In the present work, the marine bacterium Vibrio alginolyticus Mo245 demonstrated the ability to secrete an extracellular polysaccharide (EPS) concomitant with the intracellular accumulation of a polyhydroxyalkanoate (PHA). Upon cultivation in glycerol as the sole carbon source, V. alginolyticus Mo245 co-produced a glycosaminoglycan (GAG), named EPS-Mo245, and the homopolymer poly(3-hydroxybutyrate) (P3HB), attaining maximum concentrations of 1.29±0.01 g/L and 1.78±0.00 g/L, respectively, within a 24 h cultivation. EPS-Mo245 was a hyaluronic acid (HA)-like GAG, composed of glucuronic acid (40.5±2.2 mol

Biofouling is a major drawback in the marine industry. As the use of the toxic antifouling coatings has been restricted due to their negative environmental impact, the development of environmentally friendly, biocide-free alternative is necessary. Hydrogel coatings based on polysaccharides are an alternative that is attracting a great deal of attention. These coatings are considered a promising material because they are environmentally friendly and the dense hydration layer protects the substrate from biofouling. This study reports on optimized hydrogel coatings based on chitosan that improve their antifouling performance by incorporating a hydrophilic polysaccharide. This study compares the performance of a bacterial exopolysaccharide from Vibrio MO245 with that of the well-known hyaluronic acid. After presenting the preparation of the coatings, including the depolymerization of MO245, the coatings characterize using topography measurement, crosslinking analysis, hydration capacity analysis and hydrophobicity analysis. The efficiency of the coatings is then evaluated against three representative organisms: the bacterium Vibrio harveyi, the diatoms Cylindrotheca closterium and the larvae Magallana gigas.

Marine microorganisms produce extracellular polysaccharides with unique features, such as the presence of unusual monosaccharides, adhesiveness to biotic and abiotic surfaces, and close similarity to glycosaminoglycans (GAG), that are of considerable interest for biotechnological applications. In the present study, an extracellular microbial polysaccharide (EPS) has been isolated and structurally characterized. The EPS producer is the Gram-negative bacterium Alteromonas macleodii Mo169, previously isolated from a marine environment in French Polynesia. The bacterium has been cultivated using glycerol as the sole carbon source. Interestingly, the secreted EPS was composed of 2-acetamido-2-deoxy-guluronic acid (GulNA) and 2-acetamido-2-deoxy-glucose (GlcN), two amino sugars not commonly found in the Alteromonas genus. The purified EPS, with a molecular weight of 0.30 ± 0.01 MDa, consisted of the following repeating unit: →4)-α-L-GulNAc3OAcA-(1 → 3)-β-D-GlcNAc-(1 → .

The Zika virus (ZIKV), an emerging orthoflavivirus, presents a significant public health threat due to its rapid dissemination and association with severe neurological complications. The urgent need for effective antiviral agents has driven research into novel bioactive compounds derived from unique natural sources. Microorganisms inhabiting extreme environments are particularly promising for such discoveries due to their potential to produce unique metabolites. In this study, we explored microorganisms from the underexplored French Polynesian microbial mats known as "Kopara" to identify new bioactive natural products. Using a molecular networking-based dereplication strategy, we investigated various culture and extraction techniques of the strain Nocardia otitidiscaviarum 20-S13, leading to the discovery of two novel glycoglycerolipids, otitiglycomycins A and B (1 and 2). Structure elucidation of these compounds was achieved through NMR spectroscopy, X-ray crystallography, and TDDFT-specific rotation prediction. We found that otitiglycomycin A (1), but not otitiglycomycin B (2), suppresses ZIKV infection at non cytotoxic concentrations without effects on cell viability. Time-of-drug addition assays along with virus inactivation and binding assays demonstrated that 1 neutralizes ZIKV infectivity by preventing the virus from attaching to the host cell membrane.

