13C NMR studies of acetate metabolism during sporulation of Saccharomyces cerevisiae

Abstract

The sporulation of Saccharomyces cerevisiae in the presence of [2-13C]acetate was studied by 13C NMR spectroscopy. The fate of 13C label was analyzed in vivo and in cell extracts. During the first 4 hr of sporulation the major metabolite produced from [2-13C]acetate utilization was glutamate. From the labeling pattern observed it is concluded that both the tricarboxylic acid cycle and the glyoxylate cycle are operating. After about 4 hr trehalose is made. Comparison of the doublet/singlet ratios for C-1,1(1) and C-6,6(1) of trehalose shows a steady drop in the ratio of C-1, C-2-coupled species over trehalose labeled only at C-1 in the C-1, 2 segment of the molecule. The negative correlation of this ratio with that for the C-5, 6 segment indicates a cycling of glucose through the hexose monophosphate shunt. Subsequently fatty acid biosynthesis commences. Large amounts of saturated fatty acid were made. There were conspicuous differences observed in the metabolism of [2-13C]acetate between sporulating and vegetatively growing cells.