Chapter 22

Lipid Biosynthesis

Biosynthesis of fatty
acid
 1. Fatty acid synthesis takes a different
pathway from its degradation
Occurs in the cytosol
Acetyl-CoA provides the first two carbons, which is elongated
by sequential addition of two-carbon units donated from
malonyl-CoA.
Intermediates are attached to the -SH groups of an acyl carrier
protein (ACP).
NADPH is the reductant.
The enzymes are associated as a multi-enzyme complex or
even being in one polypeptide chain in higher organisms (fatty
acid synthase).
Elongation by the fatty acid synthase
complex stops upon formation of palmitate
(C16), further elongation and desaturation are
carried out by other enzyme systems.
 2. Malonyl-CoA is formed from acetyl-
CoA and bicarbonate
HCO3- is required for fatty acid synthesis.
Acetyl-CoA carboxylase catalyzes this
carboxylation reaction.
The enzyme has three functional parts: a biotin
carrier protein; an ATP-dependent biotin
carboxylase; and a transcarboxylase.
.
biotin Trans-
carboxylase carboxylase

Acetyl-CoA carboxylase
catalyzes the two-step
carboxylation reaction
of acetyl-CoA in two
active sites.
3. The acetyl and malony groups are first
transferred to two –SH groups of the
fatty acid synthase complex
 The acetyl group of acetyl-CoA is first transferred to the –
SH group of a Cys residue on the β -ketoacyl-ACP synthase
(KS) in a reaction catalyzed by acetyl-CoA-ACP
transacetylase (AT).
 The malonyl group is transferred from malonyl-CoA to the –
SH group of the 4`-phosphopantetheine covalently attached
to a Ser residue of the acyl carrier protein (ACP).
 4. Fatty acids are synthesized by a
repeating four-step reaction sequence
In the condensation reaction (step 1), catalyzed by β -ketoacyl-
ACP synthase, the methylene group of malonyl-CoA (linked to
ACP) undergoes a nucleophilic attack on the carbonyl carbon of
the acetyl group linked to KS, forming the β -ketobutyryl-ACP
with simultaneous elimination of CO2.
the β -ketobutyryl-ACP is then reduced to D-β -hydroxybutyryl-
ACP (step 2), using NADPH and the β -ketobutyryl-ACP
reductase (KR).
A water molecule is then removed from the β -hydroxybutyryl-ACP to
produce trans-∆ 2-butenoyl-ACP in a reaction catalyzed by β -
hydroxybutyryl-ACP dehydratase (step 3).
 A further reduction (step 4), also using NADPH, of the carbon-carbon
double in trans-∆ 2-butenoyl-ACP, catalyzed by enoyl-ACP reductase
produces a saturated acyl on ACP (butyryl-ACP).
The butyryl group is then transferred to the Cys –SH group of β -
ketoacyl-ACP synthase for another round of four reactions, which will
extend the chain by two more carbons.
Seven rounds of the four-step lengthening reactions
produces palmitoyl-ACP, which will be hydrolyzed to
release a free palmitate.

A total of 7 ATP and 14 NADPH will be consumed for

making one palmitate molecule.
6. Fatty acid synthesis occurs in cellular
compartments having a high
NADPH/NADP+ ratio
NAD and NADP have selected for functioning
as electron carriers in oxidative catablism and
reductive anabolism respectively.
In the hepatocytes and adipocytes, NADPH is
mainly produced in the cytosol via the pentose
phosphate pathway and by the malic enzyme.
.
 Pentose phosphate
pathway

