Fatty Acids and TriglyceridesSynthesizing Fuel Stores

Dianzheng Zhang, Ph.D.

Objectives:
To understand how fatty acids are synthesized
To realize the differences between fatty acids and triglycerides
To appreciate the benefits of fuel storage

Carbohydrates

Liver Lipogenesis

Conditions:
insulin
glucagon
Glucose Fatty Acids
Triglycerides VLDL
(Blood)
VLDL breakdown to Fatty
acids for:
Storage in adipose
Use by muscle, etc.

Concept Map of Fuel Synthesis when insulin, glucagon

References:
Champe & Harvey, Biochemistry, 3rd Ed., Chapter 16 (pp. 179-187)
Marks Basic Medical Biochemistry, 2nd Ed., Chapter 33 (pp. 594ff)

FA & TG Synthesis-1

I.

Fatty Acid Synthesis

A.
OVERVIEW of LIPOGENESIS / FATTY ACID SYNTHESIS:
Why does eating excess carbohydrates result in fat formation??
Converting excess dietary glucose adipose tissue triglycerides
1.

Accumulation of reactants for fatty acid synthesis

What is needed for fat synthesis:

Substrate: Acetyl CoA (in the cytosol)

Energy: ATP
Reducing power: NADPH
From (1) the pentose pathway (see CHO lecture)
(2) the NADP+-linked malic enzyme
Glycerol backbone
Figure 1: Overview of

Lipogenesis
[Marks. Basic Med Biochem. Fig 33.1]

FA synthesis occurs mainly in

the liver
GlucosePyruvateAcetyl CoA
FA synthesisTGs

Figure 2: 1ST STEP:

Conversion of glucose to acetyl CoA. Acetyl CoA cannot move through mitochondrial
membrane. It must be converted to citrate, which can be transported to the cytosol by a
membrane transporter. In the cytosol citrate is converted back to acetyl CoA and OAA by citrate
lyase (also called citrate cleavage enzyme).
[Marks. Basic Med Biochem. Fig 33.7]

FA & TG Synthesis-2

C
Figure 3: Obtaining NADPH for lipogenesis
[Marks. Basic Med Biochem. 2nd Ed. Fig 36.4, p. 671]

Provided by (1) the pentose pathway and (2) the malic enzyme.
Cytosolic malate dehydrogenase and the malic enzyme provide a transhydrogenase
mechanism in the cytosol to transfer hydrogen from NADH to NADPH.

B. Mechanics and enzyme action in Fatty acid synthesis

Stage 1: Acetyl CoA carboxylase
A biotin containing enzyme
The regulated step (see later)

Fig. 4. Acetyl CoA Carboxylase reaction

[Marks Basic Medical Biochemistry, 2nd Ed. Fig. 33.15A, p. 601]

Stage 1: Formation of substrate for Fatty Acid Synthase (malonyl CoA)

Carboxylases always use biotin as a cofactor.

FA & TG Synthesis-3

FA Synthase has 2 armsthe

phosphopantetheine and a
cysteine amino acid. Both
attach to FAs via SH groups.

Stage 2: Fatty acid synthase:

Is a dimer2 identical subunits.

Each subunit is a multifunctional protein

Each subunit contains seven enzyme activities
and an acyl carrier group (often called "acyl carrier protein").
The acyl carrier group contains a phosphopantetheine prosthetic group.
Stage 1: production of
Malonyl CoA

Figure 5: Stage 2: formation of the fatty acyl chain (the fatty acid synthase reactions)
(enzymatic reactions of fatty acid synthase)
[Marks. Basic Med Biochemistry 2nd Ed. Fig 33.15, p. 601]

Rxn
Rxn

Priming step: Covalent attachment of 1st acyl (from acetyl CoA) to the enzyme
(initially onto the phosphopantetheine-SH, then transferred over to the cysteine-SH)
Covalent attachment of malonyl group to the phosphopantetheine of the enzyme
(loading step).
Condensation of the carboxyl carbon of the acetyl group with the methylene carbon of the
malonyl group; CO2 is released; a 4-carbon keto chain is formed.
Reduction, dehydration and reduction stepsformation of a 4-carbon saturated chain.
Shift of the completely reduced chain to the cysteine-SH.
Attachment of another malonyl group to the enzyme. Another condensation occurs and
the cycle repeats to form a 6-carbon saturated acyl chain. The cycle continues to repeat
until a 16-carbon acyl chain (palmitic acid) is formed and is released from the enzyme
complex.

