ABSTRACT: Growth of the green algae Chlamydomonas reinhardtii and Chlorella sp. in batch cultures was investigated in a novel gas-tight photobioreactor, in which CO(2), H(2), and N(2) were titrated into the gas phase to control medium pH, dissolved oxygen partial pressure, and headspace pressure, respectively. The exit gas from the reactor was circulated through a loop of tubing and re-introduced into the culture. CO(2) uptake was estimated from the addition of CO(2) as acidic titrant and O(2) evolution was estimated from titration by H(2), which was used to reduce O(2) over a Pd catalyst. The photosynthetic quotient, PQ, was estimated as the ratio between O(2) evolution and CO(2) up-take rates. NH(4) (+), NO(2) (-), or NO(3) (-) was the final cell density limiting nutrient. Cultures of both algae were, in general, characterised by a nitrogen sufficient growth phase followed by a nitrogen depleted phase in which starch was the major product. The estimated PQ values were dependent on the level of oxidation of the nitrogen source. The PQ was 1 with NH(4) (+) as the nitrogen source and 1.3 when NO(3) (-) was the nitrogen source. In cultures grown on all nitrogen sources, the PQ value approached 1 when the nitrogen source was depleted and starch synthesis became dominant, to further increase towards 1.3 over a period of 3-4 days. This latter increase in PQ, which was indicative of production of reduced compounds like lipids, correlated with a simultaneous increase in the degree of reduction of the biomass. When using the titrations of CO(2) and H(2) into the reactor headspace to estimate the up-take of CO(2), the production of O(2), and the PQ, the rate of biomass production could be followed, the stoichiometrical composition of the produced algal biomass could be estimated, and different growth phases could be identified.
