Kinetic properties of phosphoenolpyruvate carboxylase from c(3), c(4), and c(3)-c(4) intermediate species of flaveria (asteraceae).

Flaveria cronquistii (C(3)), F. chloraefolia (C(3)-C(4)), F. floridana (C(3)-C(4)), F. pubescens (C(3)-C(4)), F. anomala (C(3)-C(4)), F. linearis (C(3)-C(4)), F. brownii (C(4)), F. palmeri (C(4)), F. trinervia (C(4)) and F. australasica (C(4)), comprising 10 out of the 21 known species of the genus Flaveria (Asteraceae), were included in a comparative study of the kinetic and regulatory properties of green leaf phosphoenolpyruvate (PEP) carboxylase. At least three kinetically distinct enzyme-forms were identified on the basis of their affinities for PEP and the degree of allosterism with respect to this substrate. The kinetic properties of PEP carboxylase of most of the species seemingly were modified in vivo depending on the growth conditions of the plants. K(m)(PEP(free))-values of the enzyme from the five C(3)-C(4) intermediate species ranged from 6 micromolar (F. chloraefolia, low light-grown) to 38 micromolar (F. pubescens, high light-grown). In contrast, the K(m) for PEP of PEP carboxylase from the C(3) species F. cronquistii (13 micromolar) apparently was not influenced by growth conditions. The response of the enzyme from the C(3) and C(3)-C(4) species was hyperbolic in all cases. A second isoform with a lower affinity for PEP (88-100 micromolar), but also hyperbolic kinetics was found in the C(4) species F. brownii, whereas in the three other C(4) species examined a PEP carboxylase with a still lower affinity for PEP (187-221 micromolar) and sigmoidal kinetics was present. These isozyme-related kinetic data were supported by analyses of the elution behavior of the enzyme during anion-exchange chromatography on DEAE-Trisacryl M. The results are discussed with respect to the evolution of C(4) photosynthesis in the Flaveria genus.