Biological cryopreservation often involves using a cryoprotective agent (CPA) to mitigate lethal physical stressors cells endure during freezing and thawing, but effective CPA concentrations are cytotoxic. Hence, natural polysaccharides have been studied as biocompatible alternatives. Here, a subset of 26 natural polysaccharides of various chemical composition was probed for their potential in enhancing the metabolic post-thaw viability (PTV) of cryopreserved Vero cells. The best performing cryoprotective polysaccharides contained significant fucose amounts, resulting in average PTV 2.8-fold (up to 3.1-fold) compared to 0.8-fold and 2.2-fold for all non-cryoprotective and cryoprotective polysaccharides, respectively, outperforming the optimized commercial CryoStor™ CS5 formulation (2.6-fold). Stoichiometrically, a balance between fucose (18-35.7 mol%), uronic acids (UA) (13.5-26 mol%) and high molecular weight (MW > 1 MDa) generated optimal PTV. Principal component analysis (PCA) revealed that fucose enhances cell survival by a charge-independent, MW-scaling mechanism (PC1), drastically different from the charge-dominated ice growth disruption of UA (PC2). Its neutral nature and unique properties distinguishable from other neutral monomers suggest fucose may play a passive role in conformational adaptability of polysaccharide to ice growth inhibition, or an active role in cell membrane stabilization through binding. Ultimately, fucose-rich anionic polysaccharides may indulge in polymer-ice and polymer-cell interactions that actively disrupt ice and minimize lethal volumetric fluctuations due to a balanced hydrophobic-hydrophilic character. Our research showed the critical role neutral fucose plays in enhancing cellular cryopreservation outcomes, disputing previous assumptions of polyanionicity being the sole governing predictor of cryoprotection.

Recently, polysaccharide-based hydrogels crosslinked with the trivalent iron cation have attracted interest due to their remarkable properties that include high mechanical stability, stimuli-responsiveness, and enhanced absorptivity. In this study, a Fe3+ crosslinked hydrogel was prepared using the biocompatible extracellular polysaccharide (EPS) secreted by the marine bacterium Alteromonas macleodii Mo169. Hydrogels with mechanical strengths (G') ranging from 0.3 kPa to 44.5 kPa were obtained as a result of the combination of different Fe3+ (0.05-9.95 g L-1) and EPS (0.3-1.7 %) concentrations. All the hydrogels had a water content above 98 %. Three different hydrogels, named HA, HB, and HC, were chosen for further characterization. With strength values (G') of 3.2, 28.9, and 44.5 kPa, respectively, these hydrogels might meet the strength requirements for several specific applications. Their mechanical resistance increased as higher Fe3+ and polymer concentrations were used in their preparation (the compressive hardness increased from 8.7 to 192.1 kPa for hydrogel HA and HC, respectively). In addition, a tighter mesh was noticed for HC, which was correlated to its lower swelling ratio value compared to HA and HB. Overall, this preliminary study highlighted the potential of these hydrogels for tissue engineering, drug delivery, or wound healing applications.

Spirobistetrodecamycins A and B (1 and 2), tetrodecamycin-derived vinyl cyclobutane dimers, are herein reported for the first time. Their structures including absolute configurations were elucidated by extensive spectroscopic methods, single-crystal X-ray diffraction, and ECD calculations. As a way to provide a rationale for the spontaneous formation of these dimers, the irradiation of tetrodecamycin has been carried out leading to the production of regioisomeric [2 + 2] head-to-head 1 and head-to-tail 2. These two compounds showed no antibacterial activity against Escherichia coli, Micrococcus luteus, and Bacillus subtilis.

In this study, the novel exopolysaccharide produced by the marine isolate Alteromonas macleodii Mo 169 was used as stabilizing and reducing agent for the synthesis of silver and gold nanoparticles (AgNP and AuNP, respectively). Both the synthesized NP and the EPS/NP composites were characterized. The synthesized nanoparticles were spherical with an average particle size of 16 and 15 nm for AuNP and AgNP, respectively. The cytotoxicity of the EPS and the bio-nanocomposites was evaluated on human keratinocyte (HaCaT) and fibroblast (CCD-1079Sk) cell lines. None of the samples demonstrated cytotoxicity for concentrations up to 1000 mg L-1. In vitro wound healing experiments demonstrated that the bio-nanocomposites EPS/AgNP and EPS/AuNP (100 mg L-1) promoted cell migration, thus contributing for a faster wound recovery. These findings demonstrated for the first time the potential of A. macleodii Mo169 EPS to synthesize and stabilize nanoparticles useful for the development of novel biomaterials with application in the biomedical field.

