SBT 401 GENERAL BIOCHEMISTRY

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LIPID METABOLISM

Lawrence Nugbienyo (PhD.) FACULTY OF APPLIED SCIENCES

lnugbienyo@atu.edu.gh
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Introduction
 Triacylglycerols – energy reserve and body
Synthesis of
lipoproteins
Synthesis of
chylomicrons from insulator.
(VLDL) dietary lipids

Liver
Small intestine  Phospholipids, glycolipids and cholesterol –

VLDL with TG
Chylomicrons
with
major components of cell membranes.
transported TG transported
Stored
triacylglycerol
 Arachidonic acid: synthesis of intercellular
Adipose
tissue
regulators – prostaglandins, thromboxanes,
Fatty
acids
prostacyclins, etc.
Triacylglycerols  Transport lipids: chylomicrons, lipoproteins
and fatty acids utilized
ENERGY
Muscle, liver, heart etc. (VLDL, LDL, HDL)
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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
Lipolysis
O
O CH2 C O R
1 CH2 OH R1COOH
Lipolysis
R2 C O CH HO CH + R2COOH
+ 3H2O
CH2 C O R
3 CH2OH R3COOH
O
Triacylglycerol Glycerol Free fatty acids (3)

Lipolysis is the complete degradation of triacylglycerol to form diacylglycerol. The

triacylglycerol to glycerol and free fatty other two fatty acids are cleaved by
acids. Triacylglycerol lipase removes the additional lipases specific for
fatty acid either from carbon 1 or 3 of the diacylglycerol and monoacylglycerol.
and are transported in a bound form t
SBT 401 GENERAL BIOCHEMISTRY
3
The
FACULTY OF APPLIED SCIENCES
free
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY fatty acids enter various tissu
_____________
Lipid Metabolism
Regulation
288
of TG Lipase BIOCHEMISTRY

ATP
Epinephrine
Norepinephrine
Glucagon, Thyroxine Adenylate cyclase
Glucocorticoids Hormone sensitive
TSH, ACTH, GH TG lipase b
(inactive)

Insulin, niacin, PGE1 ATP

Pi

cAMP Protein Phosphatase

kinase
Insulin
Phosphodiesterase
Caffeine ADP
Hormone sensitive
5 ’ AMP TG lipase a
(active)

Triacylglycerol Diacylglycerol

Free fatty acid FFA

Monoacylglycerol

FFA
Glycerol

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PGE 1OF
– Prostaglandin E1; TG–Triacylglycerol; FFA–Free fatty acid). _____________
DEPARTMENT SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
 CH2 OH III. � -Oxidation proper
HO CH Fate of Glycerol
Each cycle of � -oxidation, liberating a two
CH2 OH carbon unit-acetyl CoA, occurs in a sequence of
Glycerol four reactionsFig.14.7
( ).
1. Oxidation : Acyl CoA undergoes
ATP
Glycerokinase
❖ Glycerol is phosphorylated in the liver.
dehydrogenation by an FAD-dependent
ADP
flavoenzyme, acyl CoA dehydrogenase.A
double bond is formed between � and � carbons
(i.e., 2 and 3 carbons).

HO CH
CH2 OH ❖ Glycerol 3-phosphate may be used for the synthesis of
2. Hydration : Enoyl CoA hydratase brings
about the hydration of the double bond to form
CH2 O P
Glycerol 3-phosphate triacylglycerols and phospholipids.
� -hydroxyacyl CoA.
3. Oxidation : � -Hydroxyacyl CoA dehydro-
genase catalyses the second oxidation and gene-
G d
lyc eh

rates NADH. The product formed is � -ketoacyl

er y d

Glycerol 3-phosphate may also enter glycolysis by

ol ro

CoA. ❖
3- ge

+
ph n

NAD
4. Cleavage : The final reaction in
os ase
ph

+
NADH + H � -oxidation
is the liberation of a 2 carbon
at

getting converted to dihydroxyacetone phosphate.

e

fragment, acetyl CoA from acyl CoA. This occurs

by a thiolytic cleavage catalysed by
�-ketoacyl
CoA thiolase(or simply thiolase).
CH2 OH
The new acyl CoA, containing two carbons
C O
less than the original, reenters the � -oxidation
CH2 O P cycle. The process continues till the fatty acid is
Synthesis of
triacylglycerols,
Dihydroxyacetone completely oxidized.
phosphate
phospholipids

