Chapter 19 1 Takusagawa’s Note©

Chapter 19: Lipid Metabolism

Triacylglycerols = triglycerides = fats
- Triacylglycerols are the major biological energy source, which are water insoluble.
Glycogen is also biological energy source, but it takes a large space to store since glycogen is
heavily hydrated.
- Typical triacylglycerol is 1-palmitoyl-2,3-dioleoyl-glycerol.
O
H2C1 O C 16
O 9
HC2 O C
18
O 9
H2C3 O
18

1. Dietary fat is breakdown to two fatty acids and monoacylglycerol by pancreatic lipase.
2. The fatty acids and monoacylglycerol are absorbed into small intestinal enterocyte cells.
3. In the cell, fatty acids and monoacylglycerol are reattached to form triacylglycerol.
4. Triacylglycerols are wrapped by proteins to form chylomicrons.
5. Chylomicrons enter into lymph duct, and then blood stream.
Dietary fat
(triacylglycerol)

Pancreatic Enterocyte cells of intestinal wall

Lipase
Lymph
2 fatty acids 2 fatty acids 2 CoA duct

+
2 fatty acyl CoA
Monoacylglycerol Monoacyl-
glycerol

Triacylglycerol
Intestinal Lumen Chylo- Chylo-
Protein
microns microns

Transport of chylomicrons and cholesterol in human is shown in Fig. 11-53.

1
 Chapter 19 2 Takusagawa’s Note©

Fatty acids in phospholipids are hydrolytically cleaved by various phospholipase as shown in

Fig. 23-2.

Phospholipase A2

2
 Chapter 19 3 Takusagawa’s Note©

Phospholipase A2
- As shown in the previous page, phospholipase A2 cleaves C-2O bond of phospholipid.
- The enzyme contains a Ca2+ and has a His-48 - Asp-99 “catalytic diad”.
- His-48 abstracts a proton from a water molecule in the active site.
- The activated HO- nucleophilic attacks on the 2C (carbonyl carbon) of phospholipid.
- The Ca2+ stabilizes the oxyanion transition state.

3
 Chapter 19 4 Takusagawa’s Note©

Metabolism of triacylglycerol in adipose cells

- When certain hormones, such as epinephrine and norepinephrine, bind to their receptors in
adipose tissue, adenylate cyclase is activated.

Hormone-receptor Plasma membrane

interaction

Adenylate cyclase
PPi Adipose cell
ATP
cAMP

Protein kinase Protein kinase

(inactive) (active)
ADP
2+
ATP Mg Triacylglycerol
Triacylglycerol Triacylglycerol
lipase (inactive) lipase (active) Fatty acid
2+
Pi Mg P
Diacylglycerol

Fatty acid
Monoacylglycerol

Fatty acid
Glycerol

The free fatty acids are released into bloodstream. The maximum solubility of fatty acid in
blood is relatively low (~10-6 M). Thus, the fatty acids are bind to albumin that is soluble
protein and comprises about half of the blood serum protein.

4
 Chapter 19 5 Takusagawa’s Note©

Fatty acid oxidation

- This is called β-oxidation since the β-carbon of fatty acid is oxidized.
H H O
H3C (CH2)n C Cα C S CoA
β
H H
- The β-oxidation leads acetyl-CoA and 2-carbons less fatty acid at a time.
O O O
HSCoA
H3C (CH2)n C S CoA H3C (CH2)n-2 C S CoA + H3C C S CoA
Fatty acid (Cn+2)-CoA Fatty acid (Cn)-CoA Acetyl-CoA

- CoA is attached to fatty acid by using ATP hydrolysis energy, and acyl-CoA is yielded.
Fatty acid + CoA + ATP → acyl-CoA + AMP + PPi (→ 2Pi)
- In this reaction, the negatively charged carboxyl oxygen (⎯O-) of fatty acid attacks the α-
phosphate of ATP, and the pyrophosphate is cleaved from ATP. This reaction is slightly
endergonic. The subsequent reaction (pyrophosphate hydrolysis) pulls the previous
endergonic reaction, since the hydrolysis of the pyrophosphate is relatively large exergonic
reaction (ΔG = -33.5 kJ/mol).

Endergonic

Exergonic

5
 Chapter 19 6 Takusagawa’s Note©

- Why does nature convert fatty acids to acyl-CoA in order to metabolize them?
- Answer will be:
O O O
CH2 C OH CH2 C Cl because CH2 C OH can takes

less active more active

-
O O
+
R C O R' R C O R' resonance forms.

