The enzymes that oxidize fatty acids in animal cells are located in the mitochondrial matrix.

Large
FFAs cannot pass through the mitochondria membrane directly, so they need to be transported
which is achieved via three enzymatic reactions of the carnitine shuttle. The first of these
reactions is catalyzed by Acyl-CoA synthetase which promote the general reaction: Fatty acid +
CoA + ATP —> fatty acyl-CoA + AMP + PPi. These fatty acyl-CoAs can then be transported to
matrix of mitochondria where they can undergo oxidation to produce ATP or they can remain in
cytosol to produce membrane lipids. To transport these fatty acyl-CoAs to the matrix, they attach
to the hydroxyl group of carnitine to form the fatty acyl-carnitine—the second reaction of the
carnitine shuttle. The fatty acyl-carnitines then enters the matrix via facilitated diffusion through
the acyl-carnitine/carnitine transporter. Once inside the matrix, the fatty acyl group is
enzymatically transferred from carnitine to the intramitochondrial coenzyme A by carnitine acyl
transferase II, regenerating fatty acyl-CoA and releasing it. The carnitine-mediated entry process
is the rate-limiting step for oxidation of fatty acids in mitochondria. The rate of transfer of long-
chain fatty acyl-CoA in mitochondria is the limiting step of β-oxidation.