Sustainable production of heavy metal-binding levan by a subarctic permafrost thaw lake Pseudomonas strain 2ASCA.

Pseudomonas strain 2ASCA isolated in subarctic Québec, Canada, produced a cell membrane bound levan-type exopolymer (yield 1.17 g/L), after incubation in growth media containing 6 % sucrose (w/v) at temperature of 15 °C for 96 h. The objective of this study was to optimize levan production by varying the growth parameters. Moreover, the polymer's chemical characterization has been studied with the aim of increasing knowledge and leading to future applications in many fields, including heavy metal remediation. Higher levan yields (7.37 g/L) were reached by setting up microbial fermentation conditions based on the re-use of the molasses obtained from sugar beet processing. Spectroscopy analyses confirmed the levan-type nature of the exopolymer released by strain 2ASCA, consisting of a ß-(2,6)-linked fructose repeating unit. Gel permeation chromatography revealed that the polymer has a molecular weight of 13 MDa. Scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS) showed that the levan sequestered with a strong affinity Cr(III), which has never been previously reported, highlighting an interesting biosorption potential. In addition, SEM analysis revealed the formation of nanoparticles in acidified water solution.

Physico-Chemical Properties and Valorization of Biopolymers Derived from Food Processing Waste.

The widespread use of synthetic plastics, as well as the waste produced at the end of their life cycle, poses serious environmental issues. In this context, bio-based plastics, i.e., natural polymers produced from renewable resources, represent a promising alternative to petroleum-based materials. One potential source of biopolymers is waste from the food industry, the use of which also provides a sustainable and eco-friendly solution to waste management. Thus, the aim of this work concerns the extraction of polysaccharide fractions from lemon, tomato and fennel waste. Characterizing the chemical-physical and thermodynamic properties of these polysaccharides is an essential step in evaluating their potential applications. Hence, the solubility of the extracted polysaccharides in different solvents, including water and organic solvents, was determined since it is an important parameter that determines their properties and applications. Also, acid-base titration was carried out, along with thermoanalytical tests through differential scanning calorimetry. Finally, the electrospinning of waste polysaccharides was investigated to explore the feasibility of obtaining polysaccharide-based membranes. Indeed, electrospun fibers are a promising structure/system via which it is possible to apply waste polysaccharides in packaging or well-being applications. Thanks to processing feasibility, it is possible to electrospin waste polysaccharides by combining them with different materials to obtain porous 3D membranes made of nanosized fibers.

Inhibition of Enzymes Involved in Neurodegenerative Disorders and Aß1-40 Aggregation by Citrus limon Peel Polyphenol Extract.

Alzheimer's (AD) and Parkinson's diseases (PD) are multifactorial neurogenerative disorders of the Central Nervous System causing severe cognitive and motor deficits in elderly people. Because treatment of AD and PD by synthetic drugs alleviates the symptoms often inducing side effects, many studies have aimed to find neuroprotective properties of diet polyphenols, compounds known to act on different cell signaling pathways. In this article, we analyzed the effect of polyphenols obtained from the agro-food industry waste of Citrus limon peel (LPE) on key enzymes of cholinergic and aminergic neurotransmission, such as butyryl cholinesterase (BuChE) and monoamine oxidases (MAO)-A/B, on Aß1-40 aggregation and on superoxide dismutase (SOD) 1/2 that affect oxidative stress. In our in vitro assays, LPE acts as an enzyme inhibitor on BuChE (IC50 ~ 73 µM), MAO-A/B (IC50 ~ 80 µM), SOD 1/2 (IC50 ~ 10-20 µM) and interferes with Aß1-40 peptide aggregation (IC50 ~ 170 µM). These results demonstrate that LPE behaves as a multitargeting agent against key factors of AD and PD by inhibiting to various extents BuChE, MAOs, and SODs and reducing Aß-fibril aggregation. Therefore, LPE is a promising candidate for the prevention and management of AD and PD symptoms in combination with pharmacological therapies.

Extremophilic Microorganisms for the Green Synthesis of Antibacterial Nanoparticles.

