Acid base regulation during ammonium assimilation in Hydrodictyon africanum

The acid-base balance during ammonium (used to mean NH 4+ and/or NH3) assimilation in Hydrodictyon africanum has been measured on cells growing with about 1 mol m−3 ammonium at an external pH of about 6.5. Measurements made included (1) ash alkalinity (corrected for intracellular ammonium) which yields net organic negative charge, (2) the accumulation of organic N in the cells and (3) the change in extracellular H+ (from the pH change and the buffer capacity). These measurements showed that some 0.25 excess organic negative charge (half in the cell wall, half inside the plasmalemma) accumulates per organic N synthesized, while some 1.25H+ accumulate in the medium per organic N synthesized. Granted a permeability (PNH3) of some 10−3 cm s−1, and a finite [NH3] in the cytoplasm of these N-assimilating cells it is likely that most of the ammonium entering these growing cells is as NH 4+. This means that most of the H + appearing in the medium must have originated from inside the cell and have been subjected to active efflux at the plasmalemma: H+ accumulates in the medium equivalent to any NH3 entry by requilibration from exogenous NH 4+. The cell composition (net organic negative charge, organic N content) is very similar in these ammonium-grown cells to that of NO3+grown cells, suggesting that there is no action of a ‘biochemical pH stat’ during longterm assimilation of NO3+in H. africanum. Short-term experiments were carried out at an external pH of 7.2 in which ammonium at various concentrations were supplied to NO3+-grown cells. There was in all cases a rapid influx followed by a slower uptake; at least at the lower concentrations (less than 100 μmol dm−3) the net influx was all attributable to NH4+influx via a uniporter, probably partly short-circuited by a passive NH3 efflux due to intrinsic membrane permeability to NH3. The net ammonium influx was in all cases associated with H+ accumulation in the medium. (1.3-1.7 H + per ammonium taken up); as in the growth experiments, most of the ammonium taken up was assimilated. Determinations of cytoplasmic pH showed either no effect on, or a slight decrease in, pH during ammonium assimilation; the changes that occurred were in the direction expected for actuating a ‘pH-regulating’ change in H+ fluxes.