Chemical characterization and biological effect of exopolysaccharides synthesized by Antarctic yeasts Cystobasidium ongulense AL101 and Leucosporidium yakuticum AL102 on murine innate immune cells.

The current study aimed to investigate exopolysaccharides (EPSs) produced by two Antarctic yeasts isolated from soil and penguin feathers samples collected on Livingston Island (Antarctica). The strains were identified as belonging to the species Leucosporidium yakuticum (LY) and Cystobasidium ongulense (CO) based on molecular genetic analysis. The EPS production was investigated using submerged cultivation. Different chemical, chromatographic, and spectral analyses were employed to characterize EPSs. LY accumulated 5.5 g/L biomass and 4.0 g/L EPS after 120 h of cultivation, while CO synthesized 2.1 g/L EPS at the end of cultivation, and the biomass amount reached 5.5 g/L. LY-EPS was characterized by a higher total carbohydrate content (80%) and a lower protein content (18%) by comparison with CO-EPS (62%, 30%). The LY-EPS mainly consisted of mannose (90 mol%), whereas CO-EPS had also glucose, galactose, and small amounts of uronic acids (8-5 mol%). Spectral analyses (FT-IR and 1D, 2D NMR) revealed that LY-EPS comprised a typical ß-(1 → 4)-mannan. Branched (hetero)mannan, together with ß/α-glucans constituted the majority of CO-EPS. Unlike LY-EPS, which had a high percentage of high molecular weight populations, CO-EPS displayed a large quantity of lower molecular weight fractions and a higher degree of heterogeneity. LY-EPS (100 ng/mL) elevated significantly interferon gamma (IFN-γ) production in splenic murine macrophages and natural killer (NK) cells. The results indicated that newly identified EPSs might affect IFN-γ signaling and in turn, might enhance anti-infectious responses. The data obtained also revealed the potential of EPSs and yeasts for practical application in biochemical engineering and biotechnology.

Antimicrobial and Antioxidant Potential of Scenedesmus obliquus Microalgae in the Context of Integral Biorefinery Concept.

Small-scale photobioreactors (PBRs) in the inoculum stage were designed with internal (red or green) and external white LED light as an initial step of a larger-scale installation aimed at fulfilling the integral biorefinery concept for maximum utilization of microalgal biomass in a multifunctional laboratory. The specific growth rate of Scenedesmus obliquus (Turpin) Kützing biomass for given cultural conditions was analyzed by using MAPLE software. For the determination of total polyphenols, flavonoids, chlorophyll "a" and "b", carotenoids and lipids, UHPLC-HRMS, ISO-20776/1, ISO-10993-5 and CUPRAC tests were carried out. Under red light growing, a higher content of polyphenols was found, while the green light favoured the flavonoid accumulation in the biomass. Chlorophylls, carotenoids and lipids were in the same order of magnitude in both samples. The dichloromethane extracts obtained from the biomass of each PBR synergistically potentiated at low concentrations (0.01-0.05 mg/mL) the antibacterial activity of penicillin, fluoroquinolones or oregano essential oil against the selected food-borne pathogens (Staphylococcus aureus, Escherichia coli and Salmonella typhimurium) without showing any in vitro cytotoxicity. Both extracts exhibited good cupric ion-reducing antioxidant capacity at concentrations above 0.042-0.08 mg/mL. The UHPLC-HRMS analysis revealed that both extracts contained long chain fatty acids and carotenoids thus explaining their antibacterial and antioxidant potential. The applied engineering approach showed a great potential to modify microalgae metabolism for the synthesis of target compounds by S. obliquus with capacity for the development of health-promoting nutraceuticals for poultry farming.

Genetic transformation of rare Verbascum eriophorum Godr. plants and metabolic alterations revealed by NMR-based metabolomics.

OBJECTIVES: To develop a protocol to transform Verbascum eriophorum and to study the metabolic differences between mother plants and hairy root culture by applying NMR and processing the datasets with chemometric tools. RESULTS: Verbascum eriophorum is a rare species with restricted distribution, which is poorly studied. Agrobacterium rhizogenes-mediated genetic transformation of V. eriophorum and hairy root culture induction are reported for the first time. To determine metabolic alterations, V. eriophorum mother plants and relevant hairy root culture were subjected to comprehensive metabolomic analyses, using NMR (1D and 2D). Metabolomics data, processed using chemometric tools (and principal component analysis in particular) allowed exploration of V. eriophorum metabolome and have enabled identification of verbascoside (by means of 2D-TOCSY NMR) as the most abundant compound in hairy root culture. CONCLUSION: Metabolomics data contribute to the elucidation of metabolic alterations after T-DNA transfer to the host V. eriophorum genome and the development of hairy root culture for sustainable bioproduction of high value verbascoside.

Synthesis and characterization of an exopolysaccharide by antarctic yeast strain Cryptococcus laurentii AL100.

An exopolysaccharide-producing Antarctic yeast strain was selected and identified as Cryptococcus laurentii AL100. The physiological properties of the strain and its ability to utilize and biotransform different carbon sources (pentoses, hexoses, and oligosaccharides) into exopolysaccharide and biomass were investigated. Sucrose was chosen as a suitable and accessible carbon source. The biosynthetic capacity of the strain was studied in its dynamics at different sucrose concentrations (20, 30, 40, and 50 g/L) and temperatures (22 and 24 °C). The maximum biopolymer quantity of 6.4 g/L was obtained at 40 g/L of sucrose, 22 °C temperature and 96-h fermentation duration. The newly synthesized microbial carbohydrate was a heteropolysaccharide having the following monosaccharide composition: arabinose, 61.1%; mannose, 15.0%; glucose, 12.0%; galactose, 5.9%; and rhamnose, 2.8%. It was characterized by polydispersity of the polymer molecule, 60% of it having molecular mass of 4200 Da. The exopolysaccharide demonstrated good emulsifying and stabilizing properties with regard to oil/water emulsions and a pronounced synergistic effect with other hydrocolloids such as xanthan gum, guar gum, and alginate.