- However, if the O-R’ is replaced with S-R’, the above resonance is not as important in the
thiol ester. The thiol ester in acyl-CoA behaves like a ketons (no resonance forms, thus less
stable). O
R C SCoA

- Therefore the hydrogens on α-carbon are more acidic. Thus, they can react with base (-B:).
O O
B + H CH2 C SCoA BH + CH2 C SCoA
δ- δ+
- Therefore, attachment of CoA to the fatty acid carboxyl group weakens the Cα-Cβ bond of
acyl group.

Transport of acyl-CoA produced in cytosol

- Acyl-CoA is made in cytoplasm, but β-oxidation is carried out in mitochondrion. Thus,
Acyl-CoA must be transported into mitochondrion. However, Acyl-CoA itself does not
have a specific transport protein. Thus, a special carrier system is used.
- Initially acyl-CoA reacts with carnitine, and acyl-carnitine and CoA are produced.

6
 Chapter 19 7 Takusagawa’s Note©

- The acyl-carnitine is transported through the mitochondrial membrane at the carnitine carrier
protein to mitochondrion. The acyl-carnitine reacts with CoA in mitochondrion, and
produce acyl-CoA and carnitine.

β-oxidation is 4 steps with 4 enzymes (Fig. 23-10).

1. Desaturation between α and β carbon by using FAD to FADH2 reduction, and yield a trans
Δ2 compound. Enzyme: acyl-CoA dehydrogenase
FAD FADH2
H H O H O
H3C (CH2)n C Cα C S CoA H3C (CH2)n C Cα C S CoA
β β
H H H

2. Hydration with H2O to 3 carbon to yield 3-L-hydroxyacyl-CoA. Enzyme: enoyl-CoA

hydratase.
H2O
H O H H O
H3C (CH2)n C Cα C S CoA H3C (CH2)n C C C S CoA
β
H OH H

3. Oxidation by using NAD+ to NADH, and yield β-ketoacyl-CoA. Enzyme: 3-L-hydroxyacyl-

CoA dehydrogenase.
+ NADH
H H O NAD
O H O
H3C (CH2)n C C C S CoA H3C (CH2)n C C C S CoA
OH H H

4. Cleavage by CoA to produce Acetyl-CoA and acyl-CoA with n-2 carbons. Enzyme: β-
ketoacyl-CoA thiolase.
O H O CoAH
H H O O
H3C (CH2)n C C C S CoA H3C (CH2)n-2 C C C S CoA + H3C C S CoA
H H H

7
 Chapter 19 8 Takusagawa’s Note©

- Overall β-oxidation reaction is shown below.

ETF = Electron-transfer flavoprotein

8
 Chapter 19 9 Takusagawa’s Note©

Energetic of fatty acid oxidation

1. Saturated even number carbon atoms (n = even, such as 14, 16 and 18)
Cn fatty acid + CoASH + 2ATP → (Cn fatty acid)-CoA
FADH2

2 6 4 2
6 4 1
1 SCoA SCoA
C 5
C
5 3 3
O O
Acetyl-CoA
FADH2 + NADH
6 4
3 SCoA 6 4
3
C SCoA
5
5
C
O
O
+
1: (Cn fatty acid)-CoA + FAD + H2O + NAD + CoASH →
(Cn-2 fatty acid)-CoA + acetyl-CoA + FADH2 + NADH
↓
↓
n/2-1: (C4 fatty acid)-CoA + FAD + H2O + NAD+ + CoASH →
(C2 fatty acid)-CoA + acetyl-CoA + FADH2 + NADH
From one fatty acid, (n/2 - 1) NADH and FADH2
(n/2) Acetyl-CoA
For example, C16 fatty acid (palmitic acid):
16/2 - 1 = 7 NADH = 21 ATP
7 FADH2 = 14 ATP
16/2 = 8 Acetyl-CoA = 96 ATP*
* 3 NADH, 1 FADH2, 1 ATP = 12 ATP/acetyl-CoA
Total = 131 ATP - 2* = 129 ATP (8.1 ATP/C)
* 2 ATPs are utilized to attach CoA to fatty acid
Note: one glucose produces 38 ATPs (6.3 ATP/C).

