Do the higher oxidation states of the photosynthetic O2-evolving system contain bound H2O?

A modified mass spectrometer was used to determine whether the higher oxidation states of the photosynthetic O2-evolving system contain substrate water that is not freely exchangeable with the external medium. Our data indicated that the higher oxidation states contain no appreciable bound, non-exchangeable H2O. This suggests that H2O oxidation takes place via a rapid, concerted, all-or-none mechanism rather than by a mechanism involving stable, partially oxidized, H2O-derived intermediates. These findings set definite constraints on possible mechanisms of O2 evolution.

Algal culture studies related to a closed ecological life support system.

Long-term cultures of Scenedesmus obliquus were maintained in an annular air-lift column operated as a turbidostat. We observed a linear relationship between the dry weight of the cultured cells, their cell number, and their chlorophyll content over a broad range of cell density at constant illumination. Thus, the cells did not appear to be adapting to differences in growth rate or light intensity during these experiments. Productivity vs dry wt rose linearly until the cell density reached a level at which light became limiting; at this point approximately 89% of the photosynthetically active radiation (PAR) was being absorbed. The maximum dilution rate of the system corresponded to a doubling time of 13.8 hr, about half the maximum growth rate generally observed at this temperature. Productivity at the maximum was approximately 80% of the maximum theoretical productivity. The rather low incident intensities (approximately 10% of full sunlight) were a main contributing factor to the high light utilization efficiencies obtained in this system, since the cells were never driven into light saturation. In many respects, algae would be ideal plant components for a biologically-based closed life support system, since they are eminently suited to the closely coupled functions of food production and atmosphere regeneration. In this communication, we report some findings on the (steady-state) continuous culture of Scenedesmus obliquus, a physiologically well-characterized green alga with good growth characteristics.

Light-driven Uptake of Oxygen, Carbon Dioxide, and Bicarbonate by the Green Alga Scenedesmus.

Mass spectrometric techniques were used to study several aspects of the competition between O(2) and species of inorganic carbon for photosynthetically generated reducing power in the green alga, Scenedesmus.In contrast to wild type, no appreciable light-driven O(2) uptake was observed in a mutant lacking photosystem I. It is concluded that the carbon cycle-independent reduction of O(2) occurs at the expense of photosystem I-generated reducing equivalents.The commonly observed differences between CO(2)-grown and air-grown Scenedesmus with respect to CO(2) uptake and glycolate formation cannot be ascribed to differences in their capacity for light-driven O(2) uptake. There were no intrinsic differences found in O(2) uptake capacity between the two physiological types under conditions in which CO(2) was saturating or CO(2) uptake was inhibited. It was only under CO(2)-limited conditions that pronounced differences between the two physiological types were observed. This fact suggests that differences in O(2) metabolism and sensitivity between the two types really reflect differences in their capacity to assimilate inorganic carbon; in this respect they are analogous to C(3) and C(4) plants.The hypothesis that air-grown Scenedesmus can assimilate HCO(3) (-) by directly monitoring the time course of dissolved CO(2), O(2) uptake, and O(2) evolution in illuminated algal suspensions at alkaline pH was tested. Inasmuch as the measuring technique employed was fast compared to the nonenzymic equilibration of the inorganic carbon species, it was possible to determine the degree to which the CO(2) concentration deviated from equilibrium (with the other inorganic carbon species) during the course of illumination. The observed kinetics in air-grown and CO(2)-grown algae in the presence and absence of carbonic anhydrase, and a comparison of these kinetics with theoretical (computer-generated) time courses, support the idea that air-adapted algae are able to assimilate HCO(3) (-) actively at a high rate. The data suggest that these algae preferentially assimilate CO(2) and supply the balance of their needs by taking up HCO(3) (-). Since (unlike C(4) plants) these algae have no special CO(2) pump, and thus have a relatively low affinity for CO(2), HCO(3) (-) assimilation is the major carbon uptake process at alkaline pH even when the total CO(2) is present in millimolar concentrations.

Kinetics and Apparent K(m) of Oxygen Cycle under Conditions of Limiting Carbon Dioxide Fixation.

A mass spectrometer with a membrane inlet was used to monitor light-driven O(2) evolution, O(2) uptake, and CO(2) uptake in suspensions of algae (Scenedesmus obliquus). We observed the following. (a) The rate of O(2) uptake, which, in the presence of iodoacetamide, replaces the uptake of CO(2), showed a distinct plateau (V(max)) beyond approximately 30% O(2) and was half-maximal at approximately 8% O(2). We concluded that this light-driven O(2) uptake process, which does not involve carbon compounds, is saturated at lower O(2) concentrations than are photorespiration and glycolate formation. (b) In the absence of inhibitor, O(2) evolution was relatively unaffected by the presence or absence of CO(2). During the course of CO(2) depletion, electron flow to CO(2) was replaced by an equivalent flow to O(2). (c) There was a distinct delay between the cessation of CO(2) uptake and the increase in O(2) uptake. We ascribe this delay to the transient utilization of another electron acceptor-possibly bicarbonate or another bound form of CO(2).