Glycolysis R CH2 CH2 COO–

Fatty acid
Fig. 14.4 : Fate of glycerol.
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APPLIED SCIENCES
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DEPARTMENT OF SCIENCE LABORATORY
Thiokinase PyrophosphataseTECHNOLOGY
It should be noted that the coenzyme A used PPi 2Pi Lipid Metabolism
Fate of Free Fatty Acids

❖ Triacylglycerol / Fatty Acid Cycle: About 65% of FFA are converted to TG, and
sent back to adipose tissue for deposition.
❖ About 95% of the energy obtained from fat comes from the oxidation of fatty
acids.
❖ Fatty acids are mostly oxidized by β – oxidation: the oxidation of fatty acids on
the β – carbon atom, resulting in the sequential removal of acetyl CoA.

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
β – oxidation of Fatty Acids

Three Stages:

1. Activation of fatty acids.

2. Transport of fatty acids into mitochondria.

3. β - oxidation proper in the mitochondrial matrix.

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
less than the original, reenters the � -oxidation
cycle. The process continues till the fatty acid is
β – oxidation (Fatty Acid Activation)
completely oxidized.

R CH2 CH2 COO– 

Fatty acids are activated to acyl CoA by
Fatty acid
thiokinases or acyl CoA synthetases.
ATP
Thiokinase Pyrophosphatase
PPi 2Pi

Two high energy phosphates are utilized.
O
R CH2 CH2 C AMP 
Pyrophosphatase hydrolyses PPi to
Acyladenylate
phosphates (Pi).
CoASH

AMP

O
R CH2 CH2 C ~ CoA
Acyl CoA

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
 β 290
– oxidation (Transport of Acyl CoA into Mitochondria)
BIOCHEMISTRY

Cytosol Inner Mitochondrial

mitochondrial matrix
membrane

O O
R C SCoA Carnitine Carnitine R C SCoA
Acyl CoA Acyl CoA

Carnitine acyl- Carrier Carnitine acyl-

transferase I protein transferase II

O O
CoASH R C Carnitine R C Carnitine CoASH
Acyl-carnitine Acyl-carnitine

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9 FACULTY
Fig. 14.6 : Carnitine OF APPLIED
shuttle SCIENCESof activated fatty acid (acyl CoA) into mitochondria.
for transport _____________
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
 O 8 Acetyl CoA (2C)

β – oxidation (Proper)
β α
R CH2 CH2 CH2 C SCoA TCA 96 ATP (8�12)
Acyl CoA cycle

FAD
2ATP Acyl CoA Fig. 14.8 : An overview of oxidation of palmitic acid.
(1) dehydrogenase
ETC FAD H2
O
Occurs in a sequence of four reactions:
SIDS—a disorder due
R CH2 CH CH C SCoA to blockade in � - oxidation
2 trans
-enoyl CoA
Δ
The sudden

Oxidation: Acyl CoA dehydrogenation.
infant
death syndrome
an unexpected death of
(SIDS)is
healthy infants,
usually
H 2O
Enoyl CoA
(2) hydratase overnight. The real cause of SIDS is not known.
It is now estimated that at least 10% of SIDS is
OH O

Hydration: Formation of β-hydroxyacyl CoA.
due to deficiency of medium chain acyl CoA
R CH2 CH CH2 C SCoA dehydrogenase. The enzyme defect has a
β -Hydroxyacyl CoA
frequency of1 in 10,000 births and is,in fact,

3ATP NAD
+ more prevalent

Oxidation: Formation of β-ketoacyl CoA.
than phenylketonuria.The
occurrence of SIDS is explained as follows
(3) β-Hydroxyacyl CoA
dehydrogenase
ETC NAD H + H
+
Glucose is the principal source of energy,
O O Cleavage: Liberation of acetyl CoA.
soon after eating or feeding babies. After a few
hours, the glucose level and its utilization
R CH2 C CH2 C SCoA decrease and the rate of fatty acid oxidation must
β -Ketoacyl CoA
simultaneouslyincrease to meet the energy
CoASH
Krebs needs. The sudden death in infants is due to a
Thiolase cycle caused by a defici-
(4) 2CO2 blockadein �-oxidation
O O ency in medium chain acyl CoA dehydrogenase
R CH2 C SCoA +CH3 C SCoA
(MCAD).
Disorders: SIDS, JVS
Acyl CoA (–2C) Acetyl CoA
Jamaican vomiting sickness