2. Unsaturated fatty acids.

Double bonds occur most often Δ9 & higher, and occur every three carbons, such as Δ9, 12, 15.
- Example, Δ9 fatty acid:
Δ9 ⎯ ⎯⎯⎯⎯⎯ ⎯→ Δ7 ⎯ ⎯⎯⎯⎯⎯ ⎯→ Δ5 ⎯ ⎯⎯⎯⎯⎯ ⎯→ Δ3 (cis double bond)
Acetyl − CoA Acetyl − CoA Acetyl − CoA
- Cis double bond cannot be cleaved by β-oxidation (i.e., cannot hydrate (H2O addition)).
- Δ3 must be transferred to Δ2 by enoyl-CoA isomerase.
- The acetyl-CoA dehydrogenase step (step-1) is skipped (i.e., no FADH2 production).

O enol-CoA isomerase 6 4 2
1
6 4 3 SCoA
C C
2
1 SCoA No FADH2 5 3
5 O

9
 Chapter 19 10 Takusagawa’s Note©

- Two double bonds case, such as Δ9, 12.

Δ9, 12 ⎯ ⎯⎯⎯⎯⎯ ⎯→ Δ7,10 ⎯ ⎯⎯⎯⎯⎯ ⎯→ Δ5, 8 ⎯ ⎯⎯⎯⎯⎯ ⎯→ Δ3, 6
Acetyl − CoA Acetyl − CoA Acetyl − CoA
- After isomerization and one cycle β-oxidation, Δ2, 4 (trans Δ2, cis Δ4) is yielded.
- Δ2, 4 is a poor substrate, and must be reduced to Δ3 by 2,4-dienoyl-CoA reductase.
- This process requires one NADPH.
The Δ3 must be transferred to Δ2 by 3,2-enoyl-CoA isomerase.

7 6 O enol-CoA isomerase 7 6 4 2
4 3 1 SCoA
C C
2
1 SCoA 8 5 3
8 5 O
Acetyl-CoA
FADH2 + NADH
7 6 4
3 SCoA 7 6 4
SCoA
3
C
8 5 C
8 5
O O
NADPH
Acetyl-CoA
+ NADH
8 6 4 8 6
3 SCoA 5 SCoA
C C
7 5 7
O O

3. Odd carbon fatty acids (Cn, n = odd number)

10
 Chapter 19 11 Takusagawa’s Note©

At final step,
HSCoA
O O O O
CH3CH2 C CH2 C SCoA CH3CH2 C SCoA + CH3 C SCoA
Propionyl-CoA Acetyl-CoA
Propionyl-CoA is converted to succinyl-CoA by propionyl-CoA carboxylase which has a
biotin prosthetic group.

- Succinyl-CoA can enter the citric acid cycle, but does not produce energy since succinyl-
CoA cannot metabolize to CO2 in the citric acid cycle. Note: Acetyl-CoA is the only
molecule that enters in the citric acid cycle and produce energy. (Do not misunderstand)

{Propionyl-CoA + <ATP>} → {Succinyl-CoA} → {Oxaloacetate + [GTP + FADH2 +

NADH]} → {Pyruvate + <GTP> + [ATP]} → {Acetyl-CoA + [NADH]} → {2CO2 +
[FADH2 + 3NADH + GTP] (in CAC)} = 2FADH2 + 5NADH + 2GTP - ATP = 4ATP +
15ATP + 2ATP – ATP = 20 ATP
- Succinyl-CoA is also converted to glucose in gluconeogenesis.
Succinyl-CoA →→ OAA → PEP →→ glucose

11
 Chapter 19 12 Takusagawa’s Note©

Example
#Acetyl- #Propionyl- #Oxidation 1st , 3rd = 2nd , 4th = CoA attach
CoA (12) CoA (20) (5) bond (-2) bond (-3) (-2)
C18Δ15 9 0 8 1 0 1
#ATP 108 0 40 -2 0 -2
Total 108 108 148 146 146 144

C18Δ12,15 9 0 8 1 1 1
#ATP 108 0 40 -2 -3 -2
Total 108 108 148 146 143 141

C18Δ9,12,15 9 0 8 2 1 1
#ATP 108 0 40 -4 -3 -2
Total 108 108 148 144 141 139

C18Δ6,9,12,15 9 0 8 2 2 1
#ATP 108 0 40 -4 -6 -2
Total 108 108 148 144 138 136

C18Δ6,9,12,15 9 0 8 2 2 0
-CoA
#ATP 108 0 40 -4 -6 0
Total 108 108 148 144 138 138

