Tetrodotoxin (TTX) is a potent marine neurotoxin responsible for severe seafood poisoning in humans, characterized by neurological symptoms that may be fatal. Originally identified as a natural contaminant of pufferfish (Tetraodontidae family), over the last few years TTX and its analogs have also been detected in other edible marine organisms, including mollusks, gastropods and crustaceans. Consequently, there is a need for rapid, sensitive, and reliable methods for TTX detection in seafood. In this study, a functional assay based on the use of mouse neuroblastoma Neuro-2a cells has been optimized and characterized for TTX detection in mussels. The assay is based on the toxin’s ability to block voltage-gated sodium channels, thereby counteracting the sodium-dependent cytotoxicity induced by veratridine and ouabain. The linear range of the TTX standard curve fell between 0.44 and 33 ng/mL, with limits of TTX detection (LOD) and quantitation (LOQ) of 0.132 ng/mL and 0.439 ng/mL, respectively, and good intra- and inter-day repeatability (RSDr= 15 and 11%, respectively). The assay also detected saxitoxin, which shares the same mechanism of action as TTX, but was less sensitive towards 4,9-anhydro-TTX. The minimum mussel extract dilution of 1:100 did not result in matrix-related interference, allowing accurate TTX quantitation, with a LOQ of 0.54 µg TTX equivalents/kg mussel meat. Given its sensitivity, the optimized Neuro-2a assay represents a promising tool for toxicity-based TTX quantitation in mussels before their consumption.
This study characterizes the genomic identity, functional efficacy, and computational biophysics of FNL62-AMP, a novel antimicrobial peptide isolated from a phylogenomically distinct, newly identified Brevibacillus species. Production kinetics revealed a late-exponential phase onset of antibacterial activity with sustained potency against methicillin-resistant Staphylococcus aureus (MRSA). LC-MS/MS analysis identified the peptide sequence as NH2-LLLLFR-COOH. FNL62-AMP demonstrated excellent formulation resilience, retaining full anti-MRSA activity under high thermal stress (80 °C for 6 h) and showing robust resistance to generic trypsin and proteinase K proteolysis. Formulative co-incubation assays demonstrated charge-dependent compatibility, where nonionic Triton X-100 preserved baseline efficacy while ionic surfactants induced antagonism. In vitro time-kill kinetics, scanning electron microscopy, and SYTOX Green assays confirmed rapid, concentration-dependent bactericidal action driven by immediate membrane permeabilization. Molecular dynamics simulations successfully captured the spontaneous self-assembly of 64 FNL62-AMP monomers into a stable macro-aggregate. This consolidation process was quantitatively characterized by a simultaneous contraction in the radius of gyration (Rg), a sharp drop in solvent-accessible surface area (SASA), and a transitional plateau in mean squared displacement (MSD). Ultimately, the high thermal stability, structural resilience, and predictable surfactant compatibility of FNL62-AMP propose ways to be developed for lead optimization and druggability.
Chemotherapy-induced neurotoxicity, affecting both the central and peripheral nervous systems, is a frequent and severe adverse effect of paclitaxel (PAC) treatment with limited therapeutic options. We previously demonstrated that PAC triggers neuronal cell death via ferroptosis. αO-Conotoxin GeXIVA[1,2], a marine-derived peptide, has shown efficacy in alleviating chemotherapy-induced neuropathic pain. In the present study, we investigated whether GeXIVA[1,2] protects neurons from PAC-induced neurotoxicity by suppressing ferroptosis. Using SH-SY5Y and HT-22 neuronal cell lines, we found that GeXIVA[1,2] pretreatment rescued PAC-impaired cell viability without exhibiting cytotoxicity. GeXIVA[1,2] markedly attenuated PAC-induced reactive oxygen species (ROS) overproduction and restored intracellular glutathione (GSH) levels. Mechanistically, PAC suppressed the Nrf2/SLC7A11/GSH/GPX4 ferroptosis-defense pathway, and GeXIVA[1,2] reactivated this axis by upregulating Nrf2, SLC7A11, and GPX4 protein expression. These results reveal a novel ferroptosis-suppressive function of GeXIVA[1,2] in the context of PAC-induced neurotoxicity. Our findings provide a mechanistic foundation for developing GeXIVA[1,2] as a ferroptosis-targeted intervention against chemotherapy-induced neurotoxicity.
