Rapid screening of phenolic compounds in extracts of photosynthetic organisms separated using a C18 monolithic column based HPLC-UV method.

The sourcing of novel bioactive compounds from bioresources has become a sustainability priority for several industrial sectors. Reliable methods are hence needed for the screening and identification of desired molecules in complex extracts. As such, phenolic compounds have attracted increasing interests due to their antioxidant potential. Here, a HPLC-UV based method was developed for the determination of phenolic compounds using a monolithic column. This allowed the separation of 23 phenolic compounds within 11 min. The method was ground proofed in terms of chromatographic parameters and applied to identify polyphenols in extracts of blueberries, basil and marine microalgae. The dominant polyphenols in blueberries and basil were chlorogenic acid (26.0 mg. kg -1) and kaempferol (61.4 mg. kg -1), respectively, while the microalgae used only contained trace amounts of polyphenols. This study demonstrates the value of our new HPLC method to rapidly screen extracts and identify polyphenols from biological matrices.

Microbial Poly-γ-Glutamic Acid (γ-PGA) as an Effective Tooth Enamel Protectant.

Poly-γ-glutamic acid (γ-PGA) is a bio-derived water-soluble, edible, non-immunogenic nylon-like polymer with the biochemical characteristics of a polypeptide. This Bacillus-derived material has great potential for a wide range of applications, from bioremediation to tunable drug delivery systems. In the context of oral care, γ-PGA holds great promise in enamel demineralisation prevention. The salivary protein statherin has previously been shown to protect tooth enamel from acid dissolution and act as a reservoir for free calcium ions within oral cavities. Its superb enamel-binding capacity is attributed to the L-glutamic acid residues of this 5380 Da protein. In this study, γ-PGA was successfully synthesised from Bacillus subtilis natto cultivated on supplemented algae media and standard commercial media. The polymers obtained were tested for their potential to inhibit demineralisation of hydroxyapatite (HAp) when exposed to caries simulating acidic conditions. Formulations presenting 0.1, 0.25, 0.5, 0.75, 1, 2, 3 and 4% (w/v) γ-PGA concentration were assessed to determine the optimal conditions. Our data suggests that both the concentration and the molar mass of the γ-PGA were significant in enamel protection (p = 0.028 and p < 0.01 respectively). Ion Selective Electrode, combined with Fourier Transform Infra-Red studies, were employed to quantify enamel protection capacity of γ-PGA. All concentrations tested showed an inhibitory effect on the dissolution rate of calcium ions from hydroxyapatite, with 1% (wt) and 2% (wt) concentrations being the most effective. The impact of the average molar mass (M) on enamel dissolution was also investigated by employing commercial 66 kDa, 166 kDa, 440 kDa and 520 kDa γ-PGA fractions. All γ-PGA solutions adhered to the surface of HAp with evidence that this remained after 60 min of continuous acidic challenge. Inductively Coupled Plasma analysis showed a significant abundance of calcium ions associated with γ-PGA, which suggests that this material could also act as a responsive calcium delivery system. We have concluded that all γ-PGA samples tested (commercial and algae derived) display enamel protection capacity regardless of their concentration or average molar mass. However, we believe that γ-PGA D/L ratios might affect the binding more than its molar mass.

Antioxidant Bioprospecting in Microalgae: Characterisation of the Potential of Two Marine Heterokonts from Irish Waters.

Microalgae constitute a heterogeneous and diverse range of organisms capable of accumulating bioactive metabolites, making them promising feedstock for applications in the nutraceutical, functional food, animal feed, biofertilisation or biofuel sectors. There has been renewed interest in recent times in natural sources of antioxidants, particularly as health products and preserving agents. Microalgae strains isolated from aquatic habitats in Ireland were successfully brought into culture. The 91 strains were grown phototrophically in nutrient-enriched media to generate biomass, which was harvested and assessed for antioxidant potential. Extracts were screened for antioxidant activity using a modified volumetric Trolox-ABTS assay and the Folin-Ciocalteu method. Two heterokont marine strains of interest were further studied to ascertain variations in antioxidant capacity across different stages of batch culture growth. The antioxidant activity of extracts of bacillariophyte cf. Stauroneis sp. LACW24 and ocrophyte cf. Phaeothamnion sp. LACW34 increased during growth with a maximum being observed during the late stationary or early death phase (2.5- to 8-fold increases between days 20 and 27). Strains LACW24 and LACW34 contained 5.9 and 3.0 mg g-1 (DW) of the xanthophyll fucoxanthin, respectively. Extracts of strains also showed no cytotoxicity towards mouse cell lines. These results highlight the potential of these strains for biomass valorisation and cultivation upscaling and to be further considered as part of ongoing bioprospecting efforts towards identifying novel species to join the relatively narrow range of commercially exploited marine microalgae species.

Influence of spectral intensity and quality of LED lighting on photoacclimation, carbon allocation and high-value pigments in microalgae.

Tailoring spectral quality during microalgal cultivation can provide a means to increase productivity and enhance biomass composition for downstream biorefinery. Five microalgae strains from three distinct lineages were cultivated under varying spectral intensities and qualities to establish their effects on pigments and carbon allocation. Light intensity significantly impacted pigment yields and carbon allocation in all strains, while the effects of spectral quality were mostly species-specific. High light conditions induced chlorophyll photoacclimation and resulted in an increase in xanthophyll cycle pigments in three of the five strains. High-intensity blue LEDs increased zeaxanthin tenfold in Rhodella sp. APOT_15 relative to medium or low light conditions. White light however was optimal for phycobiliprotein content (11.2 mg mL-1) for all tested light intensities in this strain. The highest xanthophyll pigment yields for the Chlorophyceae were associated with medium-intensity blue and green lights for Brachiomonas submarina APSW_11 (5.6 mg g-1 lutein and 2.0 mg g-1 zeaxanthin) and Kirchneriella aperta DMGFW_21 (1.5 mg g-1 lutein and 1 mg g-1 zeaxanthin), respectively. The highest fucoxanthin content in both Heterokontophyceae strains (2.0 mg g-1) was associated with medium and high white light for Stauroneis sp. LACW_24 and Phaeothamnion sp. LACW_34, respectively. This research provides insights into the application of LEDs to influence microalgal physiology, highlighting the roles of low light on lipid metabolism in Rhodella sp. APOT_15, of blue and green lights for carotenogenesis in Chlorophyceae and red light-induced photoacclimation in diatoms.