C19Δ6,9,12,15 8 1 8 2 2 1
#ATP 96 20 40 -4 -6 -2
Total 96 116 156 152 146 144

C19Δ9,12,15- 8 1 8 2 1 0
CoA
#ATP 96 20 40 -4 -3 0
Total 96 116 156 152 149 149

12
 Chapter 19 13 Takusagawa’s Note©

Methylmalonyl-CoA mutase contains a coenzyme B12, and catalyzes unusual carbon

skeleton rearrangement.
Methylmalonyl-CoA mutase contains Co atom in a hemelike corrin.
- One of six ligands is deoxyadenosyl group, i.e., C5⎯Co
SCoA SCoA
H C O O C H
H C C H H C C H
H CO2 H CO2
Methylmalonyl-CoA Succinyl-CoA

13
 Chapter 19 14 Takusagawa’s Note©

Mechanism of methylmalonyl-CoA mutase

1. C5⎯Co bond is cleaved when Ado-CH2 receives H from the substrate (H-C1).
2. The substrate form the C1⎯Co bond.
3. When the C1⎯Co bond is cleaved, rearrangement of X occurs, i.e., C1-X.
4. The H of Ado-CH3 is returned to C2 of the substrate.
5. Ado-CH2 forms a bond with Co, C5⎯Co, to regenerate the B12.

- This reaction begins with homolytic cleavage of C⎯Co bond (C••Co → C• + •Co).

14
 Chapter 19 15 Takusagawa’s Note©

Fatty acid oxidation also takes place in peroxisome

- Very long chain fatty acids are shortened by peroxisomal β-oxidation since the β-oxidation
of very long chain fatty acid is relatively slow in mitochondrion.
- The shorter chain fatty acids enter mitochondrial β-oxidation.
- Very long chain fatty acids diffuse into peroxisome. Thus carnitine is not utilized.
- Peroxisomal very long chain acyl-CoA synthetase is one of the unique enzyme.
- Deficiency of the peroxisomal acyl-CoA synthetase causes X-adenoleukodystrophy (X-
ALD).

Ketone Bodies

Acetoacetate
Acetyl-CoA +
β-hydroxybutyrate

Ketone Bodies
CAC

- Sometimes acetyl-CoA gets very high, then acetyl-CoA forms “Ketone bodies” in liver’s
mitochondria.
- Formation of ketone bodies from acetyl-CoA is shown in Fig. 23-22.

15
Chapter 19 16 Takusagawa’s Note©

β-hydroxybutyrate

16
 Chapter 19 17 Takusagawa’s Note©

- Ketone bodies are used as energy source by other tissues, heart and skeletal muscle.
- Although brain uses only glucose as its energy source, ketone bodies become the brain’s
major fuel source during starvation.
- Metabolism of ketone bodies is shown in Fig. 23-23.

- Accumulation of acetoacetate may cause non-enzymatic breakdown of acetoacetate to

acetone and CO2 as shown below. The resulting acetone is excreted by urea (ketoneurea)
and breath (sweet smell).

CO2
O O O
-
H3C C CH2 C O H3C C CH3
Acetoacetate Acetone

17
 Chapter 19 18 Takusagawa’s Note©

Fatty acid synthesis

Fatty acid synthesis takes place at liver (cytosol).
As always seen all biological processes, oxidation and synthesis take different pathways.
- In thermodynamics, if one direction is exergonic pathway, the opposite direction must be
endergonic pathway. Thus, it is impossible to occur both directions in the same system.
- It is easy to control the pathway if they are separated.
Differences between oxidation and synthesis are summarized in Fig. 23-25.

Fatty acid oxidation: takes place in mitochondrion and fatty acid carrier: CoA.
Fatty acid synthesis: takes place in cytosol and fatty acid carrier: acyl-carrier protein (ACP).
Both CoA and ACP have long phosphopantetheine groups in their structures.
The fatty acids are attached on the end of cysteamine residues (HS-CH2-CH2-NH-).

