Fatty acid , PL , TG SYNTHESIS

Prof. Dalia Dasgupta

Mandal
Department of
Biotechnology
NIT Durgapur
 Lipid metabolism

Catabolis
m

Anabolis
m

Degradation Synthesis
Most tissues;
Tissue especially high in Mostly liver and a little
location liver, muscle and bit adipose
adipose
Subcellular Mitochondrial
Cytosol
location matrix

Electron NAD+ is electron

NADPH is electron donor
situation acceptor
1 ATP utilized for every 2
2 ATP used for carbons incorporated
activation of acyl (but obviously the
Energetics group, 108 reaped molecules being
for net 106 ATP incorporated are already
high-energy)
 Some facts about Fatty acid anabolism

• Acetyl-CoA is diverted from the citric acid cycle towards

fatty acid synthesis when high-energy molecules (ATP,
NADH) are abundant.

• Acetyl-CoA produced in mitochonrdria has to be

transported to the cytosol via the tricarboxylate
transport system for fatty acid synthesis.

• Reduction process that builds up the hydrocarbon chain

of fatty acids utilizing activated acyl unit and malonyl
unit Occurs in the cytoplasm

• Catalyzed by fatty acid synthase containing 7 catalytic

sites

• Incorporates carbon atoms from acetyl CoA into the

growing fatty acid chain

• Driven by the release of CO2 in a decarboxylation step

• Energetically unfavourable process

 Citrate shuttle : acetyl CoA transporter

Acetyl CoA is
synthesized in
the matrix of the
mitochondria
but fatty acids
are synthesized
in the cytosol.
How acetyl CoA
is made
available in the
cytoplasm?????
 Acetyl CoA (water soluble)  citrate (1st reaction of the TCA cycle)
 In the cytosol, citrate undergo the reverse reaction producing
oxaloacetate and acetyl CoA
 If the conditions favor, FA synthesis begins
 [ATP] allosterically inhibits isocitrate dehydrogenase (isocitrate is
not converted to α-ketoglutarate & TCA stops) Builds up isocitrate,
convert to citrate in the matrix
Regeneration of Acetyl Co A in the cytoplasm
Fig: Structure of fatty acid synthase
FATTY ACID SYNTHESIS
Overview of anabolic processes
 BIOSYNTHESIS OF TRIGLYCEROLS
 Triacylglycerols are formed by reaction of two molecules of fatty acyl–CoA with glycerol 3-
phosphate to form phosphatidic acid;
 This product is dephosphorylated to a diacylglycerol, then acylated by a third molecule of
fatty acyl–CoA to yield a triacylglycerol.
 The synthesis and degradation of triacylglycerol are hormonally regulated.
 Mobilization and recycling of triacylglycerol molecules results in a triacylglycerol cycle.
 Triacylglycerols are resynthesized from free fatty acids and glycerol 3-phosphate even during
starvation.
 The dihydroxyacetone phosphate precursor of glycerol 3-phosphate is derived from pyruvate
via glyceroneogenesis.
Biosynthesis of phosphatidic acid:
A fatty acyl group is activated by
formation of the fatty acyl–CoA, then
transferred to ester
linkage with L-glycerol 3-phosphate,
formed in either of the two ways
shown. Phosphatidic acid is shown
here with the correct stereochemistry
at C-2 of the glycerol molecule.
 Regulation of triacylglycerol synthesis by
insulin: Insulin stimulates conversion of dietary
carbohydrates and proteins to fat. Individuals
with diabetes mellitus lack insulin; in
uncontrolled disease, this results in diminished
fatty acid synthesis, and the acetyl-CoA arising
from catabolism of carbohydrates and proteins
is shunted instead to ketone body production.
Phosphatidic acid in lipid People in severe ketosis smell of acetone, so the
biosynthesis. Phosphatidic acid is condition is sometimes mistaken for
the precursor of both triacylglycerols drunkenness
The triacylglycerol cycle. In mammals, triacylglycerol molecules are broken down
and resynthesized in a triacylglycerol cycle during starvation. Some of the fatty
acids released by lipolysis of triacylglycerol in adipose tissue pass into the
bloodstream, and the remainder are used for resynthesis of triacylglycerol. Some of
the fatty acids released into the blood are used for energy (in muscle, for example),
and some are taken up by the liver and used in triacylglycerol synthesis. The
triacylglycerol formed in the liver is transported in the blood back to adipose tissue,
where the fatty acid is released by
extracellular lipoprotein lipase, taken up by adipocytes, and reesterified into
Glyceroneogenesis. The pathway is
essentially an abbreviated version of
gluconeogenesis, from pyruvate to
dihydroxyacetone phosphate (DHAP),
followed by conversion of DHAP to glycerol
3-phosphate, which is used for the
Regulation of glyceroneogenesis. (a)
Glucocorticoid hormones stimulate
glyceroneogenesis and gluconeogenesis
in the liver, while suppressing
glyceroneogenesis in the adipose tissue (by
reciprocal regulation of the gene expressing
PEP carboxykinase (PEPCK) in the two
tissues); this increases the flux through the
triacylglycerol cycle. The glycerol freed by
the breakdown of triacylglycerol in adipose
tissue is released to the blood and
transported to the liver, where it is primarily
converted to glucose, although some is
converted to glycerol 3-phosphate by
glycerol kinase.
(b) A class of drugs called thiazolidinediones
are now used to treat type 2 diabetes. In this
disease, high levels of free fatty acids in the
blood interfere with glucose utilization in
