Partial characterization of a malonyl-CoA-sensitive carnitine O-palmitoyltransferase I from Macrobrachium borellii...

The shuttle system that mediates the transport of fatty acids across the mitochondrial membrane in invertebrates has received little attention. Carnitine O-palmitoyltransferase I (EC 2.3.1.21; CPT I) is a key component of this system that in vertebrates controls long-chain fatty acid beta-oxidation. To gain knowledge on the acyltransferases in aquatic arthropods, physical, kinetic, regulatory and immunological properties of CPT of the midgut gland mitochondria of Macrobrachium borellii were assayed. CPT I optimum conditions were 34 degrees C and pH=8.0. Kinetic analysis revealed a Km for carnitine of 2180+/-281 microM and a Km for palmitoyl-CoA of 98.9+/-8.9 microM, while V(max) were 56.5+/-6.6 and 36.7+/-4.8 nmol min(-1) mg protein(-1), respectively. A Hill coefficient, n~1, indicate a Michaelis-Menten behavior. The CPT I activity was sensitive to regulation by malonyl-CoA, with an IC(50) of 25.2 microM. Electrophoretic and immunological analyses showed that a 66 kDa protein with an isoelectric point of 5.1 cross-reacted with both rat liver and muscle-liver anti CPT I polyclonal antibodies, suggesting antigenic similarity with the rat enzymes. Although CPT I displayed kinetic differences with insect and vertebrates, prawn showed a high capacity for energy generation through beta-oxidation of long-chain fatty acids.