18
 Chapter 19 19 Takusagawa’s Note©

Acetyl-CoA carboxylase
- Acetyl-CoA carboxylase catalyzes the first committed step of fatty acid biosynthesis and one
of its rate-controlling steps.
- Malonyl-CoA is synthesized from acetyl-CoA and CO2 catalyzed by acetyl-CoA
carboxylase. In this reaction, biotin is the cofactor.
O ATP ADP + Pi O
HCO3- + CH3-C-SCoA -
OOC-CH2-C-SCoA
- Acetyl-CoA carboxylase is inhibited by palmitate (product) and activated by citrate.
- High [citrate] indicates high [acetyl-CoA], i.e., OAA + acetyl-CoA↑ ⎯→ citrate↑.
- Thus, the excess of acetyl-CoA enters into fatty acid synthesis.
- Malonyl-CoA carries an ATP hydrolysis energy which drives the condensation reaction to
produce acetoacetate-ACP.

Fatty acid synthase

Reaction sequence of palmitate synthesis is shown below.

19
 Chapter 19 20 Takusagawa’s Note©

- Fatty acid synthase is composed of multi-functional subunits. Those are:

β-ketoacyl-ACP synthase activity
malonyl-CoA-ACP transacetylase activity
acetyl-CoA-ACP transacetylase activity
β-hydroxyacyl-ACP dehydrase activity
enoyl-ACP reductase activity
β-ketoacyl-ACP reductase
acyl-carrier protein
palmitoyl thioesterase activity

- Fatty acid synthase is a dimer and has all enzyme activities.

- Multi-functional enzymes are very efficient since the substrate diffusion time is very short.

20
 Chapter 19 21 Takusagawa’s Note©

Start here

21
 Chapter 19 22 Takusagawa’s Note©

Sequential reactions of palmitate synthesis (shown in previous page)

1. Acetyl from acetyl-CoA binds on of Cys-SH of β-ketoacyl-ACP (β-keto acyl synthase).

2. Malonyl from malonyl-CoA binds on pant-SH of ACP.
3. Condensation reaction --- transfer acetyl group to malonyl group, and decarboxylation →
acetoacetyl-ACP and free the Cys-SH.
4. Reduction
5. Dehydration
6. Further reduction → butyryl-ACP.
7. Transfer of acyl group to Cys-SH (β-keto acyl synthase) and free pant-SH of ACP.
Repeat 2-7 steps six more times.
8. Transfer palmitoyl group to pant-SH and free Cys-SH.
9. Hydrolysis of C-S bond and release product (palmitate) and free pant-SH.

- All processes are carried out on the fatty acid synthase, and in cytosol.
- Overall reaction of palmitate synthesis is:
Acetyl-CoA + 7malonyl-CoA + 14NADPH + 7H+ →
palmitate + 7CO2 + 14NADP+ + 8CoA + 6H2O

Since 7malonyl-CoA are derived from acetyl-CoA as follows:

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

Therefore, overall stoichiometry is:

8Acetyl-CoA + 14NADPH + 7ATP → palmitate + 14NADP+ + 8CoA + 6H2O + 7ADP +7Pi

22
 Chapter 19 23 Takusagawa’s Note©

Transport of acetyl-CoA from mitochondrion to cytosol

Since acetyl-CoA is synthesized in a mitochondrion, acetyl-CoA must be transported to cytosol

via the tricarboxylate transport system.
1. Mitochondrial pyruvate is converted to oxaloacetate.
2. Acetly-CoA + oxaloacetate → citrate.
3. Citrate is transported to cytosol via tricarboxylate transport system (membrane protein).
4. Cytosol citrate is converted to oxaloacetate + acetyl-CoA
5. Oxaloacetate → malate → pyruvate in cytosol.

23
 Chapter 19 24 Takusagawa’s Note©

Elongation and desaturation (Post-modification)

- Elongation processes (elongases) are present in both the mitochondrion and endoplasmic
reticulum.
- In endoplasmic reticulum: the same processes of fatty acid biosynthesis (malonyl-CoA is
precursor).
- In mitochondrion: the reversed processes of β-oxidation, except for the final reaction that
uses NADPH rather than FADH2 as its redox coenzyme.

- Desaturation is catalyzed by the chain-length specific fatty acyl-CoA desaturases, such as Δ9-
fatty acyl-CoA desaturase.
- Mammalian cells contain: Δ4-, Δ5-, Δ6-, Δ9-fatty acyl-CoA desaturases.
- Plant cells contain : Δ4-, Δ5-, Δ6-, Δ9-, Δ12-, Δ15-fatty acyl-CoA desaturases.

