomer fractions induced an NADPH oxidase-dependent oxidative burst in plant cells. The elicitor activity of the ulvan dimers did not require sulfation. By identifying the smallest elicitor-active unit, HexA-Rha, we took an important next step to understand how the structure influences ulvan elicitor responses. The desulfated ulvan dimer is discussed as a promising agro-biologic for sustainable agriculture.

Multi-scale modeling of intensive macroalgae cultivation and marine nitrogen sequestration.

Multi-scale macroalgae growth models are required for the efficient design of sustainable, economically viable, and environmentally safe farms. Here, we develop a multi-scale model for Ulva sp. macroalgae growth and nitrogen sequestration in an intensive cultivation farm, regulated by temperature, light, and nutrients. The model incorporates a range of scales by incorporating spatial effects in two steps: light extinction at the reactor scale (1 m) and nutrient absorption at the farm scale (1 km). The model was validated on real data from an experimental reactor installed in the sea. Biomass production rates, chemical compositions, and nitrogen removal were simulated under different seasons, levels of dilution in the environment and water-exchange rate in the reactor. This multi-scale model provides an important tool for environmental authorities and seaweed farmers who desire to upscale to large bioremediation and/or macroalgae biomass production farms, thus promoting the marine sustainable development and the macroalgae-based bioeconomy.

24(R, S)-Saringosterol - From artefact to a biological medical agent.

Enhancing the cholesterol turnover in the brain via activation of liver x receptors can restore memory in a mouse model for Alzheimer's disease. The edible Asian brown alga Sargassum fusiforme (Hijiki) contains high amounts of oxysterols such as (3ß, 24ξ)-stigmasta-5, 28-dien-3, 24-diol (24[R, S]-saringosterol) that are a potent liver x receptor agonists. We aimed to find native European seaweed species with contents of 24(R, S)-saringosterol that are comparable to those found in Sargassum fusiforme. Additionally, we hypothesize that seasonal variations modify the amount of 24(R, S)-saringosterol in seaweeds. Sterols and oxysterols were extracted with chloroform/methanol from various seaweed species harvested in the Eastern Scheldt in different seasons between October 2016 and September 2017. Identification and quantification of the lipids was performed by gas chromatography- mass spectrometry and gas chromatography- flame ionization detection. We confirmed that brown algae Undaria pinnatifida harvested in February and Sargassum muticum harvested in October contained the highest amounts of 24(R, S)-saringosterol (32.4 ± 15.25 µg/g, mean ± S.D. and 32.95 ± 2.91 µg/g, respectively) and its precursor fucosterol (1.48 ± 0.11 mg/g), higher than Sargassum fusiforme (20.94 ± 3.00 µg/g, mean ± S.D.), while Ascophyllum nodosum and Fucus vesiculosus and Fucus serratus contained amounts of 24(R, S)-saringosterol (22.09 ± 3.45 µg/g, 18.04 ± 0.52 µg/g and 19.47 ± 9.01 µg/g, mean ± S.D., respectively) comparable to Sargassum fusiforme. In other algae only minor amounts of these sterols were observed. The green algae Ulva lactuca contained only 0.29 mg/g fucosterol and 10.3 µg/g 24 (R, S)-saringosterol, while all investigated red algae did not contain any 24(R, S)-saringosterol or fucosterol. In the Eastern Scheldt algae harvested in September/October delivered the highest yield for 24(R, S)-saringosterol, with the exception of Undaria pinnatifida that showed the highest levels in February. We showed that exposure of lipid extracts of Ulva lactuca to sunlight at room temperature or in the presence of oxygen to UV-C light lead to the quantitative conversion of fucosterol into 24(R, S)-saringosterol. Exposing pure fucosterol to UV-light did not convert any fucosterol into 24(R, S)-saringosterol underscoring the requirement of seaweed constituents in the conversion of fucosterol into 24(R, S)-saringosterol. In conclusion, we showed that brown seaweeds harvested from the Eastern Scheldt contain amounts of 24(R, S)-saringosterol comparable to Sargassum fusiforme, varying per season and showing the highest amounts in spring. In accordance with these observations the amount of 24(R, S)-saringosterol in the brown seaweeds can be modulated by light.

Seaweed cellulose scaffolds derived from green macroalgae for tissue engineering.

Extracellular matrix (ECM) provides structural support for cell growth, attachments and proliferation, which greatly impact cell fate. Marine macroalgae species Ulva sp. and Cladophora sp. were selected for their structural variations, porous and fibrous respectively, and evaluated as alternative ECM candidates. Decellularization-recellularization approach was used to fabricate seaweed cellulose-based scaffolds for in-vitro mammalian cell growth. Both scaffolds were confirmed nontoxic to fibroblasts, indicated by high viability for up to 40 days in culture. Each seaweed cellulose structure demonstrated distinct impact on cell behavior and proliferation rates. The Cladophora sp. scaffold promoted elongated cells spreading along its fibers' axis, and a gradual linear cell growth, while the Ulva sp. porous surface, facilitated rapid cell growth in all directions, reaching saturation at week 3. As such, seaweed-cellulose is an environmentally, biocompatible novel biomaterial, with structural variations that hold a great potential for diverse biomedical applications, while promoting aquaculture and ecological agenda.