Malic
enzyme

NADPH in the cytosol of animal cells

is largely produced by the oxidative
decarboxylation of malate and the
pentose phosphate pathway
 7. The acetyl groups of the
mitochondrion are transported into the
cytosol in the form of citrate
The acetyl-CoA molecules are made from glucose and
amino acids in mitochondria.
The are shuttled into the cytosol in the form of citrate
via the citrate transporter of the inner membrane.
Acetyl-CoA is regenerated by the action of ATP-
dependent citrate lyase in the cytosol.
Oxaloacetate is shuttled back into the mitochondria as
malate or pyruvate.
 8. The rate of fatty acid biosynthesis is
controlled by acetyl-CoA carboxylase
Excess fuel is generally converted to fatty
acids/triacylglycerol for longer term storage.
Acetyl-CoA carboxylase, catalyzing the committing and
rate-limiting step of fatty acid synthesis, is allosterically
inhibited by palmitoyl-CoA and activated by citrate.
Glucagon and epinephrine triggers the phosphorylation and
disassociation of the polymeric enzyme subunits, which
inactivates the enzyme.
Citrate partially activate the phosphorylated
acetyl-CoA carboxylase (similar to how AMP
partially active the dephosphorylated glycogen
phosphorylase).
 Dephosphorylated
acetyl-CoA
Carboxylase
(active)

Acetyl-CoA carboxylase
is regulated by allosteric
effectors and reversible
phosphorylation
 Citrate partially activate the
phosphorylated acetyl-CoA carboxylase
 11. Newly synthesized fatty acids have
mainly two alternative fates in cells
Fate I: be incorporated into tri-acylglycerol as
a form to store metabolic energy in long terms.
Fate II: be incorporated into membrane
phospholipids (during rapid growth).
 12. Phosphatidic acid is the common
precursor for the syntheses of both tri-
acylglycerol and glycerol-
phospholipids
 Phosphatidic acid (or diacylglycerol 3-phosphate) is made
by transferring two acyl groups from two acyl-CoAs to L-
glycerol 3-phosphate, which is derived from either glycerol
or dihydroxyacetone phosphate.
 A phosphatidic acid is converted to a triacylglycerol via a
dephosphorylation reaction (catalyzed by phosphatidic acid
phosphatase) and a acyl transferring reaction.
Phosphatidic
acid is derived
from L-glycerol 3-
phosphate and two
acyl-CoAs.

Often saturated
Often unsaturated
phospholipidic acid
is the common
precursor for both
triacylglycerols Phosphatidic acid
and phosphatase

glycerophospholi
pids
 Summary
Fatty acid biosynthesis takes a different pathway from
the reverse of its degradation and takes place in
different cellular compartments.
The aceytl-CoA units are transported out of
mitochondrial matrix as citrate.
Acetyl-CoA carboxylase catalyzes the rate-limiting
step of fatty acid synthesis and is highly regulated by
allosteric and covalent modifications.
 Palmitate, the usual final product of fatty acid synthesis, can be
further elongated and desaturated in sER.
.
 Phosphatidic acid (diacylglycerol 3-phosphate) is the common
precursor of both triacylglycerol and glycerophospholipids.
 HCO3- ,Acetyl-CoA, NADPH,
The cholesterol biosynthesis
The carbon origins of cholesterol as revealed by
radioisotope labeling studies.
 20. The cholesterol biosynthesis
pathway can be divided into four stages
Stage I: three acetyl-CoA molecules condense to form the 6-
carbon mevalonate ( 甲羟戊酸 ).
Stage II: mevalonate is converted to activated 5-carbon
isoprene ( 异戊二烯 ) units.
Stage III: Six isoprene units condense to form the linear 30-
carbon squalene( 鲨烯 ).
Stage IV: The linear squalene is cyclized to form a four-ring
structure, which is eventually converted to the 27-carbon
cholesterol through a series of complicated reactions.
(2C) Reactions
assembling
(6C) cholesterol from
18
(5C) molecules of
acetyl-CoA
(30C)
can be divided
into four
stages.

