RESUMEN
Microalgae are the richest source of natural carotenoids-accessory photosynthetic pigments used as natural antioxidants, safe colorants, and nutraceuticals. Microalga Bracteacoccus aggregatus IPPAS C-2045 responds to stresses, including high light, with carotenogenesis-gross accumulation of secondary carotenoids (the carotenoids structurally and energetically uncoupled from photosynthesis). Precise mechanisms of cytoplasmic transport and subcellular distribution of the secondary carotenoids under stress are still unknown. Using multimodal imaging combining micro-Raman imaging (MRI), fluorescent lifetime (τ) imaging (FLIM), and transmission electron microscopy (TEM), we monitored ultrastructural and biochemical rearrangements of B. aggregatus cells during the stress-induced carotenogenesis. MRI revealed a decline in the diversity of molecular surrounding of the carotenoids in the cells compatible with the relocation of the bulk of the carotenoids in the cell from functionally and structurally heterogeneous photosynthetic apparatus to the more homogenous lipid matrix of the oleosomes. Two-photon FLIM highlighted the pigment transformation in the cell during the stress-induced carotenogenesis. The structures co-localized with the carotenoids with shorter τ (mainly chloroplast) shrunk, whereas the structures harboring secondary carotenoids with longer τ (mainly oleosomes) expanded. These changes were in line with the ultrastructural data (TEM). Fluorescence of B. aggregatus carotenoids, either in situ or in acetone extracts, possessed a surprisingly long lifetime. We hypothesize that the extension of τ of the carotenoids is due to their aggregation and/or association with lipids and proteins. The propagation of the carotenoids with prolonged τ is considered to be a manifestation of the secondary carotenogenesis suitable for its non-invasive monitoring with multimodal imaging.
Asunto(s)
Carotenoides , Microalgas , Estrés Fisiológico , Carotenoides/metabolismo , Microalgas/metabolismoRESUMEN
Terrestrial algae are a group of photosynthetic organisms that can survive in extreme conditions. pH is one of the most important factors influencing the distribution of algae in both aquatic and terrestrial ecosystems. The impact of different pH levels on the cell volume and other morphological characteristics of authentic and reference strains of Chlorella vulgaris, Bracteacoccus minor, Pseudoccomyxa simplex, Chlorococcum infusionum, and Vischeria magna were studied. Chlorella vulgaris, Pseudoccomyxa simplex, and Vischeria magna were the most resistant species, retaining their morphology in the range of pH 4-11.5 and pH 3.5-11, respectively. The change in pH towards acidic and alkaline levels caused an increase in the volume of Pseudoccomixa simplex and Vischeria magna cells, according to a polynomial regression model. The volume of Chlorella vulgaris cells increased from a low to high pH according to a linear regression model. Changes in pH levels did not have a significant impact on the volume of Bracteacoccus minor and Chlorococcum infusionum cells. Low and high levels of pH caused an increase in oil-containing substances in Vischeria magna and Bracteacoccus minor cells. Our study revealed a high resistance of the studied species to extreme pH levels, which allows for us to recommend these strains for broader use in biotechnology and conservation studies of natural populations.
RESUMEN
Microalgae have the highest capability to fix the atmospheric carbon and wastewater-derived nutrients to produce high-value bioproducts including lipids and carotenoids. However, their lower titers and single-product-oriented biomass processing have made the overall process expensive. Hence, increased metabolite titer and processing of the biomass for more than one product are required to ensure the commercial robustness of the algal biorefinery. In this study, a newly isolated algal strain was identified as Bracteacoccus pseudominor BERC09 through phylogenetic analysis based on the 18S rRNA gene sequence. Basic characterization of the strain revealed its promising potential to produce carotenoids and lipids. The lipids and carotenoid biosynthesis pathways of BERC09 were further triggered by manipulating the abiotic factors including nitrogen sources (NaNO3, KNO3, NH4Cl, Urea), nitrogen concentrations (0.06-0.36 gL-1), light intensity (150 µmolm-2s-1 to 300 µmolm-2s-1), and light quality (white and blue). Resultantly, 300 µmolm-2s-1 of blue light yielded 0.768 gL-1 of biomass, 8.4 mgg-1 of carotenoids, and 390 mgg-1 of lipids, and supplementation of 0.36 gL-1 of KNO3 further improved metabolism and yielded 0.814 gL-1 of biomass, 11.86 mgg-1 of carotenoids, and 424 mgg-1 of lipids. Overall, the optimal combination of light and nitrogen concurrently improved biomass, carotenoids, and lipids by 3.5-fold, 6-fold, and 4-fold than control, respectively. Besides, the excellent glycoproteins-based self-flocculation ability of the strain rendered an easier harvesting via gravity sedimentation. Hence, this biomass can be processed in a cascading fashion to use this strain as a candidate for a multiproduct biorefinery to achieve commercial robustness and environmental sustainability.