A sugar-rich apple pulp waste generated from fruit processing for juice production was used as the sole carbon source for the cultivation of Alteromonas macleodii Mo169, a marine bacterium known for its EPS-secreting ability. The strain efficiently utilized the glucose and fructose present in the apple pulp waste, reaching biomass and EPS production of 9.20 +/- 0.61 and 3.51 +/- 0.08 g L-1, respectively, in 24-hour bioreactor cultivation. Two high molecular weight (Mw) fractions (1.7 +/- 0.0 and 0.74 +/- 0.0 MDa) were detected in the sample recovered from the cell-free supernatant by dialysis. The compositional analysis revealed the presence of glucose (31.1 +/- 0.2 mol%), arabinose (23.9 +/- 0.1 mol%), mannose (17.3 +/- 0.1 mol%), glucosamine (10.3 +/- 0.5 mol%), galactose (8.7 +/- 0.0 mol%) and galacturonic acid (8.7 +/- 0.0 mol%), as well as a high content in sulphate (6.0 +/- 0.5 wt%). Given the presence of a high Mw polysaccharide in the apple pulp waste, probably pectin, a fraction of the detected sugar monomers might be attributed to that polymer, which was recovered together with A. macleodii Mo169 EPS. Concomitant with EPS synthesis, there was a viscosity build-up in the cultivation broth, which developed a shear-thinning fluid behaviour not observed in the initial medium. Therefore, this study demonstrates that apple pulp waste can be efficiently converted into a novel polysaccharide by A. macleodii Mo169 in a sustainable bioprocess. Moreover, the EPS sugar and acyl composition, together with its good thickening capacity, render the biopolymer of interest for use in several applications. (c) 2023 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc- nd/4.0/).

In this study, the novel exopolysaccharide (EPS) produced by the marine bacterium Alteromonas macleodii Mo 169 was used as a stabilizer and capping agent in the preparation of selenium nanoparticles (SeNPs). The synthesized nanoparticles were well dispersed and spherical with an average particle size of 32 nm. The cytotoxicity of the EPS and the EPS/SeNPs bio-nanocomposite was investigated on human keratinocyte (HaCaT) and fibroblast (CCD-1079Sk) cell lines. No cytotoxicity was found for the EPS alone for concentrations up to 1 g L-1. A cytotoxic effect was only noticed for the bio-nanocomposite at the highest concentrations tested (0.5 and 1 g L-1). In vitro experiments demonstrated that non-cytotoxic concentrations of the EPS/SeNPs bio-nanocomposite had a significant cellular antioxidant effect on the HaCaT cell line by reducing ROS levels up to 33.8%. These findings demonstrated that the A. macleodii Mo 169 EPS can be efficiently used as a stabilizer and surface coating to produce a SeNP-based bio-nanocomposite with improved antioxidant activity.

Due to environmental regulations, antifouling marine coatings must be gradually replaced by biocide-free coatings. Marine organisms weakly adhere to fouling release coatings, presenting a low surface free energy and a high elasticity, so they can be readily removed by the sheer force of water. Currently, these materials are mainly composed of petrochemical polymers, such as silicone or fluoropolymers, with hydrophilic polymers as additives. However, following the ever-increasing environmental concerns, the research on new, alternative, eco-friendly coatings is oriented towards the use of biobased polymers from renewable resources. Two main families have been studied: polyhydroxyalkanoates (PHAs) and polysaccharides. PHAs are produced by bacteria in stressful conditions, while polysaccharides are extracted from plants, animals, or micro-organisms such as bacteria, in which case they are called exopolysaccharides (EPS). Since the use of these polymers is a non-toxic approach to controlling fouling colonization, this review provides an overview of these biobased polymers for their applications in new anti-adhesive marine coatings.