Spodoptera litura is a polyphagous lepidopteran pest with rapidly evolving insecticide resistance. Peptide toxins from venomous organisms, many stabilized by the inhibitor cystine knot motif, offer bioinsecticidal potential but show limited oral efficacy. Seven previously reported candidates were recombinantly produced in Pichia pastoris and screened against S. litura by injection and leaf-disc feeding bioassays, followed by whole-plant spray assays. The marine nemertean toxin nemertide α-1, designated A1, showed the strongest oral activity, with a median lethal concentration of 3.97 μg μL−1. Conversely, the engineered spider-venom peptide U1-AGTX-Ta1b R9Q (T1) was most toxic by injection but showed no detectable oral activity, demonstrating that injection toxicity did not predict oral efficacy. High-cell-density fermentation yielded an estimated A1 titer of 1.20 g L−1. Whole-plant foliar application of A1 showed concentration-dependent efficacy. Mortality reached 92.07% at 8 μg μL−1, whereas lower concentrations suppressed larval growth, reducing the mean weight of surviving larvae by 41.55% and 67.58% at 1 and 2 μg μL−1, respectively, and mitigated feeding damage. These findings extend the documented oral insecticidal spectrum of nemertide α-1 to S. litura and demonstrate whole-plant efficacy following foliar application, supporting its development as an active ingredient for foliar-applied peptide bioinsecticides.
A fraction from an extract of the marine sponge Geodia microspinosa was identified as active in a high-throughput screen for molecules that impair the viability of diffuse pleural mesothelioma (DPM) cell lines. Bioassay-guided isolation led to the identification of microspinosamides B and D, and a new artifact, microspinosamide C, together with two previously reported analogues, microspinosamide and polydiscamide B, as the active principles. Their planar structures were solved by NMR and HRESIMS analyses, while advanced Marfey’s reaction, ROESY, and ECD were used for the establishment of their absolute configurations. All pure metabolites demonstrated low micromolar potency for the reduction in viability of the DPM cell lines.
The global seafood industry generates substantial quantities of processing by-products such as heads, viscera, skin, bones, scales, and shells. These residues represent an underutilized resource rich in proteins, lipids, minerals, enzymes, and polysaccharides. Conventional valorization approaches such as chemical extraction, wet rendering, and enzymatic hydrolysis have been used to recover valuable compounds from seafood waste. However, conventional methods often involve high chemical consumption, long processing times, and environmental concerns. Green extraction technologies have emerged as promising alternatives, with subcritical water extraction (SWE) gaining significant attention due to its unique properties and ability to simultaneously extract and convert biomass components. This review provides a comprehensive overview of the valorization of seafood processing wastes using SWE. Particular emphasis is placed on the physicochemical properties of subcritical water, the reaction mechanisms governing the hydrolysis and transformation of proteins, lipids, and polysaccharides, and the key parameters influencing extraction performance. Recent advances in the recovery of value-added products such as amino acids, bioactive peptides, protein hydrolysates, omega-3-rich oils, chitin derivatives, and mineral-rich materials are summarized. In addition, the integration of SWE with complementary technologies such as supercritical CO2 extraction, enzymatic hydrolysis, and hydrothermal carbonization is examined as a strategy for developing integrated seafood biorefineries.
The Potiguar Basin, in Rio Grande do Norte (Northeastern Brazil), has an extensive coastline with a rich diversity of marine sponges that remain poorly explored for their biotechnological potential. In this study, three sponge species—Suberites aurantiacus, Mycale (Zygomycale) angulosa, and Halichondrida—were collected, and a preliminary extraction was then performed to remove the most hydrophobic fraction with n-hexane. The remaining sediment was re-extracted with chloroform and methanol (1:1), and the resulting extracts (ESUB, EMYC, and EHALI, respectively) were analyzed. Chemical characterization using Gas–Liquid Chromatography–Mass Spectrometry (GC/MS) revealed distinct profiles for each genus, with the presence of alcohols, alkaloids, phenolic compounds, and lipids, mainly sterols. ESUB presented 17 identified compounds, EMYC had 15 identified and EHALI showed 15 identified. The antioxidant potential of the extracts was evaluated in vitro using DPPH radical scavenging, copper ion chelation, and total antioxidant capacity (TAC). The EC50 values (mg/mL) for DPPH were 0.05 (ESUB), 0.14 (EMYC), and 0.06 (EHALI), while for copper chelation they were 0.03, 0.034, and 0.026, respectively. Preliminary toxicological assays indicated low toxicity, with IC50 values above 9.6 mg/mL in human erythrocytes and Tenebrio molitor larvae, demonstrating a favorable safety profile for potential therapeutic applications. The study presents a preliminary screening of underexplored species in an equally underexplored region and supports further studies on their bioactive potential.
