Fatty Acid Synthesis

Prepared By: Divine Mary C. Mamalayan III-A BSBT

Fatty acid biosynthesis

 Fatty acids can be synthesized from acetyl-CoA. This is the major way of utilizing excess dietary carbohydrates. Fatty acid synthesis occurs mainly in the fat tissue and the liver. Synthesis and degradation of fatty acids are similar in that they involve a cycle of reactions that changes the length of the substrate by two carbon atoms at a time.

A three carbon intermediate, malonyl-CoA, initiates fatty acid synthesis

Glycerol-P

FA biosynthesis and breakdown occur by different pathways and take place in different parts of the cell.

Glucose Triacylglycerol

Fatty acyl CoA

Pyruvate Acetyl CoA TCA cycle

Malonyl CoA

The carboxylation of acetyl-CoA yields malonyl-CoA

O Acetyl-CoA CH3-C-S-CoA
Acetyl CoA carboxylase

O OOC-CH2-C-S-CoA

HCO3-

Malonyl-CoA

Assembly of a long chain fatty acid

Once malonyl-CoA is synthesized, long carbon FA chains may be assembled in a repeating four-step sequence. With each passage through the cycle the fatty acyl chain is extended by two carbons. When the chain reaches 16 carbons, the product palmitate (16:0) leaves the cycle.

The first round of FA biosynthesis

To initiate FA biosynthesis, malonyl and acetyl groups are activated on to the enzyme fatty acid synthase.
Malony-CoA

+
Acetyl-CoA

H H

Step 1.

Condensation of an activated acyl group and two carbons derived from malonyl-CoA

Step 2.
The -keto group is reduced to an alcohol by NADPH

Step 3.
The elimination of water creates a double bond.

Step 4.
The double bond is reduced to form the correspondin g saturated fatty acyl group.

Repetition of these four steps leads to fatty acid synthesis

When reaches 16 carbons, the product leaves the cycle. All the reactions in the synthetic process are catalyzed by a multienzyme complex, fatty acid synthase.

A more detailed look at fatty acid synthase

Fatty acyl synthase contains six enzymatic activities

Each segment of the disk represents one of the six enzymatic activities of the complex. At the center is the ACP acyl carrier protein - with its phosphopantetheine arm ending in SH.

The function of the prosthetic group of the ACP

Serve as a flexible arm, tethering the growing fatty acyl chain to the surface of the synthase complex Carrying the reaction intermediates from one enzyme active

Activation of acetyl and malonyl groups

Before Steps 1-4, the two thiol groups on the enzyme complex must be charged with the correct acyl groups.

The activation of the acetyl group

The acetyl group from acetyl-CoA is transferred to the Cys-SH group of the -ketoacyl ACP synthase. This reaction is catalyzed by acetylCoA transacetylase.

The activation of the malonyl group

Transfer of the malonyl group to the SH group of the ACP is catalyzed by malonyl-CoA ACP transferase. The charged acetyl and malonyl groups are now in close proximity to

Step 1.
Condensation of the activated acetyl and malonyl groups to form acetoacetylACP, catalyzed by ketoacyl-ACP synthase.

Step 2.
Reduction. The acetoacetyl-ACP is reduced to -hydroxybutyryl-ACP, catalyzed by ketoacyl-ACP reductase (needs NADPH + H+)

Step 3.
Dehydration to yield a double bond in the product, trans-2-butenoyl-ACP, catalyzed by hydroxyacyl-ACP dehydratase.

Step 4.
Reduction of the double bond to form butyryl-ACP, catalyzed by enoylreductase. Another NADPH dependent reaction.

The growing chain is transferred from the acyl carrier protein

This reaction makes way for the next incoming malonyl group. The enzyme involved is acetyl-CoA transacetylas e.

Beginning of the second round of the FA synthesis cycle

The butyryl group is on the Cys-SH group. The incoming malonyl group is first attached to ACP. In the condensation step, the entire butyryl group is exchanged for the carboxyl group on the malonyl residue.