Biofilms, responsible for many serious drawbacks in the medical and marine environment, can grow on abiotic and biotic surfaces. Commercial anti-biofilm solutions, based on the use of biocides, are available but their use increases the risk of antibiotic resistance and environmental pollution in marine industries. There is an urgent need to work on the development of ecofriendly solutions, formulated without biocidal agents, that rely on the anti-adhesive physico-chemical properties of their materials. In this context, exopolysaccharides (EPSs) are natural biopolymers with complex properties than may be used as anti-adhesive agents. This study is focused on the effect of the EPS MO245, a hyaluronic acid-like polysaccharide, on the growth, adhesion, biofilm maturation, and dispersion of two pathogenic model strains, Pseudomonas aeruginosa sp. PaO1 and Vibrio harveyi DSM19623. Our results demonstrated that MO245 may limit biofilm formation, with a biofilm inhibition between 20 and 50%, without any biocidal activity. Since EPSs have no significant impact on the bacterial motility and quorum sensing factors, our results indicate that physico-chemical interactions between the bacteria and the surfaces are modified due to the presence of an adsorbed EPS layer acting as a non-adsorbing layer.

This work assessed the film-forming capacity of exopolysaccharides (EPS) produced by six Alteromonas strains recently isolated from different marine environments in French Polynesia atolls. The films were transparent and resulted in small colour alterations when applied over a coloured surface (ΔEab below 12.6 in the five different colours tested). Moreover, scanning electron microscopy showed that the EPS films were dense and compact, with a smooth surface. High water vapour permeabilities were observed (2.7–6.1 × 10−11 mol m−1 s−1 Pa−1), which are characteristic of hydrophilic polysaccharide films. The films were also characterised in terms of barrier properties to oxygen and carbon dioxide. Interestingly, different behaviours in terms of their mechanical properties under tensile tests were observed: three of the EPS films were ductile with high elongation at break (ε) (35.6–47.0%), low tensile strength at break (Ꞇ) (4.55–11.7 MPa) and low Young’s modulus (εm) (10–93 MPa), whereas the other three were stiffer and more resistant with a higher Ꞇ (16.6–23.6 MPa), lower ε (2.80–5.58%), and higher εm (597–1100 MPa). These properties demonstrate the potential of Alteromonas sp. EPS films to be applied in different areas such as biomedicine, pharmaceuticals, or food packaging.

Rheology modifiers are essential additives in numerous products in a variety of industries. Due to environmental awareness, consumer-oriented industries are interested in novel natural rheological agents that can replace synthetic chemicals. In this study, the chemical composition and rheological properties of a novel exopolysaccharide (EPS) produced by Alteromonas macleodii Mo 169 were investigated. It was mainly composed of uronic acids (50 mol%) and total carbohydrates were 17 % sulfated. The EPS viscosity increased with concentration, and a non-Newtonian shear thinning behavior was found for concentrations above 0.1 wt%. The elastic and viscous moduli indicated a weak gel-like structure above 0.4 wt%. It maintained its shear thinning behavior and viscoelastic properties in the presence of NaCl and CaCl2 for pH range 5-7 and temperatures up to 55 degrees C. Though the apparent viscosity decreased at pH 3 and 9 and temperatures above 65 degrees C, the shear thinning behavior was retained. The viscous and viscoelastic properties were recovered after heating (95 degrees C) and cooling (0 degrees C), indicating a good thermal stability and recoverability. After high shear force, the solution recovered original rheological properties within few seconds, demonstrating self-healing properties.

Solid-phase extraction embedded dialysis (SPEED technology) is an innovative procedure developed to physically separate in-situ, during the cultivation, the mycelium of filament forming microorganisms, such as actinomycetes and fungi, and the XAD-16 resin used to trap the secreted specialized metabolites. SPEED consists of an external nylon cloth and an internal dialysis tube containing the XAD resin. The dialysis barrier selects the molecular weight of the trapped compounds, and prevents the aggregation of biomass or macromolecules on the XAD beads. The external nylon promotes the formation of a microbial biofilm, making SPEED a biofilm supported cultivation process. SPEED technology was applied to the marine Streptomyces albidoflavus 19-S21, isolated from a core of a submerged Kopara sampled at 20 m from the border of a saltwater pond. The chemical space of this strain was investigated effectively using a dereplication strategy based on molecular networking and in-depth chemical analysis. The results highlight the impact of culture support on the molecular profile of Streptomyces albidoflavus 19-S21 secondary metabolites.