FA & TG Synthesis-4

Stage 3:

ELONGATION of palmitate and

DESATURATION of fatty acids
Fig. 6: Elongation of long chain fatty acids
[Marks. Basic Med Biochem. Fig 33.17]

Acyl CoA Synthetase attaches the CoA to palmitate.

(lipid lecturerequires ATP).
Occurs in the ENDOPLASMIC RETICULUM.
Similar reactions to fatty acid synthesis except that
A fatty acyl CoA is the substrate which condenses
with malonyl CoA.

Fig. 7: Desaturation of fatty acids: creating double bonds

[Marks. Basic Med Biohcem. Fig 33.18]

Occurs in the ENDOPLASMIC RETICULUM.

Uses molecular oxygen.
Two things are oxidizedthe fatty acid and
the NADH.
Human desaturases cannot introduce double bonds
between carbon 9 and the methyl end.
When we need other double bonds, we use essential
fatty acids as precursors (linolenic and linoleic).

FA & TG Synthesis-5

Humans cannot introduce double-bonds between carbon 9 and the methyl end.
However, we can add double bonds to the essential fatty acids that have double bonds
beyond carbon 9.

Fig. 8. Conversion of linoleic acid to arachidonic acid.

[Marks Basic Medical Biochemistry, 2nd Ed. Fig. 33-19 p. 603]

Linoleic acid (18:2, 9,12) is one of the essential fatty acids. It serves as a precursor of
arachidonic acid, which is the source eicosanoids, prostaglandins and leukotrienes.
Linolenic acid (18:3, 9,12,15) also forms eicosanoids.

FA & TG Synthesis-6

C.

Regulation of fatty acid synthesis

Conditions for FA synthesis: when there are lots of reactants:
ATP, Acetyl CoA, NADPH (lots of citrate in TCA cycle)

Figure 8: Regulation of Acetyl CoA carboxylase

(1) Short term regulation (minutes to hours) via Acetyl CoA carboxylase:
The enzyme exists as inactive monomers that polymerize when active.
citrate
Inactive Monomers

Active Polymer
fatty acyl CoA

Covalent Modification
o Glucagon activates cAMP dependent protein kinase A to phosphorylate, inactivate.
o Insulin activates a phosphatase to remove phosphate group, activate.
(2) Long term regulation (days):
Induction (changes in cellular content) of key enzymes:
acetyl CoA carboxylase, fatty acid synthase, citrate lyase, malic enzyme, G6PDH,
and others
Enzyme synthesis is increased (due to insulin and glucagon levels)
If an individual has a good diet over time:
If an individual has a high carbohydrate OR fat free diet:
Enzyme synthesis is decreased
If an individual is fasting OR on a high fat diet:

FA & TG Synthesis-7

(3).

Regulation to prevent a futile cycle between

FA synthesis and breakdown:

A futile cycle between fatty acid synthesis and fatty acid breakdown is avoided because
malonyl CoA (the product of the 1st stage of FA synthesis) inhibits fatty acids
from being transferred to carnitine and entering the mitochondria.

Fig. 9. Inhibition of transport of fatty acids (FA) into mitochondria by malonyl CoA.
[Marks Basic Medical Biochemistry 2nd Ed. Fig. 36.6, p. 672]

Specifically, in the fed state, when malonyl CoA is being formed for FA synthesis, malonyl
CoA inhibits CPTI (carnitine palmitoyltransferase I, or carnitine:acyltransferase I).
[See Lipids lecture for more on CPTI.]

FA & TG Synthesis-8

II.

Triglyceride synthesis

Glycerol-3-phosphate synthesis

Figure 10: Synthesis of glycerol-3- phosphate in liver and adipose tissues.

