Abstract
The enzymatic activities of the pyruvate dehydrogenase complex (PDC) and acetyl-CoA synthetase (ACS) have been compared in extracts of plastids isolated from spinach leaves and from both green and etiolated pea seedlings. A ll plastid preparations were shown to be capable of synthesizing acetyl-CoA, not only via acetyl-CoA synthetase, but also via the pyruvate dehydroge nase complex, though, with different activities. Both pathways are apparently under metabolic control. Thus, the substrate levels in photosynthetically active spinach chloroplasts appear to favor acetyl-CoA synthesis via ACS (apparent Km for acetate of 0.1 mм) , because calculated stromal pyruvate levels (0.1 m M) appear to limit its formation via the PDC (apparent Km for pyruvate of 0.2-0.3 nм) . In spinach chloroplasts, therefore, the PDC pathway seems to be predominantly involved in providing precursors for branched-chain amino acid biosynthesis (valine, leucine and isoleucine). Acetyl-CoA, synthesized via ACS, may additionally function as an inhibitor of the chloroplast PD C , because, as in mitochondria, relatively low concentrations of the end products NADH and acetyl-CoA strongly inhibit the PD C in chloroplast extracts. On the other hand, comparatively high concentrations of MgATP, a cofactor for ACS, inhibited the PDC complex. The pH optimum of about 8 and the high Mg-requirement distinguishes both enzymes from mitochondrial PDC and reflects an accomodation to stromal conditions in photosynthetically active chloroplasts.
Ethanol represses the expression of methanol-inducible genes via acetyl-CoA synthesis in the yeast Komagataella phaffii