The biogenic synthesis of nanomaterials, i.e., synthesis carried out by means of living organisms, is an emerging technique in nanotechnology since it represents a greener and more eco-friendly method for the production of nanomaterials. In this line, in order to find new biological entities capable of biogenic synthesis, we tested the ability of some extremophilic microorganisms to carry out the biogenic production of AgNPs and SeNPs. Silver NPs were produced extracellularly by means of the thermophilic Thermus thermophilus strain SAMU; the haloalkaliphilic Halomonas campaniensis strain 5AG was instead found to be useful for the synthesis of SeNPs. The structural characterization of the biogenic nanoparticles showed that both the Ag and Se NPs possessed a protein coating on their surface and that they were organized in aggregates. Moreover, both types of NPs were found be able to exert an interesting antibacterial effect against either Gram-positive or Gram-negative species. This study confirmed that extremophilic microorganisms can be considered valuable producers of biologically active nanoparticles; nevertheless, further experiments must be performed to improve the synthesis protocols in addition to the downstream processes.

Characterization of the exopolymer-producing Pseudoalteromonas sp. S8-8 from Antarctic sediment.

A synergistic approach using cultivation methods, chemical, and bioinformatic analyses was applied to explore the potential of Pseudoalteromonas sp. S8-8 in the production of extracellular polymeric substances (EPSs) and the possible physiological traits related to heavy metal and/or antibiotic resistance. The effects of different parameters (carbon source, carbon source concentration, temperature, pH and NaCl supplement) were tested to ensure the optimization of growth conditions for EPS production by the strain S8-8. The highest yield of EPS was obtained during growth in culture medium supplemented with glucose (final concentration 2%) and NaCl (final concentration 3%), at 15 °C and pH 7. The EPS was mainly composed of carbohydrates (35%), followed by proteins and uronic acids (2.5 and 2.77%, respectively) and showed a monosaccharidic composition of glucose: mannose: galactosamine: galactose in the relative molar proportions of 1:0.7:0.5:0.4, as showed by the HPAE-PAD analysis. The detection of specific molecular groups (sulfates and uronic acid content) supported the interesting properties of EPSs, i.e. the emulsifying and cryoprotective action, heavy metal chelation, with interesting implication in bioremediation and biomedical fields. The analysis of the genome allowed to identify a cluster of genes involved in cellulose biosynthesis, and two additional gene clusters putatively involved in EPS biosynthesis. KEY POINTS: • A cold-adapted Pseudoalteromonas strain was investigated for EPS production. • The EPS showed emulsifying, cryoprotective, and heavy metal chelation functions. • Three gene clusters putatively involved in EPS biosynthesis were evidenced by genomic insights.

Genomic Analysis Provides New Insights Into Biotechnological and Industrial Potential of Parageobacillus thermantarcticus M1.

Parageobacillus thermantarcticus strain M1 is a Gram-positive, motile, facultative anaerobic, spore forming, and thermophilic bacterium, isolated from geothermal soil of the crater of Mount Melbourne (74°22' S, 164°40' E) during the Italian Antarctic Expedition occurred in Austral summer 1986-1987. Strain M1 demonstrated great biotechnological and industrial potential owing to its ability to produce exopolysaccharides (EPSs), ethanol and thermostable extracellular enzymes, such as an xylanase and a ß-xylosidase, and intracellular ones, such as xylose/glucose isomerase and protease. Furthermore, recent studies revealed its high potential in green chemistry due to its use in residual biomass transformation/valorization and as an appropriate model for microbial astrobiology studies. In the present study, using a systems-based approach, genomic analysis of P. thermantarcticus M1 was carried out to enlighten its functional characteristics. The elucidation of whole-genome organization of this thermophilic cell factory increased our understanding of biological mechanisms and pathways, by providing valuable information on the essential genes related to the biosynthesis of nucleotide sugar precursors, monosaccharide unit assembly, as well as the production of EPSs and ethanol. In addition, gene prediction and genome annotation studies identified genes encoding xylanolytic enzymes that are required for the conversion of lignocellulosic materials to high-value added molecules. Our findings pointed out the significant potential of strain M1 in various biotechnological and industrial applications considering its capacity to produce EPSs, ethanol and thermostable enzymes via the utilization of lignocellulosic waste materials.

