SBB 4101 PLANT BIOCHEMISTRY

SUMMARY: PLANT FATTY ACID & ISOPRENOID SYNTHESIS

Done by, SHANMUGAPRAKASHAM S/O SELVAMANI (UK16456) BACHELOR OF SCIENCE (BIOLOGICAL SCIENCES)

PLANT FATTY ACID SYNTHESIS: Beitlenmiller, D. P., Roughan, G. & Ohlrogge, J. B. 1992. Regulation of plant fatty acid biosynthesis. Plant Physiology. 100: 923 930.

The study was done as continues to evaluate factors involved in plant fatty acid biosynthesis as previous study was on the in vivo acyl acyl carier proteins (ACP) involved in fatty acid biosynthesis. In this study, to evaluate role of other factors of substrates and cofactors involved, coenzyme A (CoA), acetyl- and malonyl-CoA were examined. De vovo fatty acid biosynthesis occurs in the plastids and catalyzed by a series of enzymes and mediated by ACP. The precursor substrate of the pathway is acetyl-CoA and melonyl-CoA. From this study, it was found that the major form of CoA in chloroplast and whole leaves is acetyl-CoA and they are also do not differ greatly in light and dark. The transfer of acetyl- or melonyl from CoA to ACP is catalyzed by a transacylase enzyme. Examination of the changes in pool sizes of intermediates in a pathway in this study has provided evidence for the metabolic regulation. Regulatory steps are characterized by increases in their substrate concentration and decreases in product concentration when the flux through the pathway is reduced. Results from this study support the role of acetyl-CoA carboxylase in regulation of plastid fatty acid biosynthesis. Malonyl-CoA levels were undetectable in dark incubated chloroplasts, but rose manyfold in the light and increased further when fatty acid synthesis was stimulated in the light by Triton X-100. The level of acetyl-CoA was reduced concomitantly with the increases in malonyl- CoA. Thus, the changes in both the substrate and product pools of the acetyl-CoA carboxylase reaction provide direct evidence for its role in determining the flux through the fatty acid biosynthetic pathway. Thus, the study provides sufficient evidence of the regulation and role played by acetyl-CoA carboxylase in plant fatty acid biosynthesis.

PLANT ISOPRENOID SYNTHESIS: Calisto, B. M., Perez-gil, J., Bergua, M., Querol-audi, J., Fita, I. & Imperial, S. 2007. Biosynthesis of isoprenoids in plant: structure of the 2c-methyl-D-erithrytol 2,4cyclodiphosphate synthase from Arabidopsis thaliana. Comparison with the bacterial enzymes. Protein Science. 16: 2082 2088.

The study was conducted to construct crystal structure of the 2C-methyl-D-erythritol 2,4cyclodiphosphate (MCS) synthase from Arabidopsis thaliana. Isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP) are the five-carbon precursors of isoprenoids biosynthesis. IPP was identified to be synthesized from acetyl-CoA via mevalonic acid pathway in yeast and animals. There is an alternative pathway for biosynthesis of IPP which was identified later in baceteria. Plants use both pathways. MCS converts 4-diphosphocytidyl 2C-methyl-D-erythritol 2-phosphate (CDP-MEP) to 2Cmethyl D - erythritol 2,4 cyclodiphosphate (MECDP) and cytidine-5-monophosphate (CMP). The first crystal structure from a plant MCS enzyme has been determined, in complex with a CMP molecule. The molecular trimeric organization and the overall structure of subunits, in particular the arrangement of the catalytically essential residues and the active site, are closely related to the ones from the known structures of the bacterial enzymes. Main differences between the plant and the bacterial MCS structures concentrate in the cavity that exists between subunits along the molecular threefold axis and involve residues that are highly conserved among plants. These differences suggest that the binding of isoprenoid diphosphate-like molecules in the cavity of the plant MCS enzymes would not occur, contrary to what had been demonstrated for some bacterial enzymes.