Fig 3.

Time course of sPPase activity (A) and growth curves (B) of strains YPC-1 and W303-1A on different carbon sources. Two milliliters of stationary cultures of W303-1A and YPC-1 grown on galactose were used to inoculate flasks containing 200 ml of selective medium and cultured with agitation at 30°C. Specific PPase activity was assayed at different times in crude soluble extracts of mutant YPC-1 and strain W303-1A grown on galactose (hatched columns) or glucose (black columns). Activity values are means ± SEM obtained in three independent experiments. (Inset) Analysis by Western blot of crude soluble extracts obtained from YPC-1 cells at the beginning of inoculation and after 48 h of growth on galactose and glucose using an antibody against the homologous chloroplastic sPPase protein of the microalga C. reinhardtii. Growth was monitored by measuring absorbances at 660 nm. After 48 h (indicated by an arrow), 2 ml of stationary cultures were inoculated into 200 ml of fresh medium, and growth was followed for further 48 h. ○, W303-1A in galactose; •, W303-1A in glucose; □, YPC-1 in galactose; ▪, YPC-1 in glucose.