The effect of sulfite on the ATP hydrolysis and synthesis activities in chloroplasts and cyanobacterial membrane vesicles can be explained by competition with phosphate.

The effect of sulfite on ATP synthesis and hydrolysis activities is investigated in spinach chloroplasts and in membrane vesicles from the cyanobacterium Synechococcus 6716. Sulfite inhibits phenazine methosulfate-mediated cyclic photophosphorylation both in thiol-modulated chloroplasts and in cyanobacterial membranes with HSO3- (bisulfite) as the active ionic species. The observed inhibition is not due to inhibition of electron transfer or to uncoupling by sulfite. ATP synthesis in cyanobacterial membranes is more sensitive to sulfite when the inorganic phosphate concentration is decreased. This indicates competition between sulfite and phosphate for the same binding site on the ATP synthase. In cyanobacterial membranes sulfite can replace a proton gradient as activator of ATP hydrolysis in the same way as in reduced chloroplasts. By modeling, competition between sulfite and phosphate can fully explain the findings concerning both inhibition and activation.

The effect of sulfite on the ATP hydrolysis and synthesis activity of membrane-bound H(+)-ATP synthase from various species.

The action of sulfite on ATP hydrolysis and synthesis activities is investigated in membrane vesicles prepared from the cyanobacterium Synechococcus 6716, chromatophores from the photosynthetic purple bacterium Rhodospirillum rubrum, membrane vesicles from the related non-photosynthetic bacterium Paracoccus denitrificans, and bovine heart submitochondrial particles. Without any further pretreatment ATP hydrolysis is stimulated by sulfite in all four membrane preparations. Typically ATP synthesis in the cyanobacterial membrane vesicles is inhibited by sulfite, whereas ATP synthesis in chromatophores and the submitochondrial particles is not. These differences in sensitivity of ATP synthesis to sulfite, however, correspond well with the distribution of (photosynthetic) sulfur oxidizing pathways in the remaining three organisms/organelles compared in this study.

On the activation mechanism of the H(+)-ATP synthase and unusual thermodynamic properties in the alkalophilic cyanobacterium Spirulina platensis.

The activation requirements and thermodynamic characteristics of ATP synthase from the alkalophilic cyanobacterium Spirulina platensis were studied in coupled membrane vesicles. Activation by methanol increased the Vmax, while the Km for MgATP was unaffected (0.7 mM). We propose that in Sp. platensis, as in chloroplasts, the activating effect of methanol is based on perturbation of the gamma-epsilon subunit interaction. Light-driven ATP synthesis by membrane vesicles of Sp. platensis was stimulated by dithiothreitol. The characteristics of the activation of the ATP synthase by the proton electrochemical potential difference (delta mu H+) were analyzed on the basis of the uncoupled rates of ATP hydrolysis as a function of a previously applied proton gradient. Two values of delta mu H+, at which 50% of the enzyme is active, were found; 13-14 kJ.mol-1 for untreated membrane vesicles, and 4-8 kJ.mol-1 for light-treated and dithiothreitol-treated membrane vesicles. These values are lower than the corresponding values for the oxidized and reduced forms, respectively, of the chloroplast enzyme. Although no bulk proton gradient could be observed, membrane vesicles of Sp. platensis were able to maintain an equilibrium phosphate potential (delta Gp) of 40-43.5 kJ.mol-1, comparable to values found for Synechococcus 6716 and Anabaena 7120 membrane vesicles. Acid/base-transition experiments showed that the thermodynamic threshold, delta mu H+, for ATP synthesis, catalyzed by light-treated and dithiothreitol-treated Spirulina membrane vesicles, was less than 5 kJ.mol-1. The activation characteristics and the low thermodynamic threshold allow ATP synthesis to occur at low delta mu H+ values. The findings are discussed, both with respect to differences and similarities with the enzymes from chloroplasts and other cyanobacteria, and with respect to the alkalophilic properties of Sp. platensis.

Growth yield, maintenance requirements, and lipid formation in the oleaginous yeast Apiotrichum curvatum.

For guiding and improving the efficiency of the production of lipid, complete insight into the flow of carbon and energy during growth and product formation is necessary. Therefore, data have been collected to determine various important growth parameters for the oleaginous yeast Apiotrichum curvatum. Chemostat experiments at specific growth rates, ranging from 0.05 to 0.15 h(-1) and recycling experiments with 100% biomass retention, with growth rates decreasing from 0.10 to 0.004 h(-1), demonstrated that maintenance requirements of A. curvatum are very low, compared to maintenance requirements described for other yeasts as Saccharomyces cerevisiae and Candida parapsilosis.It also appeared that growth and lipid production are proportional to substrate consumption when specific growth rates are higher than approximately 0.02 h(-1), but that lipid production stops at growth rates below this value. The practical consequences of these data are that fed batch cultures, which are often applied in fermentation industry, can only be useful with lipid producing yeasts when the growth rate in the process is carefully monitored to ensure specific growth rates higher than 0.02 h(-1). Dilution of the culture, partial recycling and/or a continuously increasing nutrient feed are solutions for the expected problems at low growth rates.