Asunto(s)
Chlorophyceae , Microalgas , Biomasa , Carotenoides/metabolismo , Chlorophyceae/metabolismo , Lípidos , Microalgas/metabolismo , Nitrógeno/metabolismo , FilogeniaRESUMEN
UV-A is the main ultraviolet component of natural (solar) radiation. Despite it, its effect on phototrophs is studied less than UV-B. Effects of UV-A on photosynthetic apparatus of three carotenoid-producing microalgae were elucidated. Photosynthetic activity was studied using chlorophyll fluorescence analysis. Cell extracts were evaluated by absorbance spectroscopy. On the one hand, there were some common features of three strains. In all cases the changes involved PSII primary photochemistry and antennae size. All strains accumulated UV-absorbing polar compounds. On the other hand, some responses were different. Upregulation of non-photochemical quenching was observed only in B. aggregatus BM5/15, whereas in other cases its level was low. H. rubicundus BM7/13 and Deasonia sp. NAMSU 934/2 accumulated secondary carotenoids, whereas B. aggregatus BM5/15 accumulated primary ones. Microscopic features of the cultures were also different. H. rubicundus BM7/13 and Deasonia sp. NAMSU 934/2 were represented mostly by solitaire cells or small cell clusters, lacking their green color; the cells of B. aggregatus BM5/15 formed aggregates from green cells. Cell aggregation could be considered as an additional UV-protecting mechanism. Finally, the strains differed by their viability. B. aggregatus BM5/15 was most resistant to UV-A, whereas massive cell death was observed in two other cultures.
RESUMEN
Carotenoids astaxanthin and ß-carotene are widely used natural antioxidants. They are key components of functional food, cosmetics, drugs and animal feeding. They hold leader positions on the world carotenoid market. In current work, we characterize the new strain of the green microalga Bracteacoccus aggregatus BM5/15 and propose the method of its culturing in a bubble-column photobioreactor for simultaneous production of astaxanthin and ß-carotene. Culture was monitored by light microscopy and pigment kinetics. Fatty acid profile was evaluated by tandem gas-chromatography-mass spectrometry. Pigments were obtained by the classical two-stage scheme of autotrophic cultivation. At the first, vegetative, stage biomass accumulation occurred. Maximum specific growth rate and culture productivity at this stage were 100-200 mgâL-1âday-1, and 0.33 day-1, respectively. At the second, inductive, stage carotenoid synthesis was promoted. Maximal carotenoid fraction in the biomass was 2.2-2.4%. Based on chromatography data, astaxanthin and ß-carotene constituted 48 and 13% of total carotenoid mass, respectively. Possible pathways of astaxanthin synthesis are proposed based on carotenoid composition. Collectively, a new strain B. aggregatus BM5/15 is a potential biotechnological source of two natural antioxidants, astaxanthin and ß-carotene. The results give the rise for further works on optimization of B. aggregatus cultivation on an industrial scale.
RESUMEN
Carotenogenic microalgae are unicellular photosynthetic organisms with the ability to accumulate carotenoids. Carotenoid accumulation is a protective reaction against environmental stress factors, such as bright light and extreme temperatures. It makes the survival of these microorganisms under harsh environmental conditions possible. The diversity of carotenogenic microalgae has been described in detail for Central Europe and North America, as well as for tropical and subtropical latitudes with relatively favorable environments. However, data about these microorganisms in polar and subpolar latitudes are scarce and restricted to few reports. We isolated several strains of carotenogenic microalgae from the coastal zone of the White Sea, where they were abundant. The obtained microalgae related to four species of Chlorophytes: Haematococcus lacustris, H. rubicundus, Coelastrella aeroterrestrica and Bracteacoccus aggregatus. The last three species have been reported for polar latitudes for the first time. Most likely, carotenogenic algae in the White Sea coast are abundant due to their high physiological and metabolic plasticity, which is essential for surviving under adverse conditions of the northern regions. Pigment composition of the strains is provided. Their predominant carotenoids were astaxanthin and ß-carotene. Further, the obtained strains may be considered as potential producers of natural pigments for biotechnology.