Isolation of Mycosporine-like Amino Acids from Red Macroalgae and a Marine Lichen by High-Performance Countercurrent Chromatography: A Strategy to Obtain Biological UV-Filters.

Marine organisms have gained considerable biotechnological interest in recent years due to their wide variety of bioactive compounds with potential applications. Mycosporine-like amino acids (MAAs) are UV-absorbing secondary metabolites with antioxidant and photoprotective capacity, mainly found in organisms living under stress conditions (e.g., cyanobacteria, red algae, or lichens). In this work, five MAAs were isolated from two red macroalgae (Pyropia columbina and Gelidium corneum) and one marine lichen (Lichina pygmaea) by high-performance countercurrent chromatography (HPCCC). The selected biphasic solvent system consisted of ethanol, acetonitrile, saturated ammonium sulphate solution, and water (1:1:0.5:1; v:v:v:v). The HPCCC process for P. columbina and G. corneum consisted of eight separation cycles (1 g and 200 mg of extract per cycle, respectively), whereas three cycles were performed for of L. pygmaea (1.2 g extract per cycle). The separation process resulted in fractions enriched with palythine (2.3 mg), asterina-330 (3.3 mg), shinorine (14.8 mg), porphyra-334 (203.5 mg) and mycosporine-serinol (46.6 mg), which were subsequently desalted by using precipitation with methanol and permeation on a Sephadex G-10 column. Target molecules were identified by HPLC, MS, and NMR.

Biomass, photosynthetic activity, and biomolecule composition in Chlorella fusca (Chlorophyta) cultured in a raceway pond operated under greenhouse conditions.

Raceways are widely used as microalgae culture systems due to their low cost, but they are not the best option for biomass yield. Understanding in situ photosynthetic performance can be a first step to increase their biomass productivity. This study aimed at comparing the real time photosynthetic activity in a greenhouse raceway culture (250 L) with discrete measurements under laboratory conditions. We evaluated the photophysiology and biochemical composition of Chlorella fusca culture up to 120 h. In situ photosynthetic activity was continuously monitored and compared to discrete ex situ measurements; biochemical compounds were measured daily. The results showed a final biomass density of 0.45 g L-1 (5 days - 120 h) and an increase of the electron transport rate (ETR) up to 48 h but decreased thereafter. When the relative ETR was estimated considering the absorption coefficient (a) positive correlations of this parameter with photosynthetic capacity, cell density, biomass, biocompounds and antioxidant activity were obtained, whereas no correlation was detected without considering a. In situ photosynthesis monitoring showed higher absolute maximal ETR (10 - 160 µmol m-3s-1) than discrete ex situ measurements. We demonstrated the importance of considering the light absorption coefficient for expressing photosynthetic capacity and showed that C. fusca can produce, in the short-term, bioactive compounds that are correlated to photosynthetic conditions.

Holistic Photoprotection, Broad Spectrum (UVA-UVB), and Biological Effective Protection Factors (BEPFs) from Baccharis antioquensis Hydrolysates Polyphenols.

Overexposure to solar radiation has become an increasingly worrying problem due to the damage to the skin caused by ultraviolet radiation (UVR). In previous studies, the potential of an extract enriched with glycosylated flavonoids from the endemic Colombian high-mountain plant Baccharis antioquensis as a photoprotector and antioxidant was demonstrated. Therefore, in this work we sought to develop a dermocosmetic formulation with broad-spectrum photoprotection from the hydrolysates and purified polyphenols obtained from this species. Hence, the extraction of its polyphenols with different solvents was evaluated, followed by hydrolysis and purification, in addition to the characterization of its main compounds by HPLC-DAD and HPLC-MS, and evaluation of its photoprotective capacity through the measurement of the Sun Protection Factor (SPF), UVA Protection Factor (UVAPF), other Biological Effective Protection Factors (BEPFs), and its safety through the cytotoxicity. In the dry methanolic extract (DME) and purified methanolic extract (PME), flavonoids such as quercetin and kaempferol were found, which demonstrated antiradical capacity, as well as UVA-UVB photoprotection and prevention of harmful biological effects, such as elastosis, photoaging, immunosuppression, DNA damage, among others, which demonstrates the potential of the ingredients in this type of extract to be applied in photoprotection dermocosmetics.

Photosynthetic performance of Chlamydopodium (Chlorophyta) cultures grown in outdoor bioreactors.

The microalga Chlamydopodium fusiforme MACC-430 was cultured in two types of outdoor pilot cultivation units-a thin-layer cascade (TLC) and a raceway pond (RWP) placed in a greenhouse. This case study aimed to test their potential suitability for cultivation scale-up to produce biomass for agriculture purposes (e.g., as biofertilizer or biostimulant). The culture response to the alteration of environmental conditions was evaluated in "exemplary" situations of good and bad weather conditions using several photosynthesis measuring techniques, namely oxygen production, and chlorophyll (Chl) fluorescence. Validation of their suitability for online monitoring in large-scale plants has been one of the objectives of the trials. Both techniques were found fast and robust reliable to monitor microalgae activity in large-scale cultivation units. In both bioreactors, Chlamydopodium cultures grew well in the semi-continuous regime using daily dilution (0.20-0.25 day-1). The biomass productivity calculated per volume was significantly (about 5 times) higher in the RWPs compared to the TLCs. The measured photosynthesis variables showed that the build-up of dissolved oxygen concentration in the TLC was higher, up to 125-150% of saturation (%sat) as compared to the RWP (102-104%sat). As only ambient CO2 was available, its shortage was indicated by a pH increase due to photosynthetic activity in the thin-layer bioreactor at higher irradiance intensities. In this setup, the RWP was considered more suitable for scale-up due to higher areal productivity, lower construction and maintenance costs, the smaller land area required to maintain large culture volumes, as well as lower carbon depletion and dissolved oxygen build-up. KEY POINTS: • Chlamydopodium was grown in both raceways and thin-layer cascades in pilot-scale. • Various photosynthesis techniques were validated for growth monitoring. • In general, raceway ponds were evaluated as more suitable for cultivation scale-up.

UV responses of Lolium perenne raised along a latitudinal gradient across Europe: a filtration study.

Lolium perenne (cv. AberDart) was grown at 14 locations along a latitudinal gradient across Europe (37-68°N) to study the impact of ultraviolet radiation (UV) and climate on aboveground growth and foliar UV-B absorbing compounds. At each location, plants were grown outdoors for 5 weeks in a replicated UV-B filtration experiment consisting of open, UV-B transparent (cellulose diacetate) and UV-B opaque (polyester) environments. Fourier transform-infrared spectroscopy was used to compare plant metabolite profiles in relation to treatment and location. UV radiation and climatic parameters were determined for each location from online sources and the data were assessed using a combination of anova and multiple regression analyses. Most of the variation in growth between the locations was attributable to the combination of climatic parameters, with minimum temperature identified as an important growth constraint. However, no single environmental parameter could consistently account for the variability in plant growth. Concentrations of foliar UV-B absorbing compounds showed a positive trend with solar UV across the latitudinal gradient; however, this relationship was not consistent in all treatments. The most striking experimental outcome from this study was the effect of presence or absence of filtration frames on UV-absorbing compounds. Overall, the study demonstrates the value of an European approach in studying the impacts of natural UV across a large latitudinal gradient. We have shown the feasibility of coordinated UV filtration at multiple sites but have also highlighted the need for open controls and careful interpretation of plant responses.