Fig. 14.7 : �
-Oxidation of fatty acids : Palmitoyl CoA This disease is characterizedby severe
hypoglycemia, vomiting, convulsions, coma and SBT 401 GENERAL BIOCHEMISTRY
10carbon) undergoes seven cycles to yield 8 acetyl
(16 FACULTY OF APPLIED SCIENCES
_____________
CoA [I–Activation; II–Transport; III– �Oxidation
DEPARTMENT
proper— death. It is caused
OF SCIENCE by eating
LABORATORY unripe ackee fruit
TECHNOLOGY
Lipid Metabolism
(1) Oxidation, (2) Hydration, (3) Oxidation and
 Oxidation of Odd Carbon Chain Fatty Acids
BIOCHEMISTRY

rogenase and thus

� CO 2
CH3 Propionyl CoA CH3
ns. carboxylase
CH2 H C COO–
Biotin
of odd carbon CO S CoA ATP ADP + Pi
CO S CoA
acids Propionyl CoA D-Methylmalonyl CoA

oxidation of Methyl-
odd numberof malonyl CoA
COO– racemase

CH2 CH3
Methylmalonyl CoA
s that in the last CH2 mutase, B12
–OOC C H
CO S CoA CO S CoA
Succinyl CoA L-Methylmalonyl CoA
B12 deficiency
CoA which is
A as follows (Fig.14.9
) TCA cycle Methylmalonic acid

SBT 401 GENERAL BIOCHEMISTRY

of ATP, CO
2 and vitamin
11 FACULTY OF APPLIED SCIENCES
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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
ylmalonyl CoA. Lipid Metabolism
α-oxidation and ω-oxidation

 Removal of one-carbon unit at a time.

 Refsum’s Disease: deficiency of phytanic acid α-oxidase.

 ω-oxidation involves hydroxylation followed by oxidation of ω-carbon

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
 O O

ate
COO– CH3 C S CoA + 3CHC S CoA
Acetyl CoA Acetyl CoA Ketogenesis & Utilization of Ketone
296
Bodies
CoA SH β Ketothiolase
OH pr
CH3 CH CH2 COO– ha
odies.
CH3
O
C CH
O
2 C S CoA
 Synthesis of ketone bodies in the β -Hydroxybutyrate cy
ac
Acetoacetyl CoA +
O NAD sy
CH3 C S CoA
HMG CoA synthase
liver. +
β-Hydroxybutyrate
dehydrogenase
(o
re
NADH + H
CoA SH di
Acetyl CoA,
atrix O
–
OOC CH
2 C CH
2 C S CoA
OH O  Ketogenesis regulation: HMG CH3 C CH2 COO–
ap
Acetoacetate
CH3 in

condense to
β-Hydroxy-
β-methylglutaryl CoA (HMG CoA)

O
CoA synthase, glucagon, insulin. CH2 COO–
CH2 CO SCoA
pr
en
CH3 C S CoA HMG CoA lyase Succinyl CoA Thiophorase ca
Acetyl CoA ur
–
OOC CH
2 C
O  In diabetes mellitus, starvation. CH2 COO–
CH2 COO–
n. CH3 Succinate as
Acetoacetate O O im
 Ketosis: ketonemia, ketonuria. CH3 C CH2 C SCoA in
β-Hdeh
us

CoA). le
eo

yd yd

Acetoacetyl CoA
n

ro rog
ta

on, its
xy e
on

+
bu na

NADH + H
Sp

s
ty s e

CO2
CoASH di
ra

Ketoacidosis
te

+ Thiolase
HMG CoA to NAD  pl
tyl CoA. ex
O O O
OH
spontaneous
CH3 C CH – CH3 C SCoA + CH
3 C SCoA
ne. 3 OOC CH
2 C CH
3 Re
Acetone
H Acetyl CoA Acetyl CoA
uced by a
β-Hydroxybutyrate
ybutyrate.
ov
me amino acids SBT 401 GENERAL BIOCHEMISTRY
13 FACULTY OF APPLIED SCIENCES av
acetate or acetyl _____________
bodies, e.g.
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
is
sources of energy for the such
peripheral tissues ac
Summary of Synthesis, Utilization and Excretion of
Chapter 14 : METABOLISM OF LIPIDS 297
Ketone Bodies
EXTRAHEPATIC TISSUES
LIVER BLOOD (e.g. muscle)