Effects of Ionizing Radiation and Long-Term Storage on Hydrated vs. Dried Cell Samples of Extremophilic Microorganisms.

A main factor hampering life in space is represented by high atomic number nuclei and energy (HZE) ions that constitute about 1% of the galactic cosmic rays. In the frame of the "STARLIFE" project, we accessed the Heavy Ion Medical Accelerator (HIMAC) facility of the National Institute of Radiological Sciences (NIRS) in Chiba, Japan. By means of this facility, the extremophilic species Haloterrigena hispanica and Parageobacillus thermantarcticus were irradiated with high LET ions (i.e., Fe, Ar, and He ions) at doses corresponding to long permanence in the space environment. The survivability of HZE-treated cells depended upon either the storage time and the hydration state during irradiation; indeed, dry samples were shown to be more resistant than hydrated ones. With particular regard to spores of the species P. thermantarcticus, they were the most resistant to irradiation in a water medium: an analysis of the changes in their biochemical fingerprinting during irradiation showed that, below the survivability threshold, the spores undergo to a germination-like process, while for higher doses, inactivation takes place as a consequence of the concomitant release of the core's content and a loss of integrity of the main cellular components. Overall, the results reported here suggest that the selected extremophilic microorganisms could serve as biological model for space simulation and/or real space condition exposure, since they showed good resistance to ionizing radiation exposure and were able to resume cellular growth after long-term storage.

A novel bio-solid phase extractor for preconcentrations of Hg and Sn in food samples.

An ecofriendly preconcentration method was developed based on the use of Geobacillus galactosidasius sp. nov immobilized on Amberlite XAD-4 as an adsorbent for the preconcentrations of Hg and Sn. SEM-EDX performed for the investigation of surface functionality and morphology. The detailed investigations of factors such as pH of the solution, flow rate, interfering ions and sample volume have been thanks to the optimization of the pre-concentration system. The optimum pHs were found as 5.0-7.0 for Hg and Sn and also the optimum flow rates were determined as 2 mL min-1 for recovery of Hg and Sn. Under the best experimental conditions, limits of detections (LOD) were found as 0.53 ng mL-1 for Hg and 0.27 ng mL-1 for Sn. RSDs were calculated as 8.2% for Hg and 6.9% for Sn. The process was validated to use certified references (fish samples). ICP-OES was used to measure the levels of Hg and Sn in various real meal patterns after the devised technique was used. Concentrations of Hg and Sn were quantitively measured on gluten-free biscuit, flour, rice, Tuna fish, meat, chicken meat, potato, chocolate, coffee, tap water, energy drink and mineral water samples with low RSD. The developed method emerges as an innovative technology that will eliminate the low cost and toxic effect.

Inhibition of Interleukin-6-Induced Matrix Metalloproteinase-2 Expression and Invasive Ability of Lemon Peel Polyphenol Extract in Human Primary Colon Cancer Cells.

Among matrix metalloproteinases (MMPs), MMP-9/2 are key enzymes involved in the proteolysis of extracellular matrices in the inflammatory process and in cancer. Since MMP-9/2 expression levels, activity, and secretion is up-regulated during inflammation in response to pro-inflammatory cytokines, such as interleukin-6 (IL-6), many efforts have been devoted to identifying factors that could inhibit the IL-6-induced MMP-9/2 expression. Up to now, several reports indicated that polyphenols from fruits and vegetables are among the major components of health promotion for their antioxidant properties and also for their anti-inflammatory and anti-cancer agents. Among plant derived polyphenols, lemon (Citrus limon) peel extract (LPE) shows anti-cancer properties in various cancer types. In our previous work, we demonstrated that LPE can reduce IL-6-induced migration/invasiveness and MMP-9/2 up-regulation in some gastric cancer cell lines. This study aims to exploit the anti-cancer properties of LPE using an in vitro system model of inflammation, consisting of IL-6-exposed human primary colon cancer cells. We first analyzed the effect of LPE on IL-6-induced cell migration and invasiveness by wound healing and Boyden chamber assay, respectively. The MMP-2 mRNA expression levels and gelatinolytic activity in the cell culture media were determined by q-PCR analysis and gelatin zymography, respectively, and finally, the effects of LPE on IL-6-induced JAK2/STAT3 signaling pathways have been investigated by Western blotting analysis. Our results show that LPE is able to inhibit the IL-6-dependent cell migration and invasiveness associated with the up-regulation of MMP-2 expression levels and that these effects are correlated to the STAT3 phosphorylation in human primary T88 and T93 colon cancer cells.