[Marks. Basic Med Biochem. Fig 33.20]

Glycerol-P-dehydrogenase is present in both

liver and adipose;
Glycerol kinase is present only in liver.

Addition of acyl-CoA chains

to form triacylglycerol

Figure 11: Synthesis of triacylglycerol in liver and

adipose tissues.
[Marks. Basic Med Biochem. Fig 33.20]

Glycerol 3-phosphate is produced from glucose in

both liver and adipose.
It can also be produced directly from glycerol in the
liver, which has glycerol kinase.
Glycerol 3-phosphate is not produced from glycerol in
adipose tissue, which lacks glycerol kinase.
After glycerol 3-phosphate is formed, the steps are
the same in both liver and adipose.

A futile cycle between TG synthesis and TG breakdown is avoided in the adipose

because the adipose must obtain glycerol-3-phosphate from glycolysis; there is
no glycerol kinase found in the adipose.
When HSL (activated by glucagon) cleaves TG to FA + glycerol, the reaction is not
reversible without glycerol kinase, so the FA + glycerol leave the adipose to be
used for energy.
TGs are only synthesized in times of plenty, when glycolysis is increased.

FA & TG Synthesis-9

3.

Formation of VLDL
Triglycerol molecules made by the liver are carried through the blood on
Very Low Density Lipoproteins (VLDL).

Figure 12: Synthesis, processing and

secretion of VLDL.
[Marks. Basic Med Biochem. Fig 33.24]

Proteins (Apo B100) are synthesized on the

rough endoplasmic reticulum (RER).
Triacylglycerols are synthesized on the smooth
endoplasmic reticulum (SER) and/or on FA
synthase in the cytosol.

TG and proteins are packaged in the Golgi

complex to form VLDL.

VLDL are transported to the cell membrane in

secretory vesicles and secreted by exocytosis.
The dots represent VLDL particles. An enlarged
VLDL particle is depicted at the bottom of the
figure.

Fig. 13. Composition of a typical VLDL particle.

The major component is triacylglycerol (TG).
C = cholesterol; CE = cholesterol ester;
PE = phospholipid.
[Marks. Basic Med Biochem. Fig 33.21]
FA & TG Synthesis-10

4.

Transport and Storage of Triglycerides

Transport of triacylglycerols from liver (site of synthesis) to
The adipose tissue (site of storage)

Figure 14: Transport of TG in blood:

1. Glucose enters the liver and is
converted to fatty acids and
glycerol-3-P.
2. Triglycerides are formed from FACoAs and glycerol-3-P.
3. Triglycerides are incorporated into
very-low-density lipoproteins
(VLDL).
4. VLDL goes into the blood and
carried to adipose tissue.
5. Triglycerides in the VLDL are
hydrolyzed by lipoprotein lipase
(LPL) and the released FAs are
taken into adipose tissue.
6. In the adipocytes, the FAs are
resynthesized into triglycerides for
storage.

B.

Conversion of excess dietary fat to adipose tissue triglycerides

Figure 15: Dietary lipid digestion and

storage:
1. Dietary lipids are digested and
absorbed into intestinal cells.
2. Absorbed lipids are incorporated into
chylomicrons in intestinal cells.
3. Chylomicrons enter the blood and
carry the dietary lipids to adipose
tissue.
4. Triglycerides in the chylomicrons are
hydrolyzed by lipoprotein lipase
(LPL), and the released fatty acids
are taken into adipose tissue.
5. Fatty acids are incorporated into
storage triglycerides.

FA & TG Synthesis-11

Accumulation of triglycerides in adipose tissue

TG storage in the adipocyte

Figure 16: Conversion of fatty acids (FA) from triacylglycerols (TG) in

chylomicrons and VLDL to the TG stored in adipose tissue
[Marks Basic Med Biochemistry 2nd Ed. Fig 36-7, p. 673]

Insulin stimulates the transport of glucose into adipose cells.

Glucose provides the glycerol-3-phosphate for TG synthesis.

Insulin stimulates the synthesis and secretion of lipoprotein lipase (LPL).

ApoCII (obtained from HDL in the blood) activates LPL.

ApoCII is present on both the VLDL and chylomicrons.

FA & TG Synthesis-12