Fatty acids

Ketone bodies

Kidneys Lungs
Fatty acids Fatty acids
Glucose Amino Amino
acids Glucose acids
Acetyl CoA
Ketone Ketone
bodies bodies Acetyl CoA
TCA
cycle

TCA
2CO 2 cycle
Ketone bodies Acetone
(in urine) (exhaled) 2CO 2

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
Ketoacidosis for fatty acid production is located in
machinery
Biosynthesis of Fatty Acids
❖ De novo synthesis of fatty acids occurs ❖ Fatty acid synthesis is in 3 stages:
predominantly in liver, kidney, adipose ●
Production of acetyl CoA and NADPH.
tissue and lactating mammary glands.
●
Conversion of acetyl CoA to malonyl
❖ Acetyl CoA is the source of carbon atoms.
CoA.
❖ NADPH provides the reducing equivalents. ●
Reactions of fatty acid synthase
❖ ATP supplies energy for fatty acid complex.
formation.

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
Production of Acetyl CoA and NADPH
Glucose Cytosol Mitochondrial
matrix

Pyruvate Pyruvate

HMP shunt NADPH + H+

CO2 Malic
enzyme
NADP+ Pyruvate
dehydrogenase
Malate Pyruvate
carboxylase
NAD+
Malate Amino Fatty
dehydrogenase acids acids
Fatty NADH + H+
acid Oxaloacetate
Acetyl CoA Oxaloacetate
ADP + Pi
Acetyl CoA Citrate Citrate
ATP lyase synthase
CoASH
Citrate Citrate

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
Formation of Malonyl CoA

O
CH3 C SCoA
Acetyl CoA
CO2
Acetyl CoA
ATP carboxylase
ADP + Pi Biotin
O
–
OOC CH
2 C SCoA
Malonyl CoA

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
 O β -Hydroxyacyl-ACP
CH3 C SCoA (5)
β -Hydroxyacyl-ACP
Acetyl CoA H 2O dehydratase
Cys SCoA
(1) Cys SH
ACP SH O
Acetyl CoA
Fatty acid CoASH ACP ACP S C CH CH CH
3
synthase transacylase
Trans Δ2 -Enoyl ACP
Cys SH O + (6)
NADP H + H
ACP S C CH3 Enoyl ACP
NADP
+ reductase
Acetyl-S-ACP Transfer of acetyl
to cysteine
Cys SH
O O O

Extra-Mitochondrial
– ACP S C CH2 CH2 CH3
Cys S C CH3 OOC CH2 C SCoA
Malonyl CoA Acyl-ACP (butyryl-ACP)
ACP SH

Synthesis of Fatty Acids

(2) Transfer of carbon
Acetyl S-enzyme Malonyl CoA-ACP chain from ACP to Cys
CoASH transacylase
O
O
Cys S C CH2 CH2 CH3
Cys S C CH3
O
ACP SH
ACP S C CH2 COO–
Acyl-S-enzyme
Acylmalonyl enzyme
(3)
β-Ketoacyl-ACP
CO2 synthase Reactions 2-6
repeated six
more times
Cys SH
O O Cys SH
O
ACP S C CH2 C CH3
β -Ketoacyl-ACP ACP S C (CH2)13 CH2 CH3
+
NADP H + H
(4) β -Ketoacyl-ACP H 2O Palmitoyl
SBT 401 GENERAL BIOCHEMISTRY + reductase thioesterase
_____________ NADP
Lipid Metabolism Cys SH
O OH Cys SH
+ CH3 CH2 (CH2)13 COO–
ACP S C CH2 CH CH3 ACP SH
18 FACULTY OF APPLIED SCIENCES β-Hydroxyacyl-ACP PALMITATE
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
contd. next column
Desaturation & Elongation of Fatty Acids

❖ Fatty acyl CoA desaturase catalyses the
formation of unsaturated fatty acids.
❖ Desaturation also involves
cytochrome b5 reductase, NADH and O2 .
❖ Essential fatty acids:
●
Linoleic acid (18:2 Δ9, 12)
●
Linolenic acid (18:3Δ9, 12, 15)
❖ Chain elongation takes place in mitochondria or
in endoplasmic reticulum (microsomes).
❖ Microsomal chain elongation is more
predominant: involves successive additions
flavin-dependent of malonyl CoA with the participation of
elongases and NADPH.
❖ Mitochondrial chain elongation is almost a
reversal of β-oxidation of fatty acids: Acetyl
CoA molecules are successively added to
fatty acid to lengthen the chain. The reducing
equivalents are derived from NADPH.