The result of fatty acyl synthase activity

Seven cycles of condensation and reduction produce the 16-carbon saturated palmitoyl group, still bound to ACP. Chain elongation usually stops at this point, and free palmitate is released from the ACP molecule by hydrolytic activity in the synthase complex. Smaller amounts of longer fatty acids such as stearate (18:0) are also formed.

The overall reaction for the synthesis of palmitate from acetylCoA can be considered in two parts.

Part 1.

First, the formation of seven malonylCoA molecules: 7Acetyl-CoA + 7CO2 + 7ATP 7malonyl-CoA + 7ADP + 7Pi

Part 2.
Then the seven cycles of condensation and reduction

Acetyl-CoA + 7malonyl-CoA + 14NADPH + 14H+ palmitate + 7CO2 + 8CoA + 14NADP+ + 6H2O The biosynthesis of FAs requires acetyl-CoA and the input of energy in the form of ATP and reducing power of NADPH.

Location of FA synthesis
FA synthase complex is found exclusively in the cytosol. The location segregates synthetic processes from degradative reactions.

In hepatocytes:
The [NADPH]/[NAD+] ratio is very high (~75) in the cytosol, furnishing a strongly reducing environment for the reductive synthesis of fatty acids and other biomolecules.

Fatty acid synthesis requires considerable amounts of NADPH + H+

Acetyl-CoA + 7malonyl-CoA + 14NADPH + 14H+ palmitate + 7CO2 + 8CoA + 14NADP+ + 6H2O

In hepatocytes and adipocytes, cytosolic NADPH is largely generated by the malic enzyme and by the pentose phosphate pathway.

1. The malic enzyme

The pyruvate produced in the reaction reenters the mitochondrion.

2. The pentose phosphate pathway

In hepatocytes and the mammary gland of lactating animals, the NADPH is supplied primarily by the pentose phosphate pathway.

Fatty acid synthesis requires considerable amounts of acetylCoA

7Acetyl-CoA + 7CO2 + 7ATP + 7Pi 7malonyl-CoA + 7ADP

Nearly all acetyl-CoA used in fatty acid synthesis is formedCytosol site of acetate utilization in mitochondria from pyruvate oxidation.
Mitochondria site of acetate manufacture

Acetate is shuttled out of mitochondria as citrate

The mitochondrial inner membrane is impermeable to acetyl-CoA Intramitochondrial acetyl-CoA first reacts with oxaloacetate to form citrate, in the TCA cycle catalyzed by citrate synthase.

 Citrate then passes into the cytosol through the mitochondrial inner membrane on the citrate transporter. In the cytosol, citrate is cleaved by citrate lyase regenerating acetyl-CoA.

 The other product --oxaloacetate cannot return to the mitochondrial matrix directly. Instead, oxaloacetate is reduced to malate

 Malate returns to the mitochondrial matrix on the malate--ketoglutarate transporter in exchange for citrate.

Regulation of fatty acid synthesis

When a cell has more energy, the excess is generally converted to FAs and stored as lipids such as triacylglycerol. The reaction catalyzed by acetylCoA carboxylase is the rate limiting step in the biosynthesis of fatty acids.

The carboxylation of acetylCoA yields malonyl-CoA

O Acetyl-CoA CH3-C-S-CoA

O OOC-CH2-C-S-CoA

HCO3-

Malonyl-CoA

Regulation of acetyl-CoA carboxylase(1)

Palmitoyl-CoA acts as a feedback inhibitor of the enzyme, and citrate is an activator. When there is an increase in mitochondrial acetyl-CoA and ATP, citrate is transported out of mitochondria, Citrate becomes both the precursor of cytosolic acetyl-CoA and a signal for the activation of acetylCoA carboxylase.

