The Step of Fatty Acid Synthesis Fatty acid synthesis from acetyl CoA occurs in the cytoplasm.

Mitochondrial acetyl CoA is unable to diffuse through the inner membrane into the cytoplasm. A shuttle system transports the carbons of acetyl CoA disguised as citrate through the membranes. In the mitochondria, acetyl CoA reacts with oxaloacetate to produce citrate (citrate synthase). Citrate is transported by the integral membrane protein, tricarboxylate translocase, into cytoplasm, where it is c leaved to acetyl CoA and oxaloacetate (citrate lyase). Oxaloacetate is eventually returned to the mitochondria, but acetyl CoA in the cytoplasm is now available for fatty acid synthesis. The final product of the fatty acid synthase complex is palmitate. Palmitate synthesis begins at the terminal methyl end and proceeds toward the carboxylate (Figure 18.16). The first two carbons of the palmitate chain (carb ons 15 and 16) come directly from acetyl CoA. All other carbons come originally from acetyl CoA but must be activated as malonyl CoA. Acetyl-CoA carboxylase, a biotinrequiring enzyme, catalyzes the formation of malonyl CoA:

A positive modulator for the allosteric enzyme is citrate. The enzyme is inhibited by pahttitoyl CoA, the end product of the fatty acid synthesis system.

From this point on, all intermediates are linked to a small molecular weight pro tein called acyl carrier protein (ACP-SH), which is a component of fatty acid synthase. ACP has properties similar to those of CoASH.

The vitamin pantothenic acid is a component of both activating groups. The intermediates for fatty acid synthesis are linked via a thioester bond to the SH group at one end of the ACP. To prepare for fatty acid synthesis, one acetyl CoA and one malonyl CoA must be brought into the synthase complex. They are attached to SH groups, one on an enzyme, - ketoacyl-ACP synthase (K-SH), and one on ACP (malonyl CoA). The reactions consist of a spiral that is the chemical reverse of oxidation: (1) formation of a carbon-carbon single bond, (2) reduction of a keto group, (3) dehydration to form a carbon-carbon double bond, and (4) reduction of the double bond to form the saturated fatty acid chain.

One round through the fatty acid synthase spiral has produced a four-carbon carboxyl group (butyryl) linked to ACP. In the second round, two more carbons are added from malonyl ACP to the butyryl unit to make a C6 -keto intermediate that goes through reduction, dehydration, and reduction. The two new carbons become numbers 11 and 12 in the palmitate product. A total of seven turns of this set of four reaction result in palmytoylACP.

Biosynthesis of Unsaturated Fatty Acids Unsaturated fatty acids are synthesized in the endoplasmic reticulum by enzymes called fatty acyl-CoA desaturases. These enzymes catalyze unique oxidationreduction reactions. The dehydrogenation of stearic acid (18:0) to oleic acid (18:9) is an example: stearoyl CoA + NADPH + H + + O 2 CoA and two from NADPH. Regulation of Fatty Acid Synthesis The two major processes of oxidation and synthesis must be coordinated so they do not occur simultaneously. If both processes were to occur at the same time, a wasteful substrate cycle would develop. The metabolism of fatty acids is controlled by the availability of the nutrients, carbohydrates and fatty acids. If glucose is abundant, high levels of citrate are produced. Citrate is a positive modulator for acetyl-CoA carboxylase, which catalyzes the formation of malonyl CoA. the rate-limiting step in fatty acid synthesis. With increasing citrate, fatty acid synthesis is stimulated. Beta oxidation of fatty acids is inhibited by the malonyl CoA produced. It blocks the action of carnitine acyltransferase I, an enzyme responsible for transport of fatty acid CoA esters into the mitochondrial matrix, where they are oxidized. The rate of fatty acid synthesis is also regulated by the end product, palmitoyl CoA, which inhibits acetyl-CoA carboxylase. In summary, with abundant glucose, fatty acid oxidation is turned off and fatty acid synthesis is active, but regulated by levels of palmitoyl CoA. During fasting or when glucose and glycogen levels become low, the hormones epinephrine and glucagon stimulate hormone-sensitive lipase to release free fatty acids from storage. Lowered glucose levels result in less citrate, therefore, stimulation of acetyl-CoA carboxylase is removed. oleoyl CoA + NADP + + 2 H 2 O

Molecular oxygen as a substrate accepts four electrons in this reaction, two from the stearoyl

Ch ol es terol Cholesterol may be one of the most dreaded chemicals in nature. Scientific evidence certainly supports a "bad side" to cholesterol. However, we hear little about its "good side." This molecule is essential to the life of all animals, indeed, it is a molecule we must live with. Cholesterol plays several roles in biochemistry: 1) it is an essential component of animal membranes, where it regulates fluidity, and 2) it is a precursor of many important biomolecules, including the steroid hormones, bile Cholesterol, a water-insoluble molecule, is very difficult for organisms to dissolve and transport.