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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
Lipid Metabolism
Comparison component Fatty acid synthesis β-oxidation
Major tissues Liver, adipose tissue Muscle, liver
Subcellular site Cytosol Mitochondria
Precursor/substrate Acetyl CoA Acyl CoA
End product Palmitate Acetyl CoA
Intermediates are bound to Acyl carrier protein Coenzyme A
Coenzyme requirement NADPH (reducing equivalents) FAD and NAD+
Carbon units added/degraded Malonyl CoA Acetyl CoA
Transport system Citrate: mitochondria to cytosol Carnitine: cytosol to mitochondria
Inhibitor Long chain acyl CoA (inhibits Malonyl CoA (inhibits
acetyl CoA carboxylase) carnitine acyltransferase I)
The pathway increased After rich carbohydrate diet In starvation
Hormonal status that promotes High ratio of insulin/glucagon Low ratio of insulin/glucagon
Status of enzyme(s) Multifunctional enzyme complex Individual enzymes
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DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY Lipid Metabolism
 CH2 OH Glucose
HO C H
CH2 OH
Glycerol Glycolysis
ATP
Glycerol
kinase
ADP

CH2 OH Glycerol 3-phosphate CH2 OH

dehydrogenase
HO C H C O
CH2 O P CH2 O P
Glycerol 3-phosphate NAD+ NADH + H+ Dihydroxyacetone
O phosphate (DHAP)
O
R1 C SCoA
Glycerol 3-phosphate R1C~SCoA
acyltransferase DHAP acyl-

Biosynthesis of
CoASH transferase
CoASH
O O
CH2 O C R NADP+ NADPH + H+ CH2 O C R

Phospholipids
1 1
HO C H C O
CH2 O P 1-Acyl DHAP reductase CH2 O P
Lysophosphatidic acid 1-Acyl DHAP
O
R2 C SCoA
Acyltransferase
CoASH

O O
O CH2 O C R
1 O CH2 O C R
1
Phosphatase
R2 C O C H R2 C O C H
CH2 O P CH2 OH
H 2O Pi
Phosphatidic acid Diacylglycerol

O
R3 C SCoA
SBT 401 GENERAL BIOCHEMISTRY Acyltransferase
_____________ PHOSPHOLIPIDS CoASH
Lipid Metabolism
O
O CH2 O C R
1

21 FACULTY OF APPLIED SCIENCES R2 C O C H O

DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY CH2 O C R
3
Triacylglycerol
 O
O CH 2 O C R1
R2 C O C H
CH 2 O P
Phosphatidic acid
Choline
(Ethanolamine) H 2O CTP
ATP
Pi PPi
(1) (3) (5)
ADP

Phospho choline O O
Biosynthesis of (Phospho ethanolamine )
CTP
R2
O
C O C H
CH 2 O C R1
R2
O
C O C H
CH 2 O C R1

Phospholipids (cont.)
(2)
CH 2 OH CH 2 O P P
PPi
1,2-Diacylglycerol
CDP -choline Cytidine
(CDP -ethanolamine ) CDP-diacylglycerol
Glycerol
CMP (4) Inositol 3-phosphate

CMP CMP
(6)
O
O CH2 O C R1 O
O CH2 O C R1 Phosphatidylglycerol
SBT 401 GENERAL BIOCHEMISTRY R2 C O C H 3-phosphate
_____________ CH2 O P R2 C O C H H2O
Lipid Metabolism CH2 O P P
Choline Pi
( Ethanolamine )
Inositol Phosphatidylglycerol
Phosphatidyl choline Phosphatidyl inositol Phosphati-
(Phosphatidyl ethanolamine) dylglycerol
Phosphatidyl- Phosphatidyl
ethanolamine choline Glycerol
Biosynthesis of phospholipids [The enzymes are numbered— (1) Choline kinase, (2) Phosphocholine
Serine Cardiolipin
cytidyltransferase, (3) Phosphatidate phosphohydrolase, (4) Phosphocholine diacylglycerol transferase, (5) CTP–
CO2 (Diphosphatidylglycerol)
Phosphatidate cytidyltransferase, (6) CDP–Diacylglycerol inositol transferase].

Ethanolamine
22 FACULTY OF APPLIED SCIENCES
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY Phosphatidyl-
serine
 THANK YOU

SBT 401 GENERAL BIOCHEMISTRY

FACULTY OF APPLIED SCIENCES _____________
DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY Lipid Metabolism