24
 Chapter 19 25 Takusagawa’s Note©

- Double bond insertion occurs three carbons away from as existing on toward the CoA at the
carbon. For example,
C18Δ 9,12 -2H +2C -2H +2C
C18Δ 6, 9,12 C20Δ 8, 11,14 C20Δ 5, 8, 11,14 C22Δ 7, 10, 13, 16

- The desaturases are non-heme iron-containing enzymes that catalyze the general reaction:
H H O
+ +
CH3 (CH2)x C C (CH2)y C SCoA + NAD + H + O2
H H

O
CH3 (CH2)x C C (CH2)y C SCoA + 2H2O + NADH
H H
Actually, desaturase is complexed with other redox proteins, namely, desaturase, cytochrome b5,
and NADH-cytochrome b5 reductase as shown below.

Summary of β-oxidation and fatty acid biosynthesis

β-oxidation at mitochondrion:
1. By using 2 ATP, CoASH is attached to fatty acid to produce acyl-CoA.
2. Desaturation of Cβ-Cα bond by oxidation (cofactor: FAD → FADH2).
3. Hydration of -Cβ=Cα- to attach -OH to Cβ.
4. Oxidation of Cβ-OH to Cβ=O with cofactor (NAD+ → NADH).
5. Cleavage of Cβ-Cα bond by addition of CoASH.

Fatty acid biosynthesis at cytosol:

1. Formation of malonyl-CoA from acetyl-CoA + CO2
2. Reverse reactions of β-oxidation as described above. Note: different cofactors (both
reductions utilize NADPH).
3. Acyl group is grown on the end of acyl-carrier protein (ACP) instead of CoA in β-oxidation.

Synthesis of triacylglycerols
Triacylglycerols are synthesized from fatty acyl-CoA esters and glycerol-3-phosphate or
dihydroxyacetone phosphate (Fig. 23-34).

25
Chapter 19 26 Takusagawa’s Note©

26
 Chapter 19 27 Takusagawa’s Note©

Regulation of fatty acid metabolism

Major regulation site

27
 Chapter 19 28 Takusagawa’s Note©

Hormonal control
- Glucagon, epinephrine --- stimulate fatty acid oxidation.
Short effect: ↑cAMP, ↑adipose lipase
Long effect: ↓acetyl-CoA carboxylase (when the enzymes are phosphorylated, they are
connected to protomers. The protomers are inactive)
↓synthesis of acetyl-CoA carboxylation (synthesis of malonyl-CoA)
↓fatty acid synthesis
- Insulin --- stimulate fatty acid synthesis.
Short effect: ↓cAMP, ↑cAMP-independent protein kinase
Long effect: ↑acetyl-CoA carboxylase by dephosphorylation
↑synthesis of fatty acid
↓lipoprotein lipase
- Note: Effects of glucagon & epinephrine ↔ Insulin

Cholesterol metabolism
- Cholesterol is in cell membrane, and precursors of hormones, bile acids and vitamin D.
- Cholesterol has C27 skeleton and is produced from acetates that are first converted to
isoprene units, C5,
CH3
H2C C CH CH2
Isoprene

Acetate → isoprenoid intermediate (C5) → squalene (C30) → cyclization product (C30) →

cholesterol (C27).

Pathway of isoprenoid metabolism

28
 Chapter 19 29 Takusagawa’s Note©

Key enzyme

β-hydroxy-β-methylglutaryl-CoA (HMG-CoA) is a key cholesterol precursor

- HMG-CoA is synthesized from condensation of acetyl-CoA’s.
Thiolase
2(Acetyl − CoA) ⎯ ⎯⎯⎯ ⎯→ β − Ketobutyryl − CoA + Acetyl - CoA
HMG − CoA synthase
⎯ ⎯⎯⎯⎯⎯⎯⎯⎯ ⎯→ HMG − CoA
- Isopentenyl pyrophosphate and dimethylallyl pyrophosphate are synthesized from HMG-
CoA.
CH3 H2 CH3 H2
O O O O
C C - C C -
H2C C O P O P O H3C C O P O P O
H2 - - H - -
O O O O
Isopentenyl pyrophosphate Dimethylallyl pyrophosphate

29
 Chapter 19 30 Takusagawa’s Note©

2Acetyl-CoA

Thiolase

β -Ketobutyryl-CoA + CoA

Acetyl-CoA HGM-CoA synthase

β -Hdroxy-β -methylglutaryl-CoA (HMG-CoA)

HMG-CoA reductase HMG-CoA lyase

(cytosol) (mitochondria)

Mevalonate β -Ketobutyrate
+ +
CoA Acetyl-CoA

Keytone bodies

30
 Chapter 19 31 Takusagawa’s Note©

- The last step of reaction is an ATP-dependent concerted dehydration-decarboxylation.