Marine environments comprise almost three quarters of Earth’s surface, representing the largest ecosystem of our planet. The vast ecological and metabolic diversity found in marine microorganisms suggest that these marine resources have a huge potential as sources of novel commercially appealing biomolecules, such as exopolysaccharides (EPS). Six Alteromonas strains from different marine environments in French Polynesia atolls were selected for EPS extraction. All the EPS were heteropolysaccharides composed of different monomers, including neutral monosaccharides (glucose, galactose, and mannose, rhamnose and fucose), and uronic acids (glucuronic acid and galacturonic acid), which accounted for up to 45.5 mol% of the EPS compositions. Non-carbohydrate substituents, such as acetyl (0.5–2.1 wt%), pyruvyl (0.2–4.9 wt%), succinyl (1–1.8 wt%), and sulfate (1.98–3.43 wt%); and few peptides (1.72–6.77 wt%) were also detected. Thermal analysis demonstrated that the EPS had a degradation temperature above 260 °C, and high char yields (32–53%). Studies on EPS functional properties revealed that they produce viscous aqueous solutions with a shear thinning behavior and could form strong gels in two distinct ways: by the addition of Fe2+, or in the presence of Mg2+, Cu2+, or Ca2+ under alkaline conditions. Thus, these EPS could be versatile materials for different applications.

MO245 exopolysaccharide (EPS) was produced in laboratory conditions from Vibrio genus microorganism isolated from bacterial mats found in Moorea Island. Its structure consists of a linear tetrasaccharide repeating unit →4)-β-D-GlcpA-(1→4)-α-D-GalpNAc-(1→3)-β-D-GlcpNAc-(1→4)-β-D-GlcpA-(1→ containing covalently-linked 5% of glucose, galactose, and rhamnose, determined by methylation analyses and NMR spectroscopy. The molecular weight, radius of gyration (Rg) and intrinsic viscosity, [η], determined by gel permeation chromatography with light scattering and viscosity detection, were 513 ± 4 kDa (PDI, 1.42 ± 0.01), 6.7 ± 0.3 dl/g and 56 ± 0.3 nm respectively. The chelation of the EPS with copper divalent ions leads to the instantaneous formation of gels. The structural similitude proposed, based in an equal ratio of GlcA to N-acetylated sugars and in the same type of glyosidic linkages present in the repeating unit (alternated 1→3 and 1→4 linkages), is translated into analogous physicochemical properties: MO245 EPS is a flexible polyelectrolyte, with scaling exponents similar to that described for HA. This similitude opens opportunities in future drug delivery, tissue engineering, and cosmetic applications.

A library of microorganisms originating from various marine environments in French Polynesia was screened for polyhydroxyalkanoate producing bacteria. No significant connection was found between the geo-ecological source of bacteria and their ability to produce polyhydroxyalkanoate. A bacterial strain designated as Enterobacter FAK 1384 was isolated from a shark jaw. When grown on coprah oil, this bacterium produces a PHA constituting of 62 mol % 3-hydroxydecanoate and lower amount of 12 mol % 3-hydroxydodecenoate and of 7.6 mol % 3-hydroxydodecanoate. These interesting properties make this mcl-PHA a good candidate for further exploitations in many industrial sectors, as in film and coating manufacturing, as well as for biomedical applications.

It is well known that microorganisms well-adapted to survival in extreme ecosystems could be considered as new sources of biomolecules that have biotechnological importance. On French Polynesian atolls, microbial mats are developing in water ponds exposed to fluctuations in physical and chemical parameters. In these microbial mats, which are called "kopara" by the inhabitants, bacteria coexist with cyanobacteria, and a synergistic relationship may exist between these two types of living microorganisms. A large number of cyanobacteria and bacteria have been isolated from different mats. Under laboratory conditions, these microorganisms were shown to produce various exopolymers, including exopolysaccharides and poly-beta-hydroxyalkanoates, along with pigments for further commercial developments. This manuscript gives an overview of substances isolated and characterized from these bacteria and cyanobacteria and discusses their potential applications in biotechnology. (C) 2010 Elsevier Ltd. All rights reserved.