Effect of Carbon Sources in Carotenoid Production from Haloarcula sp. M1, Halolamina sp. M3 and Halorubrum sp. M5, Halophilic Archaea Isolated from Sonora Saltern, Mexico.

The isolation and molecular and chemo-taxonomic identification of seventeen halophilic archaea from the Santa Bárbara saltern, Sonora, México, were performed. Eight strains were selected based on pigmentation. Molecular identification revealed that the strains belonged to the Haloarcula, Halolamina and Halorubrum genera. Neutral lipids (quinones) were identified in all strains. Glycolipid S-DGD was found only in Halolamina sp. strain M3; polar phospholipids 2,3-O-phytanyl-sn-glycerol-1-phosphoryl-3-sn-glycerol (PG), 2,3-di-O-phytanyl-sn-glycero-1-phospho-3'-sn-glycerol-1'-methyl phosphate (PGP-Me) and sodium salt 1-(3-sn-phosphatidyl)-rac-glycerol were found in all the strains; and one unidentified glyco-phospholipid in strains M1, M3 and M4. Strains M1, M3 and M5 were selected for further studies based on carotenoid production. The effect of glucose and succinic and glutamic acid on carotenoid production was assessed. In particular, carotenoid production and growth significantly improved in the presence of glucose in strains Haloarcula sp. M1 and Halorubrum sp. M5 but not in Halolamina sp. M3. Glutamic and succinic acid had no effect on carotenoid production, and even was negative for Halorubrum sp. M5. Growth was increased by glutamic and succinic acid on Haloarcula sp. M1 but not in the other strains. This work describes for first time the presence of halophilic archaea in the Santa Bárbara saltern and highlights the differences in the effect of carbon sources on the growth and carotenoid production of haloarchaea.

Involvement of a Quorum Sensing Signal Molecule in the Extracellular Amylase Activity of the Thermophilic Anoxybacillus amylolyticus.

Anoxybacillus amylolyticus is a moderate thermophilic microorganism producing an exopolysaccharide and an extracellular α-amylase able to hydrolyze starch. The synthesis of several biomolecules is often regulated by a quorum sensing (QS) mechanism, a chemical cell-to-cell communication based on the production and diffusion of small molecules named "autoinducers", most of which belonging to the N-acyl homoserine lactones' (AHLs) family. There are few reports about this mechanism in extremophiles, in particular thermophiles. Here, we report the identification of a signal molecule, the N-butanoyl-homoserine lactone (C4-HSL), from the milieu of A. amylolyticus. Moreover, investigations performed by supplementing a known QS inhibitor, trans-cinnamaldehyde, or exogenous C4-HSL in the growth medium of A. amylolyticus suggested the involvement of QS signaling in the modulation of extracellular α-amylase activity. The data showed that the presence of the QS inhibitor trans-cinnamaldehyde in the medium decreased amylolytic activity, which, conversely, was increased by the effect of exogenous C4-HSL. Overall, these results represent the first evidence of the production of AHLs in thermophilic microorganisms, which could be responsible for a communication system regulating thermostable α-amylase activity.

Isolation and Characterization of Strain Exiguobacterium sp. KRL4, a Producer of Bioactive Secondary Metabolites from a Tibetan Glacier.