muscle and promote insulin resistance.
Thiazolidinediones activate a nuclear
receptor called peroxisome proliferator-
activated receptor (PPAR), which Induces the
activity of PEP carboxykinase.
Therapeutically, thiazolidinediones
increase the rate of glyceroneogenesis, thus
increasing the resynthesis of triacylglycerol
in adipose tissue and reducing the amount of
free fatty acid in the blood.
SYNTHESIS OF PHOSPHOLIPIDS
 Diacylglycerols are the principal precursors of
glycerophospholipids.
 In bacteria, phosphatidylserine is formed by the condensation of
serine with CDP-diacylglycerol;
 decarboxylation of phosphatidylserine produces
phosphatidylethanolamine.
 Phosphatidylglycerol is formed by condensation of CDP-
diacylglycerol with glycerol 3-phosphate, followed by removal of
the phosphate in monoester linkage.
 Yeast pathways for the synthesis of phosphatidylserine,
hosphatidylethanolamine, and phosphatidylglycerol are similar to
those in bacteria;
 phosphatidylcholine is formed by methylation of
phosphatidylethanolamine.
 Mammalian cells have some pathways similar to those in bacteria,
but somewhat different routes for synthesizing
phosphatidylcholine and phosphatidylethanolamine.
 The head-group alcohol (choline or ethanolamine) is activated as
the CDP derivative, then condensed with diacylglycerol.
Cells have two strategies to attach head
groups to phospholipids
Summary of the pathways to
phosphatidylcholine and
phosphatidylethanolamine.
Conversion of
phosphatidylethanolamine
to phosphatidylcholine in mammals
takes place only in the liver.
Overview of Cholesterol biosynthesis
Four stages:
Stage 2 : Mevalonate to Isoprene units
Stage 3 : SQUALENE
From Squalene to
Cholesterol
 Catabolism of FA
 Beta acyl
 The formation of fatty oxidation
CoA molecule prepares
fatty acids for entry into the mitochondria.
 Carnitine helps fatty acly CoA to enter
mitochondria. There they are degraded in the
catabolic process called beta oxidation.
 During beta oxidation, the third (or beta) carbon of
the saturated fatty acid chain of the fatty acyl CoA
is oxidized to a ketone.
 Beta oxidation is a spiral pathway. Each round
consists of four enzyme-catalyzed steps that yield
one molecule of acetyl CoA and an acyl CoA
shortened by two carbons, which becomes the
starting substrate for the next round.
 Seven rounds of beta oxidation degrade a C16 fatty
acid to eight molecules of acetyl CoA.
 Complete oxidation of one molecule of palmitic
acid to carbon dioxide and water yields 129
molecules of ATP. One round of beta oxidation
yields 17 ATP.
 Beta Oxidation is regulated by availability of free
The reactions of ß oxidation
• FAD accepts hydrogens from a fatty acyl-CoA in the first
step. A double bond is produced between the α- and β-
carbons, and an enoyl-CoA is formed. The FADH2 that is
produced interacts with the electron transport chain,
generating ATP.
• Enzyme: Acyl-CoA dehydrogenase (Multiple variants of this
enzyme)
• H2O adds across the double bond, and a β-hydroxyacyl-CoA
is formed.
• Enzyme: Enoyl-CoA hydratase
• β -Hydroxyacyl-CoA is oxidized by NAD+ to a β-ketoacyl-CoA.
The NADH that is produced interacts with the electron
transport chain, generating ATP.
• Enzyme: L-3-hydroxyacyl-CoA dehydrogenase (which is
specific for the L-isomer of the β-hydroxyacyl-CoA).
• The bond between the alpha and beta carbons of the β-
ketoacyl-CoA is cleaved by a thiolase that requires coenzyme
A. Acetyl-CoA is produced from the two carbons at the
carboxyl end of the original fatty acyl-CoA, and the
remaining carbons form a fatty acyl-CoA that is two carbons
shorter than the original.
• Enzyme: β -ketothiolase
Ketogenesis
• Ketogenesis results from β-oxidation of fatty acids in the hepatic
mitochondria, which is regulated by the activity of carnitine
palmitytol transferase I, which catalyzes uptake of fatty acid acyl-
CoA into the mitochondria.
• Ketone bodies are not obligately produced from fatty acids, but
rather any meaningful amount of them is synthesized only in a
situation of carbohydrate and protein insufficiency, where fatty acids
are the only readily available fuel for their production.
• The three ketone bodies, each synthesized from acetyl-CoA
molecules, are:
1. Acetoacetate, which can be converted by the liver into β-
hydroxybutyrate, or spontaneously turn into acetone. Most
acetoacetate is reduced to beta-hydroxybutyrate, which serves to
additionally ferry reducing electrons to the tissues, especially the
brain, where they are stripped back off and used for metabolism.
2. Acetone, which is generated through the decarboxylation of
acetoacetate, either spontaneously or through the enzyme
acetoacetate decarboxylase. It can then be further metabolized
either by CYP2E1 into hydroxyacetone (acetol) and then via
propylene glycol to pyruvate, lactate and acetate (usable for
energy) and propionaldehyde, or via methylglyoxal to pyruvate and
lactate.
3. β-hydroxybutyrate (not technically a ketone according to IUPAC
nomenclature) is generated through the action of the enzyme D-β-
hydroxybutyrate dehydrogenase on acetoacetate. Upon entering the
tissues, beta-hydroxybutyrate is converted by D-β-hydroxybutyrate
dehydrogenase back to acetoacetate along with a proton and a