Fatty acyl-CoA derived from activation of fatty acid cannot enter the inner mitochondrial membrane directly. Instead, the acyl portion is treated with carnitine to give acyl-carnitine. The transesterification reaction occurs by the action of Carnitine palmitoyl transferases I, which transfers the acyl group to carnitine.
The acyl-carnitine is transferred to the inner mitochondrial matrix by the carnitine carrier protein. Then again, the same Carnitine palmitoyl transferases I present in the inner surface of the mitochondrial region liberate the free carnitine from acyl-carnitine. In this way, cytosolic acyl groups can enter the mitochondrial membrane while free carnitine will return to the cytosol.