Extremophilic microorganisms represent a unique source of novel natural products. Among them, cold adapted bacteria and particularly alpine microorganisms are still underexplored. Here, we describe the isolation and characterization of a novel Gram-positive, aerobic rod-shaped alpine bacterium (KRL4), isolated from sediments from the Karuola glacier in Tibet, China. Complete phenotypic analysis was performed revealing the great adaptability of the strain to a wide range of temperatures (5-40 °C), pHs (5.5-8.5), and salinities (0-15% w/v NaCl). Genome sequencing identified KRL4 as a member of the placeholder genus Exiguobacterium_A and annotation revealed that only half of the protein-encoding genes (1522 of 3079) could be assigned a putative function. An analysis of the secondary metabolite clusters revealed the presence of two uncharacterized phytoene synthase containing pathways and a novel siderophore pathway. Biological assays confirmed that the strain produces molecules with antioxidant and siderophore activities. Furthermore, intracellular extracts showed nematocidal activity towards C. elegans, suggesting that strain KRL4 is a source of anthelmintic compounds.

Prokaryotic Diversity of the Composting Thermophilic Phase: The Case of Ground Coffee Compost.

Waste biomass coming from a local coffee company, which supplied burnt ground coffee after an incorrect roasting process, was employed as a starting material in the composting plant of the Experimental Station of the University of Naples Federico II at Castel Volturno (CE). The direct molecular characterization of compost using 13C-NMR spectra, which was acquired through cross-polarization magic-angle spinning, showed a hydrophobicity index of 2.7% and an alkyl/hydroxyalkyl index of 0.7%. Compost samples that were collected during the early "active thermophilic phase" (when the composting temperature was 63 °C) were analyzed for the prokaryotic community composition and activities. Two complementary approaches, i.e., genomic and predictive metabolic analysis of the 16S rRNA V3-V4 amplicon and culture-dependent analysis, were combined to identify the main microbial factors that characterized the composting process. The whole microbial community was dominated by Firmicutes. The predictive analysis of the metabolic functionality of the community highlighted the potential degradation of peptidoglycan and the ability of metal chelation, with both functions being extremely useful for the revitalization and fertilization of agricultural soils. Finally, three biotechnologically relevant Firmicutes members, i.e., Geobacillus thermodenitrificans subsp. calidus, Aeribacillus pallidus, and Ureibacillus terrenus (strains CAF1, CAF2, and CAF5, respectively) were isolated from the "active thermophilic phase" of the coffee composting. All strains were thermophiles growing at the optimal temperature of 60 °C. Our findings contribute to the current knowledge on thermophilic composting microbiology and valorize burnt ground coffee as waste material with biotechnological potentialities.

Novel Psychrophiles and Exopolymers from Permafrost Thaw Lake Sediments.

Thermokarst lakes are one of the most abundant types of microbial ecosystems in the circumpolar North. These shallow basins are formed by the thawing and collapse of ice-rich permafrost, with subsequent filling by snow and ice melt. Until now, permafrost thaw lakes have received little attention for isolation of microorganisms by culture-based analysis. The discovery of novel psychrophiles and their biomolecules makes these extreme environments suitable sources for the isolation of new strains, including for potential biotechnological applications. In this study, samples of bottom sediments were collected from three permafrost thaw lakes in subarctic Québec, Canada. Their diverse microbial communities were characterized by 16S rRNA gene amplicon analysis, and subsamples were cultured for the isolation of bacterial strains. Phenotypic and genetic characterization of the isolates revealed affinities to the genera Pseudomonas, Paenibacillus, Acinetobacter,Staphylococcus and Sphingomonas. The isolates were then evaluated for their production of extracellular enzymes and exopolymers. Enzymes of potential biotechnological interest included α and ß-glucosidase, α and ß-maltosidase, ß-xylosidase and cellobiohydrolase. One isolate, Pseudomonas extremaustralis strain 2ASCA, also showed the capability to produce, in the loosely bound cell fraction, a levan-type polysaccharide with a yield of 613 mg/L of culture, suggesting its suitability as a candidate for eco-sustainable alternatives to commercial polymers.

Hetero-exopolysaccharide from the extremely halophilic Halomonas smyrnensis K2: production, characterization and functional properties in vitro.