The growing demand for natural products in the pharmaceutical, nutraceutical, cosmetic, food, and feed sectors has stimulated considerable interest in the identification and sustainable production of bioactive compounds from renewable biological resources [...]
The search for antioxidant compounds from aquatic or marine environments to cope with the harmful effects of reactive oxygen species (ROS) is one of the major challenges in contemporary blue biotechnology. Among marine microorganisms, microalgae have emerged as promising candidates for the discovery and production of natural bioactive molecules with antioxidant properties. This review highlights the potential of microalgae and cyanobacteria as a sustainable source of antioxidant compounds and examines their growing relevance in biotechnology and pharmaceutical applications. A broad range of antioxidant metabolites produced by microalgae, including carotenoids, fatty acids, vitamins, polyphenols, and flavonoids, has also been discussed, with particular emphasis on their antioxidant mechanisms and bioactive properties. This review also integrates antioxidant mechanisms with the physiological and metabolic responses, underlying antioxidant production and sustainable strategies used to enhance their accumulation. Furthermore, microalgae offer the advantage of sustainable production systems, with the potential to enhance the biosynthesis and accumulation of valuable compounds through optimized cultivation strategies. Thus, different sustainable approaches aimed at increasing antioxidant compound production or reducing operational costs in microalgae cultivation are discussed to identify efficient and economically viable processes that maximize the biotechnological potential of these microorganisms for future industrial applications.
Although flaviviruses, including DENV, ZIKV and JEV, remain important causes of febrile, congenital, and neurological diseases, treatment options remain largely supportive, with limited availability of virus-specific antiviral therapies. Marine organisms and marine-derived microorganisms produce chemically distinct antiviral materials, including sulfated polysaccharides, terpenoids, alkaloids, peptides, cyclodepsipeptides, and polyketides. However, their activities range from preliminary extract-level inhibition to direct biochemical target validation, making mechanistic comparison difficult. This review critically evaluates marine-derived anti-flaviviral agents using two complementary dimensions, the infection stage implicated by experimental assays and the strength of evidence supporting that assignment. DENV evidence is dominated by sulfated algal macromolecules that interfere with adsorption or internalization, whereas ZIKV studies encompass lipophilic algal metabolites, fungal alkaloids, cyclodepsipeptides, and a few target-oriented candidates. Across the field, most reports remain stage-associated rather than target-validated. Cross-study potency comparisons are constrained by differences in virus strains, cell models, assay formats, and treatment schedules. JEV-specific evidence is particularly sparse. Based on the DENV and ZIKV evidence map, we propose concise priorities for JEV-oriented discovery: early compound-level dereplication, parallel cytotoxicity testing, orthogonal confirmation of productive infection, stage-resolved assays, and biochemical or genetic validation of conserved flaviviral targets. This evidence-based framework can help distinguish promising chemical candidate scaffolds from preliminary antiviral signals and guide mechanism-informed development of marine-derived natural products against flaviviruses.