(27C)
 25. Cholesterols made in vertebrate
livers can be converted to bile acids and
cholesterol esters before exporting
 Cholesterol can be converted to bile acids and bile
salts, which will be secreted to the intestine for
emulsifying lipids.
 Cholesterol can also be converted to the more
hydrophobic cholesterol esters, which will be
stored in the liver or transported to other tissues
after being incorporated into lipoprotein particles.
 Cholesterol can be converted
to bile acids (salts) ：
glycocholate and taurocholate

牛黄胆酸盐

（甘胆酸盐）
Acyl-CoA-Cholesteryl
acyl transferase
(ACAT) catalyzes the
addition of an acyl
group to the hydroxyl
group of cholesterol
 29. All steroid hormones are derived
from cholesterol
Progesterone ( 孕酮 ) is synthesized from pregnenolone
by oxidizing the 3-OH group and the isomerization of
the double bond (from ∆ 5 to ∆ 4 position).
Cortisol (a major glucocorticoid) is synthesized from
progesterone by hydroxylation at C-17, C-21, and C-11.
Aldosterone (a mineralocorticoid) is synthesized from
progesterone by hydroxylation at C-21, C-11, and
oxidation of C-18 to an aldehyde.
 Testosterone (an androgen, or male hormone) is
synthesized from progesterone by the removal of 2-
carbon unit and hydroxylation at C-17.
 Estradiol ( an estrogen, or female hormone) is
synthesized from testosterone by the removal of C-
19 and formation of the aromatic A ring.

hypercholesterolemia and atherosclerosis

 27. The de novo synthesis of
cholesterol is regulated to complement
dietary intake
 HMG-CoA reductase, catalyzing the rate-limiting step of the
de novo cholesterol synthesis, has an activity variable over
100 fold!
 Hormones (insulin and glucagon) regulate the activity of the
HMG-CoA reductase via reversible phosphorylation.
 Genetic defect of the LDL receptor was found to cause the
familial hypercholesterolemia and atherosclerosis: the LDL
cholesterols can not enter the cells, while de novo synthesis
continues despite the high cholesterol level in the blood.
The de novo synthesis of
cholesterol is regulated
to complement the
dietary uptake

(For storage)
  Summary
all the 27 carbons of cholesterol are from
acetyl-CoA.
The biosynthesis of cholesterol takes a long
pathway, with the reaction catalyzed by HMG-
CoA reductase being the rate-limiting step for
de novo synthesis of cholesterol.
Activated isoprene untis, mevalonate,
squalene were found to be important
intermediates of cholesterol biosynthesis.
 26. Lipids (including cholesterols) are
transported in the vertebrate plasmia as
various lipoprotein particles
The different lipoprotein particles, having different
combinations of lipids and apolipoproteins, can be
separated by untracentrifugation due to different densities
and sizes.
The human plasma lipoproteins include chylomicrons
(which transports lipids from intestine to various tissues),
VLDL (very low density lipoproteins), LDL(Low density
lipoproteins), HDL (high density lipoproteins).
Lipids are transported
as various lipoprotein
particles in vertebrate
plasma
At least nine apolipoproteins (named as apo A, B, C, D, E) have
been revealed in human, which act as signals to target the
lipoprotein particles to various tissues or activating enzymes that
will act on the lipoproteins.
Endogenous lipids made in liver are transported to other tissues as
VLDL particles (~ 87% lipids and 12% proteins).
The apoC-II protein in VLDL activates the lipoprotein lipase in
muscle and adipocyte tissues, thus releasing free fatty acids there.
Some VLDL remnants is then converted to LDL (with 23% proteins
and 75% lipids) and with the others being absorbed by the liver cells
via receptor-mediated endocytosis.
LDL delivers cholesterols to extrahepatic tissues, where its apoB-100
protein (4636 residues) is recognized by specific LDL receptors and
LDL is endocytosed.
HDL, with its precursors formed in the liver or intestine cells, collects
the cholesterols in the plasma and deliver them to the liver cells.
(A negative correlation between blood HDL level and arterial diseases
has been observed.)
 L is uptaken by cells
a the LDL receptors,

LDL receptors
are recycled
to the cell surfaces
  Summary
The lipids are transported as lipoprotein
particles (including chylomicrons, VLDL,
LDL, and HDL).
The de novo biosynthesis of cholesterol is
regulated to complement the dietary
uptake.
All streroid hormones are derived from
choleterol.
A huge arrays of isoprenoids are made
using activated isoprene units.