Cu transport and complexation by the marine diatom Phaeodactylum tricornutum: Implications for trace metal complexation kinetics in the surface ocean.

Elucidating whether dissolved Cu uptake is kinetically or thermodynamically controlled, and the effects of speciation on Cu transport by phytoplankton will allow better modeling of the fate and impact of dissolved Cu in the ocean. To address these questions, we performed Cu physiological and physicochemical experiments using the model diatom, Phaeodactylum tricornutum, grown in natural North Atlantic seawater (0.44 nM Cu). Using competitive ligand equilibration-cathodic stripping voltammetry (CLE-CSV), we measured two organic ligand types released by P. tricornutum to bind Cu (L1 and L2) at concentrations of ~0.35 nM L1 and 1.3 nM L2. We also established the presence of two putative Cu-binding sites at the cell surface of P. tricornutum (S1 and S2) with log K differing by ~5 orders of magnitude (i.e., 12.9 vs. 8.1) and cell surface densities by 9-fold. Only the high-affinity binding sites, S1, exhibit reductase activity. Using voltammetric kinetic measurements and a theoretical kinetic model, we calculated the forward and dissociation rate constants of L1 and S1. Complementary 67Cu uptake experiments identified a high- and a low-affinity Cu uptake system in P. tricornutum, with half-saturation constant (Km) of 154 nM and 2.63 µM dissolved Cu, respectively. In the P. tricornutum genome, we identified a putative high-affinity Cu transporter (PtCTR49224) and a putative ZIP-like, low-affinity Cu transporter (PtZIP49400). PtCTR49224 has high homology to Homo sapiens hCTR1, which depending on the accessibility to extracellular reducing agents, the hCTR1 itself is involved in the reduction of Cu2+ to Cu+ before internalization. We combined these physiological and physicochemical data to calculate the rate constants for the internalization of Cu, and established that while the high-affinity Cu uptake system (S1) is borderline between a kinetically or thermodynamically controlled system, the low-affinity Cu transporters, S2, is thermodynamically-controlled. We revised the inverse relationship between the concentrations of inorganic complexes of essential metals (i.e., Ni, Fe, Co, Zn, Cd, Mn and Cu) in the mixed layer and the formation rate constant of metal transporters in phytoplankton, highlighting the link between the chemical properties of phytoplankton metal transporters and the availability and speciation of trace metals in the surface ocean.

Exploring Pavlova pinguis chemical diversity: a potentially novel source of high value compounds.

To uncover the potential of Pavlova pinguis J.C. Green as a natural source of value added compounds, its lipophilic extracts were studied before and after alkaline hydrolysis using gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis of the lipophilic extracts showed a wide chemical diversity including 72 compounds distributed by fatty acids (29), sterols (14), fatty alcohols (13) and other lipophilic compounds (16). Fatty acids represented the main class of identified compounds presenting myristic, palmitic, palmitoleic and eicosapentaenoic acids as its main components. Through the ∑ω6/∑ω3 ratio (0.25) and sterol composition it was possible to observe that P. pinguis is a valuable source of ω3 fatty acids and stigmasterol (up to 43% of total sterols). After alkaline hydrolysis, fatty acids and fatty alcohols content increased by 32 and 14% respectively, in contrast to, monoglycerides which decreased by 84%. The long chain alcohols content enables the exploitation of this microalga as a source of these bioactive compounds. Smaller amounts of sugars and other compounds were also detected. The present study is a valuable reference to the metabolite characterization of P. pinguis and shows the potential of this microalga for nutraceutical and pharmaceutical industries.