Marine-derived rare actinomycetes are a chemically prolific yet underexploited source of structurally diverse secondary metabolites. In this review, rare actinomycetes are operationally defined as marine-derived non-Streptomyces actinomycetes that remain comparatively underexplored yet possess demonstrated or predicted capacity for specialized-metabolite biosynthesis. Genome sequencing has revealed that their biosynthetic potential greatly exceeds the range of metabolites recovered under standard cultivation conditions. However, many reported compounds remain only loosely associated with the gene clusters that encode them. This review provides a biosynthesis-centered perspective on marine-derived rare actinomycetes, focusing on secondary metabolites for which biosynthetic gene clusters (BGCs) or pathways have been proposed, experimentally assessed, or functionally validated. It focuses on compounds reported after 2017, along with earlier metabolites whose biosynthetic origins were resolved only later. Representative examples are organized by genus and structural class and weighed according to the level of evidence linking each metabolite to its BGC, ranging from bioinformatic prediction and metabolomic correlation to validation by gene inactivation, heterologous expression, and enzymatic characterization. The surveyed metabolites include polyketides, nonribosomal peptides, polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS) hybrids, siderophores, angucyclines, anthracyclines, macrolides, diketopiperazine derivatives, and other unusual scaffolds. Collectively, these findings indicate how integrating genome mining, metabolomics, and molecular networking with targeted biosynthetic experiments can accelerate marine natural product discovery and unravel novel enzymatic functions and biosynthetic mechanisms in rare actinomycetes.
This study evaluated the synergistic effects of sodium chloride (NaCl) and cytidine monophosphate (CMP) on enhancing the co-production of astaxanthin and fatty acids while suppressing secondary cell wall (SCW) formation in Haematococcus lacustris (synonym: H. pluvialis) under high-light stress. An orthogonal design identified the optimal combination (0.5 g/L NaCl and 0.5 mM CMP), which significantly increased astaxanthin yield by over 35.6% and total fatty acid yield by 28%, while maintaining 96.8% of cells in motile state (SCW-deficient). Physiological analyses revealed elevated reactive oxygen species levels, concomitant with higher actual photochemical efficiency (Fv′/Fm′) and relative electron transport rates II (rETR(II)) along with enhanced non-photochemical quenching (NPQ) capacity, and metabolic reprogramming characterized by the accumulation of lipids, sugars, and starch alongside decreased protein yield. Metabolomics indicated reduced carbon supply for SCW polysaccharide biosynthesis, coupled with decreased protein yield and altered amino acid profiles characteristic of nitrogen-limited metabolism, which collectively favored the reallocation of carbon resources toward nitrogen-free high-value products. Transcriptomics confirmed the downregulation of SCW component biosynthetic genes and the upregulation of the methylerythritol phosphate (MEP) pathway and astaxanthin biosynthetic pathway. Scale-up experiments validated this strategy for producing astaxanthin-rich motile cells, offering a promising approach for microalgal biorefinery.
Microalgae are a rich source of bioactive metabolites, including unsaturated fatty acids, chlorophylls, carotenoids, and phenolic compounds with antioxidant and nutraceutical potential. Salicylic acid (SA) and its volatile ester, methyl salicylate (MS), are well-known elicitors of secondary metabolism in higher plants, yet their mechanisms of action in green microalgae, particularly the effects of MS, remain poorly understood. Therefore, we evaluated how salicylates applied at concentrations of 0.1, 1, and 10 µM affect the growth and accumulation of bioactive compounds in a high-productivity clone of Planktochlorella nurekis, combining gas chromatography coupled mass spectrometry (GC-MS) fatty acid profiling with spectrophotometric determination of pigment levels, total phenols, and L-phenylalanine ammonia-lyase (PAL) activity. SA increased cell number and size, whereas 0.1 µM MS resulted in a six-fold gain in cell number. Accordingly, 10 µM MS raised chlorophyll b levels—nearly three-fold (from 76 to 211 µg per g D.W.)—and total carotenoids—roughly four-fold (129 to 520 µg per g D.W.)—while both elicitors significantly increased the monounsaturated fatty acid fraction (from 29% to about 40%). Notably, 10 µM SA lowered the levels of saturated fatty acids. Moreover, 10 µM MS doubled the PAL activity and increased the total phenols by 17% compared to the control. Thus, SA and MS act as effective elicitors of high-value metabolites in P. nurekis.