In this study, we firstly reported the production and the structural characterization of a novel hetero-exopolysaccharide namely EPS-K2 from the extremely halophilc Halomonas smyrnensis K2. Results revealed that EPS-K2 was mainly composed of three monosaccharides including mannose (66.69%), glucose (19.54%) and galactose (13.77%). EPS-K2 showed high thermostability with a degradation temperature around 260 °C, which could make it a suitable candidate for application in thermal processes. Moreover, EPS-K2 showed attractive functional properties. In fact, it exhibited potent antioxidant activity in a dose-dependent manner as assessed in analyses of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, iron chelating and DNA protection ability. Furthermore, EPS-K2 showed strong adhesion inhibition activity against Enterococcus faecalis (75.52 ± 3.35%) and Escherichia coli (61.95 ± 2.48%) at 1 g/l concentration, as well as a high biofilm disruption activity especially against E. coli (70.73 ± 2.78%), at 2 g/l concentration. According to its biotechnological properties, EPS-K2 could be exploited as functional ingredient in food, biomedicine, and pharmaceutical industries.

Structural characterization and functional properties of novel exopolysaccharide from the extremely halotolerant Halomonas elongata S6.

Production of extracellular polysaccharides by halophilic Archaea and Bacteria has been widely reported and the members of the genus Halomonas have been identified as the most potential producers. In the present work, a novel exopolysaccharide (EPS-S6) produced by the extremely halotolerant newly isolated Halomonas elongata strain S6, was characterized. According to the HPAE-PAD results, EPS-S6 was mainly composed of glucosamine, mannose, rhamnose and glucose (1:0.9:0.7:0.3). EPS-S6 was highly negatively charged and its molecular weight was about 270 kDa. Studies on its functional properties showed that EPS-S6 had several potential features. It has noticeable antioxidant activities on 2,2-diphenyl-1-picrylhydrazyl (DPPH•) inhibition and DNA protection, good ability to inhibit and to disrupt pathogenic biofilms, excellent flocculation of kaolin suspension and interesting emulsifying properties at acidic, neutral and basic pH. Therefore, EPS-S6 could have potential biotechnological concern in several fields such as in food, cosmetic and environmental industries.

Acinetobacter mesopotamicus sp. nov., Petroleum-degrading Bacterium, Isolated from Petroleum-Contaminated Soil in Diyarbakir, in the Southeast of Turkey.

A new petroleum-degrading bacterium, designated strain GC2T, was isolated from Bozkus 1 petroleum station in Diyarbakir, located in the southeast of Turkey. Cells were Gram-negative staining, aerobic, coccoid-rods, non-motile, non-spore-forming. The bacterium was found to degrade 100% of n-alkanes ranging from C11 to C34 presented in the 1% crude oil after incubation of 7 days. The membrane phospholipids were 1,2 diacylglycero-3-phosphorylethanolamine (PEA), phosphatidylglycerol (PG), dipalmitoyl-sn-glycerol 1- phosphocholine (PC1), 1,2 dipalmitoyl-sn-glycero-3-phosphocholine monohydrate (PC3), cardiolipin also called diphosphatidylglycerol (CL) and l-α- phosphatidic acid, dipalmitoyl (AP); predominant respiratory ubiquinone was Q-8 and C16:0, C18:1ω9c and C16:1 were the major cellular fatty acids. The 16S rRNA sequence analysis revealed that the strain GC2T was a member of genus Acinetobacter and was most closely related to Acinetobacter lwoffii DSM 2403 T (99.79%), Acinetobacter pseudolwoffii ANC 5318 T (98.83%) and Acinetobacter harbinensis HITLi 7 T (98.14%). The rpoB and gyrB gene sequence analysis confirmed that the strain GC2T was a member of genus Acinetobacter and that the closest relative was Acinetobacter lwoffii DSM 2403 T (99.08% and 100% similarity, respectively). DNA-DNA hybridization values between GC2T and its closest relatives ranged from 65.6% (with A. lwoffii) to 5.1% (with A. venetianus). The whole genome sequence of strain GC2T was obtained. The DNA G + C content of this strain was determined to be 42.9 mol %. ANI indexes, in silico estimations of DDH values and wet lab DDH values demonstrated that strain GC2T represents an independent genomospecies. On the basis of phenotypic characteristics, chemotaxonomic, phylogenetic data and DNA-DNA hybridization and whole genome analysis, we propose to assign strain GC2T as a new species of the genus Acinetobacter, for which the name Acinetobacter mesopotamicus sp. nov. is proposed. The type strain of this species is GC2T (DSM 26953 T = JCM 31073 T). The whole genome of strain GC2T has been deposited at DDBJ/ENA/GenBank under the accession JAALFF010000000.

