Summary Biological nitrogen fixation is catalyzed by the molybdenum (Mo), vanadium (V) and iron (Fe)‐only nitrogenase metalloenzymes. Studies with purified enzymes have found that the ‘alternative’ V‐ and Fe‐nitrogenases generally reduce N2 more slowly and produce more byproduct H2 than the Mo‐nitrogenase, leading to an assumption that their usage results in slower growth. Here we show that, in the metabolically versatile photoheterotroph Rhodopseudomonas palustris, the type of carbon substrate influences the relative rates of diazotrophic growth based on different nitrogenase isoforms. The V‐nitrogenase supports growth as fast as the Mo‐nitrogenase on acetate but not on the more oxidized substrate succinate. Our data suggest that this is due to insufficient electron flux to the V‐nitrogenase isoform on succinate compared with acetate. Despite slightly faster growth based on the V‐nitrogenase on acetate, the wild‐type strain uses exclusively the Mo‐nitrogenase on both carbon substrates. Notably, the differences in H2:N2 stoichiometry by alternative nitrogenases (~1.5 for V‐nitrogenase, ~4–7 for Fe‐nitrogenase) and Mo‐nitrogenase (~1) measured here are lower than prior in vitro estimates. These results indicate that the metabolic costs of V‐based nitrogen fixation could be less significant for growth than previously assumed, helping explain why alternative nitrogenase genes persist in diverse diazotroph lineages and are broadly distributed in the environment.

Fluorescence of 2-(N,N-dimethylamino)-6-propionylnaphthalene dyes Badan and Prodan is quenched by tryptophan in Brij 58 micelles as well as in two cytochrome P450 proteins (CYP102, CYP119) with Badan covalently attached to a cysteine residue. Formation of nonemissive complexes between a dye molecule and tryptophan accounts for about 76% of the fluorescence intensity quenching in micelles, the rest is due to diffusive encounters. In the absence of tryptophan, fluorescence of Badan-labeled cytochromes decays with triexponential kinetics characterized by lifetimes of about 100 ps, 700–800 ps, and 3 ns. Site mutation of a histidine residue in the vicinity of the Badan label by tryptophan results in shortening of all three decay lifetimes. The relative amplitude of the fastest component increases at the expense of the two slower ones. The average quenching rate constants are 4.5 × 108 s–1 (CYP102) and 3.7 × 108 s–1 (CYP119), at 288 K. Cyclic voltammetry of Prodan in MeCN shows a reversible reduction peak at −1.85 V vs NHE that becomes chemically irreversible and shifts positively upon addition of water. A quasireversible reduction at −0.88 V was observed in an aqueous buffer (pH 7.3). The excited-state reduction potential of Prodan (and Badan) is estimated to vary from about +0.6 V (vs NHE) in polar aprotic media (MeCN) to approximately +1.6 V in water. Tryptophan quenching of Badan/Prodan fluorescence in CYPs and Brij 58 micelles is exergonic by ≤0.5 V and involves tryptophan oxidation by excited Badan/Prodan, coupled with a fast reaction between the reduced dye and water. Photoreduction is a new quenching mechanism for 2-(N,N-dimethylamino)-6-propionylnaphthalene dyes that are often used as solvatochromic polarity probes, FRET donors and acceptors, as well as reporters of solvation dynamics.

Phaeocystis antarctica is an abundant phytoplankton species in the Southern Ocean, where growth is frequently limited by iron and light. Being able to grow under low iron conditions is essential to the species’ success, but there have been hints that this ability differs among clones. Here, we compare the growth, cell size and chlorophyll a concentrations of four P. antarctica clones cultured under different iron and light conditions. Iron was provided either as unchelated iron (Fe′) or bound to the bacterial siderophore desferrioxamine B, representing, respectively, the most and least bioavailable forms of iron which phytoplankton encounter in the marine environment. The growth rate data demonstrate that the clones vary in their ability to grow using organically bound iron, and that this ability is not related to their ability to grow at low inorganic iron concentrations. These results are consistent at low and high light. Physiologically, only three of the four clones shrink or decrease the concentration of chlorophyll a in response to iron limitation, and only one clone decreases colony formation. Together, our data show that P. antarctica clones 1) respond to the same degree of iron limitation using different acclimation strategies, and 2) vary in their ability to grow under the same external iron and light conditions. This physiological diversity is surprisingly large for isolates of a single phytoplankton species.

Environmental contextVolatile selenium compounds from the oceans may ultimately be an important selenium source for agricultural soils. It has been hypothesised that marine algae are responsible for volatile selenium emissions, but in laboratory experiments, we observed minimal volatile selenium production by two marine algae known to produce large amounts of volatile sulfur. Instead, we found hints that bacterial processes may be important in the production of volatile selenium in the oceans. AbstractVolatile methylated selenium compounds, especially dimethylselenide, are thought to comprise the majority of marine selenium emissions. Despite their potential importance for the global redistribution of this trace element, which is essential for human health, little is known about the algal production of volatile organic selenium compounds. Previous studies have found correlations between dissolved dimethylselenide concentrations, dimethylsulfide concentrations (the sulfur analogue of dimethylselenide) and proxies for algal activity, most notably during a bloom of the coccolithophorid Emiliania huxleyi. In culturing studies, we investigated the ability of three globally important marine algal species, E. huxleyi, Phaeocystis globosa and the diatom Thalassiosira oceanica, to produce dimethylselenide. Despite substantial uptake of selenium and the production of volatile sulfur, E. huxleyi and T. oceanica produced negligible volatile selenium (<2nM). P. globosa produced low amounts of volatile selenium (~8nM), but grew poorly in our laboratory. However, cultures of marine bacteria and mixed bacterial–algal cultures showed that substantial amounts of volatile selenium can be produced in the presence of marine bacteria. In addition, a culture of marine bacteria alone produced ~50nM volatile selenium, far more than axenic cultures of E. huxleyi when exposed to equivalent selenite concentrations. Our results hint that marine algae may be of minor importance in the direct production of volatile selenium in the oceans, and suggest that the production of these compounds in the marine biosphere may instead be controlled by bacterial activity.