CARBON SOURCE DEPENDENCE OF THE RATIO OF δ-AMINOLEVULINIC ACID BIOSYNTHESIS VIA THE C5 AND SHEMIN PATHWAYS IN EUGLENA...

The ratio of two biosynthetic pathways was estimated, the C5 and Shemin pathways, to δ-aminolevulinic acid (ALA, a biosynthetic intermediate of tetrapyrrole) from the (13) C-enrichment ratios ((13) C-ER) at the carbon atoms of chl a (after conversion to methyl pheophorbide a) biosynthesized by Euglena gracilis G. A. Klebs when l-[3-(13) C]alanine was used as a carbon source. On the basis of these estimations, we confirmed that ALA was efficiently biosynthesized via both the C5 and Shemin pathways in the plastids of E. gracilis, and we determined that the ratio of ALA biosynthesis via the Shemin pathway was increased in the ratio of 14%-67%, compared with that in our previous d-[1-(13) C]glucose feeding experiment (Iida et al. 2002). This carbon source dependence of the contributions of the two biosynthetic pathways might be related to activation of gluconeogenesis by the amino acid substrate. The methoxy carbon of the methoxycarbonyl group at C-13(2) of chl a was labeled with the (13) C-carbon of l-[methyl-(13) C]methionine derived from l-[3-(13) C]alanine via [2-(13) C]acetyl coenzyme A (CoA), through the atypical tricarboxylic acid (TCA) cycle, gluconeogenesis, and l-[3-(13) C]serine. The phytyl moiety of chl a was also labeled on C-P2, C-P3(1) , C-P4, C-P6, C-P7(1) , C-P8, C-P10, C-P11(1) , C-P12, C-P14, C-P15(1) , and C-P16 from (13) C-isoprene (2-[1,2-methyl,3-(13) C3 ]methyl-1,3-butadiene) generated from l-[3-(13) C]alanine via [2-(13) C]acetyl CoA.