Graphene-Based and Surface-Enhanced Raman Spectroscopy for Monitoring the Physio-Chemical Response of Thermophilic Bacterial Spores to Low Temperatures Exposure.

Monitoring the spore life cycle is one of the main issues in several fields including environmental control, sustainable ecosystems, food security, and healthcare systems. In this framework, the study of the living organism resistance to extreme conditions like those mimicking space environments is particularly interesting. The assessment of the local change of the pH level can be extremely useful for this purpose. An optical physiometer method based on the Raman response of the graphene, which is able to locally sense pH of a fluid on a micrometric scale, has been recently proposed. Due to the presence of π -bonds at the surface, the electronic doping of graphene is determined by the external conditions and can be electrochemically controlled or altered by the contact with an acid or alkaline fluid. The doping level affects the vibrational energies of the graphene that can be monitored by conventional Raman spectroscopy. In addition, Surface-Enhanced Raman Spectroscopy (SERS) can give direct information on the biochemical changes occurring in spore components. In this work, we propose the joint use of Graphene-Based Raman Spectroscopy (GbRS) and SERS for the monitoring of the response of spores to exposure to low temperatures down to 100 K. The spores of the thermophilic bacterium Parageobacillus thermantarcticus isolated from an active volcano of Antarctica (Mt. Melbourne) were investigated. These spores are particularly resistant to several stressing stimuli and able to adapt to extreme conditions like low temperatures, UV irradiation, and γ -rays exposure. The results obtained showed that the joint use of GbRS and SERS represents a valuable tool for monitoring the physio-chemical response of bacterial spores upon exposure to stressing stimuli.

Peculiarities of extracellular polymeric substances produced by Antarctic bacteria and their possible applications.

Extracellular polymeric substances (EPSs) possess diversified ecological role, including the cell adhesion to surfaces and cell protection, and are highly involved in the interactions between the bacterial cells and the bulk environments. Interestingly, EPSs find valuable applications in the industrial field, due to their chemical versatility. In this context, Antarctic bacteria have not been given the attention they deserve as producers of EPS molecules and a very limited insight into their EPS production capabilities and biotechnological potential is available in literature to date. Antarctic EPS-producing bacteria are mainly psychrophiles deriving from the marine environments (generally sea ice and seawater) around the continent, whereas a unique thermophilic bacterium, namely Parageobacillus thermantarcticus strain M1, was isolated from geothermal soil of the crater of Mount Melbourne. This mini-review is aimed at showcasing the current knowledge on EPS-producing Antarctic bacteria and the chemical peculiarities of produced EPSs, highlighting their biotechnological potential and the yet unexplored treasure they represent for biodiscovery.

Vegetable wastes derived polysaccharides as natural eco-friendly plasticizers of sodium alginate.

In this paper, lemon and fennel wastes were recovered and used as secondary-raw polysaccharide sources. These polysaccharides were exploited as natural plasticizers of sodium alginate (A) based films, in order to improve sodium alginate performances, limited by its fragility, extending its potential application in a cost effective and eco-friendly way. Different green processes, such as maceration (MAC), ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE), were carried out for obtaining high yield of lemon and fennel polysaccharides (LP and FP). Actually, HPAE-PAD and TLC analyses evidenced the presence of xyslose, galactose, glucose and rhamnose monomers and galacturonic acid, typical of polysaccharides like pectin and xyloglucan chains. These findings were confirmed by NMR and FTIR spectroscopic analyses. Moreovers, gel filtration chromatography assessed the high molecular weight of recovered polysaccharides, particularly of FP waste fraction. The extracted polysaccharides were used as eco-friendly and cost-effective plasticizers of sodium alginate films (AFP and ALP). DSC analysis evidenced a significant decreasing of glass transition temperature of the polymer, tensile tests showed an enlightened rising of elongation at break and TGA analysis showed a faster degradation kinetics of AFP and ALP films, as expected in a plasticized system.