- Formation of dimethylallyl pyrophosphate reaction is catalyzed by isopentenyl

pyrophosphate isomerase.

31
 Chapter 19 32 Takusagawa’s Note©

- Formation of squalene is:

- 2 times head-tail condensation reactions to produce geranyl pyrophosphate (C10 unit) →
farnesyl pyrophosphate (C15 unit).
- head-head condensation of two resulting C15 units (farnesyl pyrophosphate) to produce
squalene.

32
 Chapter 19 33 Takusagawa’s Note©

- Cyclization of squalene is shown below (epoxization is an essential process).

- C30 lanosterol is the product of cyclization of squalene.

- Additional 19 step reactions are required to convert lanosterol to cholesterol.

33
 Chapter 19 34 Takusagawa’s Note©

Summary of cholesterol biosynthesis

1. HMG-CoA (C6 fragment) is synthesized from three acetyl-CoA.
2. HMG-CoA is converted to mevalonate, and then two isoprene pyrophosphates (isopentenyl-
PPi and dimethylallyl-PPi).
3. Isopentenyl-PPi and dimethylallyl-PPi are connected by head-tail to geranyl-PPi.
4. Another isopentenyl-PPi and geranyl-PPi is connected by head-tail to farnesyl-PPi.
5. Two farnesyl-PPi are put head-head condensation to produce the C30 fragment, squalene.
6. Squalene is cyclized to form lanosterol.
7. Lanosterol is converted to C27 cholesterol by 19 step reactions.

- From cholesterol:
Cholesterol
Bile acid (cholic acid)
glucocorticoids
(cortisol)
Androgens Progestins (progesterone)
(testesterone)

Mineralcorticoids (aldosterone)
Estrogens
(estradiol)

Cholesterol synthesized by the liver is:

1. converted to bile acids for use in digestive process.
2. esterified to form cholesterol ester and secreted into bloodstream as part of the very low
density lipoprotein (VLDL)

O
R C O

34
 Chapter 19 35 Takusagawa’s Note©

Cholesterol Transport

Synthesis (in liver)

Phosphatidylcholine (PC) Bile acids (in gallbladder)

LCAT

Cholesterol ester VLDL Secreted into bloodstream

LCAT = Lecithin-cholesterol acyl transferase

Formation and secretion of VLDL in liver is illustrated below.

Review of Fig. 11-53 for VLDL, IDL, LDL and

HDL.

- VLDL/IDL/LDL delivers cholesterol to

tissues.
- HDL delivers cholesterol to liver.
- LDL is the major cholesterol carrier of the
bloodstream.

Fig. 11-50.

35
 Chapter 19 36 Takusagawa’s Note©

LDL enters into cells through endocytosis. LDL is recognized by the LDL receptors which are
synthesized on the endoplasmic reticulum.

ACAT = Acyl-CoA Cholesterol Acyltransferase

Note:
- Cholesterols released from LDL in cells enters into endoplasmic reticulum and are utilized to
form membrane of ER.
- Presence of cholesterol in cytosol decreases the rate of:
HMG-CoA reductase synthesis.
LDL receptors synthesis.

36
 Chapter 19 37 Takusagawa’s Note©

Control of cholesterol levels

HMG-CoA reductase is the primary control site for cholesterol biosynthesis.

2NADH 2NAD+ CoA

HO CH3 HO CH3
C C
H2C CH2 C SCoA H2C CH2 CH2 OH
C O HMG-CoA C
-O O reductase -O O
Mevalonate
HMG-CoA

1. Hormonal regulation of HMG-CoA reductase

- HMG-CoA reductase activity is regulated by phosphorylation/dephosphorylation with
protein kinases (PKs) and phosphatases.