Regulation of acetyl-CoA carboxylase (2)

Regulation of acetyl-CoA carboxylase (3)

Additionally, these pathways are regulated at the level of gene expression. For example, when animals ingest an excess of certain polyunsaturated fatty acids, the expression of genes encoding a wide range of lipogenic enzymes in the liver is suppressed.

Additional modification to the newly synthesized fatty acid

Extended to form longer fatty acids Converted to monounsaturated and polyunsaturated fatty acids

Fatty Acid elongation

Palmitate in animal cells is the precursor of other long-chained FAs. By further additions of acetyl groups, through the action of FA elongation systems present in the smooth endoplasmic reticulum and the mitochondria.

The desaturation of FAs

Palmitate and stearate serve as precursors of the two most common monosaturated fatty acids of animal cells: palmitoleate (16:1D9), and oleate (18:1D9). The double bond is introduced by fatty acyl-CoA desaturase in the smooth endoplasmic reticulum.

Essential fatty acids

Mammalian hepatocytes readily introduce double bonds at the 9 position of FAs but cannot between C-10 and the methylterminal end. Linoleate, 18:29,12 and linolenate 18:39,12,15 cannot be synthesized by mammals, but

The fate of fatty acids

Most of the FAs synthesized or ingested by an organism have one of two fates:

incorporated into triacylglycerols for the storage of metabolic energy incorporation into the phospholipid components of membranes.

The formation of phosphatidic acid

Fatty acyl groups are first activated by formation of fatty acyl-CoA molecules. then transferred to ester linkage with Lglycerol 3phosphate.

Phosphatidic acid may be converted to triacylglycerols or phospholipids Triacylglycerols and phosholipids are both synthesized from phosphatidic acid

Partitioning of the fates of fatty acids

Depends on the needs of the organism: During rapid growth, synthesis of new membranes requires membrane phospholipid synthesis Organisms that have a plentiful supply of food but are not actively growing shunt most of their fatty acids into storage fats.

Genetic Disorder.
What is malonyl-CoA decarboxylase deficiency? Malonyl-CoA decarboxylase deficiency is a condition that prevents the body from converting certain fats to energy. The signs and symptoms of this disorder typically appear in early childhood. Almost all affected children have delayed development. Additional signs and symptoms can include weak muscle tone (hypotonia), seizures, diarrhea, vomiting, and low blood sugar (hypoglycemia). A heart condition called cardiomyopathy, which weakens and enlarges the heart muscle, is another common feature of malonyl-CoA decarboxylase deficiency.

 How common is malonyl-CoA decarboxylase deficiency? This condition is very rare; fewer than 30 cases have been reported. What genes are related to malonyl-CoA decarboxylase deficiency? Mutations in the MLYCD gene cause malonyl-CoA decarboxylase deficiency. The MLYCD gene provides instructions for making an enzyme called malonyl-CoA decarboxylase. Within cells, this enzyme helps regulate the formation and breakdown of a group of fats called fatty acids. Many tissues, including the heart muscle, use fatty acids as a major source of energy. Mutations in the MLYCD gene reduce or eliminate the function of malonylCoA decarboxylase. A shortage of this enzyme disrupts the normal balance of fatty acid formation and breakdown in the body. As a result, fatty acids cannot be converted to energy, which can lead to characteristic features of this disorder including low blood sugar and cardiomyopathy. Byproducts of fatty acid processing build up in tissues, which also contributes to the signs and symptoms of malonyl-CoA decarboxylase deficiency.

Summary of lipid metabolism

FA biosynthesis requires malonyl-CoA formation The long carbon chains of FA acids are assembled in a repeating four-step sequence catalyzed by the multifunctional enzyme fatty acid synthase. With each passage through the cycle, the fatty acyl chain is extended by two carbons

 Cytosolic NADPH is largely generated by the malic enzyme and by the pentose phosphate pathway. FA biosynthesis occurs in the cytosol FA biosynthesis is regulated by the activity of acetyl-CoA carboxylase Synthesized FA are either stored as TG or made into membrane lipids

Thanks for Listening!