Fucoxanthin is a high-value marine xanthophyll with a unique epoxy-allene structure, predominantly produced by brown macroalgae and marine diatoms, and additionally reported in other heterokont lineages including chrysophytes and certain haptophytes. As a core light-harvesting pigment, it exhibits multiple bioactivities including lipid-lowering, antioxidant, and anti-inflammatory effects, with broad applications in functional foods, dietary supplements, and pharmaceuticals. Currently, over 70% of commercial fucoxanthin is extracted from brown macroalgae such as Saccharina japonica and Undaria pinnatifida. However, due to low endogenous pigment content and significant extraction losses, the industrial yield is only 0.05–0.1% of dry weight, failing to meet the growing downstream demand for stable high-purity supply. Marine diatoms, characterized by short growth cycles, high pigment content, and controllable culture conditions, have emerged as a core direction for industrial upgrading. Among them, Phaeodactylum tricornutum and Odontella aurita are the two most systematically studied high-yield strains with outstanding potential. This review adopts volumetric productivity (mg/(L·d)) as the core evaluation metric, which integrates biomass concentration, pigment content, and production cycle, and reflects industrial efficiency more reliably than single intracellular content data. We summarize high-yield induction methods from the dimensions of nutrient regulation, light optimization, exogenous induction, and strain improvement, and systematically compare downstream processing, safety profiles, and regulatory status between the two diatoms. Furthermore, we evaluate multi-product co-production potential and scale-up performance from a techno-economic perspective. This review provides a side-by-side benchmark of these two marine diatoms, offering a data-driven reference for process development and industrial deployment of microalgae-derived fucoxanthin.
Ulcerative colitis (UC) is a chronic inflammatory bowel disease with elusive etiology and limited therapeutic options, accompanied by potential side effects. In this study, to investigate the therapeutic potential of Antarctic krill oil (AKO) in a dextran sulfate sodium (DSS)-induced mouse colitis model, animal experiments, molecular assays, transcriptomics, metabolomics, and 16S rRNA gene sequencing were conducted. Our results revealed that supplementation of AKO significantly alleviated colitis symptoms, such as weight loss, and inflammatory responses. Moreover, multi-omics analyses demonstrated that AKO inhibited the PI3K/Akt signaling pathway, remodeled beneficial gut microbiota, and reshaped metabolite profiles associated with glycerolphospholipid metabolism. Remarkably, AKO alleviated DSS-induced colitis, accompanied by coordinated changes in the gut microbiota, metabolites, and transcriptome, which were associated with suppression of key inflammatory pathways. These findings present experimental evidence for the potential of AKO as a marine-based nutritional intervention for UC, and offer novel perspectives on microbiota-targeted therapies for inflammatory diseases.
Methicillin-resistant Staphylococcus aureus (MRSA) infections contribute significantly to the mortality rate associated with antimicrobial resistance, while also increasing medical complications when compared to methicillin-sensitive isolates. Consequently, increasing the efficacy of β-lactams may alleviate the burden associated with this drug–pathogen combination. Dialysed extracts, derived from Pelvetia canaliculata, were further partitioned to generate phlorotannin enriched extracts (PEEs). Minimum inhibitory concentrations (MICs) of extracts were used to determine the antimicrobial and antibiotic-modulating activity against seven clinical MRSA isolates. Whole-transcriptomic sequencing was carried out to determine potential mechanisms of action on the strongest extract–antibiotic combination against an MRSA isolate, determined by the fractional inhibitory concentration index (FICI). The most pronounced β-lactam modulatory effects were seen in the >100 kDa PEE from P. canaliculata in conjunction with amoxicillin (MIC fold-reductions = 12.8–214), while >30 kDa PEE resulted in the strongest FIC index value when combined with penicillin (0.23), indicating a synergistic effect. Whole-transcriptome analysis of this treatment identified multiple differentially expressed genes (Log2FC +1/−1) of statistical significance (p adj < 0.05), namely related to accelerated autolysis, cell-envelope stability, protein synthesis, DNA replication, iron homeostasis and active transport. This study highlights preliminary transcriptomic evidence of biological processes associated with penicillin modulation by P. canaliculata > 30 kDa PEE, providing a foundation for further investigation of its antibiotic-modulating activity.
Two new indole derivatives (1 and 2), along with fourteen known indole and polyketide compounds, were characterized from the Beibu Gulf coral-derived fungus Pestalotiopsis microspora GXIMD 02530. Their structures and absolute configurations were determined by comprehensive spectroscopic analysis, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction. Structurally, furoindolin A (1) was obtained as a rare racemic 6/5/5 tricyclic indole derivative incorporating a dihydrofuranone ring, which was further separated into a pair of enantiomers by chiral chromatographic resolution. Compounds (±)-1, 3, 4, and 6–12 exhibited inhibition of LPS-induced NF-κB luciferase activities. Phomopsilactone (10) displayed antibacterial activities against Staphylococcus epidermidis, Bacillus subtilis, and Staphylococcus aureus, with MIC values of 15.6, 31.25, and 62.5 μg/mL, respectively. Our findings would expand the chemical variety of indole derivatives and highlight furoindolin A (+)-1 as a promising chemical template for further biosynthetic and anti-inflammatory pharmacological investigation.