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 Chapter 19 38 Takusagawa’s Note©

- This enzyme cascade system is quite similar to the glycogen metabolism regulation.
- When [cAMP] ↑, activity of HMG-CoA reductase ↓ and thus cholesterol biosynthesis ↓.
- The [cAMP] is regulated by hormones, i.e., insulin and glucagon.
Insulin: ↓ [cAMP], thus ↑ cholesterol biosynthesis (in general, ↑ all biosyntheses).
Glucagon: ↑ [cAMP], thus ↓ cholesterol biosynthesis (in general, ↓ all biosyntheses).

- Compactin and Lovastatin (fungal products) are competitive inhibitor of HMG-CoA

reductase. Lovastatin is now in routine use for treatment of hypercholesterolemia.
HO -
COO
R = H Compactin
OH R = CH3 Lovastatin
O

O
CH3 CH3

2. Regulation of synthesis of HMG-CoA reductase mRNA

- Presence of relatively high cholesterol in cytosol decreases the synthesis of mRNA of HMG-
CoA reductase.

3. Degradation of HMG-CoA reductase

- The half-life (t1/2) of HMG-CoA reductase is decreased when [cholesterol] is increased.
- Cholesterol in cytosol stimulates degradation (proteolysis: cut peptide bonds) of the C-
terminal domain of HMG-CoA reductase, which is the catalytic domain.

Proteolysis

The other regulation of cholesterol levels are:

- LDL receptor synthesis (is decreased by high [cholesterol] in cytosol).
- Regulation of LCAT (lecithin-cholesterol acyl transferase).

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 Chapter 19 39 Takusagawa’s Note©

LDL level in blood depends on the rate of removal from bloodstream by liver.
(a) If liver has enough LDL receptor, the
[cholesterol] in blood is decreased.

(b) Genetically defective LDL receptor

synthesis cannot produce enough
LDL receptor, thus cholesterol levels
in blood of these individuals are
relatively high and called Familial
hypercholesterolemia (FH).

(c) High cholesterol diet decrease the

LDL receptor synthesis, since the
dietary cholesterol enters liver and
the [cholesterol] is increased. High
[cholesterol] in liver decreases the
LDL receptor synthesis.

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 Chapter 19 40 Takusagawa’s Note©

Complexed lipids
Typical complex lipids are: glycerolipid and sphingolipid
O OH H
O CH2 O C R1 O CH C C (CH2)12 CH3
R2 C O C H R2 C NH C H H
CH2 O X CH2 O X

1,2-diacyl-syn-glycerol N-acylsphingosine (ceramide) X=H

Glyceroglycolipid Sphingoglycolipid X = carbohydrate
Glycerophospholipid Sphingophospholipid X = phosphate

Glycerophospholipid syntheses
Phosphatidylcholine and
phosphoethanolamine syntheses
- Polar head groups (phosphatidylcholine
and phosphoethanolamine) are activated
by attaching CDP.
- Note: CTP is used for energy source.
UTP is used for glycogen synthesis.
(Carbohydrates).

- Activated CDP-choline, CDP-

ethanoamine, etc. are attached to
diacylglycerol as shown below.

Fig. 23-67

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 Chapter 19 41 Takusagawa’s Note©

Phosphatidylserine synthesis
- Ethanoamine group of phosphatidylethanolamine is replaced with serine by head group
exchange reaction.

Cardiolipin synthesis
- Cardiolipin is synthesized by two phosphatidylglycerol condensation reaction.
- Cardiolipin is an important phospholipid in heart tissue.

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 Chapter 19 42 Takusagawa’s Note©

Phosphatidylglycerol and phosphatidylinositol syntheses

- Diacylglycerol portion (phosphatidic acid) is activated by attaching CMP (CDP-
diacylglycerol)
- Then glycerol-3-phosphate and inositol are connected by using the CMP hydrolysis energy.
- Phosphatidylinositol is an important signal transduction substance (second messenger).

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 Chapter 19 43 Takusagawa’s Note©

Sphingolipids biosynthesis
- Basic fragments of sphingosine and ceramide (N-acetylsphingosine) are synthesized from
palmitoyl-CoA, serine and acyl-CoA.

Palmitate

Acyl
group

Serine
43
 Chapter 19 44 Takusagawa’s Note©

- Sphingomyelin is synthesized from ceramide (N-acetylsphingosine) and

phosphatidylcholine.

- Cerebrosides (ceramide + sugar) is

synthesized from ceramide and UDP-sugar
(such as UDP-glucose, UDP-galactose).

44