Fucoidan (FUC) exhibits immunomodulatory activity; however, its effects on intestinal mucosal immunity through dendritic cell (DC)-mediated regulation remain unclear. In this study, fucoidan was extracted from Saccharina japonica by hot-water extraction and characterized by chemical composition analysis, gel permeation chromatography (GPC), and Fourier-transform infrared spectroscopy (FT-IR). Bone marrow-derived DCs were used to evaluate the effects of FUC on DC maturation and immune function. An LPS-induced acute enteritis mouse model was used to assess intestinal injury, barrier function, and DC-mediated T/B cell immune responses. Structural analysis confirmed that purified FUC has the sulfated polysaccharide characteristics. FUC promoted DC maturation and enhanced antigen-presenting capacity. In LPS-induced enteritis, FUC reduced IL-1β, IL-6, and TNF-α levels and improved intestinal barrier integrity by restoring the mRNA expression of tight-junction-related genes. Mechanistically, FUC regulated the excessive activation of the TLR4/MyD88/NF-κB pathway, increased TGF-β and IFN-γ expression, modulated Th1/Th17/Treg immune balance, and promoted B cell homing and sIgA secretion. FUC regulates DC-mediated immune responses, repairs intestinal mucosal barrier function, and restores the intestinal immune microenvironment, thereby alleviating LPS-induced intestinal inflammation and maintaining intestinal homeostasis.
As a microalga of significant economic value, the polysaccharide synthesis efficiency and bioactivity of Spirulina platensis are significantly influenced by the culture condition. In this study, S. platensis H11 obtained by space mutagenesis, was systematically compared to the growth characteristics, polysaccharide properties, and antioxidant activities between seawater and freshwater culture conditions. The results showed that the biomass concentration (8.87 g L−1) and polysaccharide yield (5.46 g L−1) of the strain in the seawater condition were increased by 16.1% and 14.7%, respectively, compared with that in the freshwater condition. Seawater-derived polysaccharides (SSPS) featured an α-configuration glucan backbone and exhibited higher molecular weight (466.19 kDa) and an additional β-configured anomeric signal compared with FSPS. Despite having lower contents of sulfate groups (0.56% DW) and glucuronides (4.92% DW) by 39.3% and 33.5%, respectively, compared to FSPS (0.78% sulfate group, 6.57% glucuronide), SSPS exhibited significantly enhanced hydroxyl radical-scavenging activity, demonstrated by a decreased IC50 value of 2.53 mg mL−1 as opposed to 3.07 mg mL−1. The present study confirms that seawater cultivation is an effective strategy for inducing the synthesis of highly active polysaccharides in space mutant strains, and provides a theoretical basis for the targeted production of polysaccharides in S. platensis and their functional and precise application.
Sea anemone venom has attracted increasing attention in biomedical research due to its multifarious compounds with biological activities. Although the venom is predominantly made up of proteins, the diversity and complexity of these proteins remain poorly understood. In this work, the proteins derived from the tentacle and column of Heteractis magnifica were investigated by integrating transcriptomic and proteomic technologies. A total of 3573 protein sequences from transcriptome databases were identified and clustered into nine functional categories. We also performed proteomic analysis on the proteins identified in H. magnifica, and 339 proteins were found to be present in both datasets. Notably, a comprehensive analysis of six typical categories was implemented, and the representative proteins were explored in depth using multiple alignments, homology modeling and molecular docking. Meanwhile, a few low-copy but functionally intriguing proteins were discovered, highlighting the presence of unconventional components in sea anemone venom. This work provides the first holistic overview of the typical protein families and novel information on functional proteins from H. magnifica, contributing to a deeper understanding of sea anemone proteins and facilitating the discovery of potential proteins for marine drugs or biotechnological tools.
