EVEN N ODD F.

• Fatty acids provide highly efficient energy storage, delivering more energy per gram than
carbohydrates like glucose.
• In 1904, franz knop gave the mechanism of fatty acid oxidation.
• The fatty acid are according to knop, F.A are degraded into 2 carbon units which is known as the
knoop hypothesis.

•
• OXIDATION OF EVEN-CHAIN SATURATED FATTY ACIDS ( = KNOOP’S β OXIDATION
PATHWAY)-
• β-Oxidation (in which all reactions involve the β-carbon of a fatty acyl-CoA) is a spiral
consisting of four sequential steps, the first three of which are similar to those in the TCA
cycle between succinate and oxaloacetate.
• Stage 1- Beta oxidation pathway
• First Stage : β oxidation pathway In this stage, the fatty acids undergo oxidative removal of
successive two-carbon units in the form of acetyl-CoA, starting from the carboxyl end of the
fatty acyl chain.
• Formation of each molecule of acetyl-CoA requires removal of 4 hydrogen from the fatty
acyl moiety by the action of dehydrogenases.
• Second Stage : Citric acid cycle.
• In this stage of fatty acid oxidation, the acetyl residues of acetylCoA are oxidized to CO2 via
the citric acid cycle, which also takes place in the mitochondrial matrix.
• Third Stage : Mitochondrial respiratory chain
• The first two stages of fatty acid oxidation produce the electron carriers, NADH and FADH2,
which in the third stage donate electrons to the mitochondrial respiratory chain, through
which electrons are carried to oxygen.The, energy released by fatty acid oxidation is
conserved as ATP.
• STEPS OF BETA OXIDATION=
•
Step 1 – alpha, beta oxidation of acyl-coa
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)
Step2- hydration α, β-unsaturated acyl-CoAs
H2O adds across the double bond, and a β-hydroxyacyl-CoA is formed.
• Enzyme: Enoyl-CoA hydratase
Third Step : Oxidation of β-hydroxyacyl-CoA
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: beta-hydroxyacyl-CoA dehydrogenase (which is specific for the L-isomer of the β-
hydroxyacyl-CoA).
• Fourth Step : Thiolysis
• 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
The shortened fatty acyl-CoA repeats these four steps. Repetitions continue until all the
carbons of the original fatty acyl-CoA are converted to acetyl-CoA.
• Energy Yield for Even-chain Fatty Acids
• The 16-carbon palmitoyl-CoA undergoes seven repetitions.
• In the last repetition, a 4-carbon fatty acyl-CoA (butyryl-CoA) is cleaved to two acetyl-CoAs.
• When one palmitoyl-CoA is oxidized, seven FADH2, seven NADH, and eight acetyl-CoA are
formed.
• The seven FADH2 each generate approximately 1.5 ATP, for a total of about 10.5 ATP.
• The seven NADH each generate about 2.5 ATP, for a total of about 17.5 ATP.
• The eight acetyl-CoA can enter the TCA cycle, each producing about 10 ATP, for a total of
about 80 ATP.
• From the oxidation of palmitoyl-CoA to CO2 and H2O, a total of about 108 ATP are
produced.
• The net ATP produced from palmitate that enters the cell from the blood is about 106
because palmitate must undergo activation (a process that requires the equivalent of 2 ATP)
before it can be oxidized (108 ATP − 2 ATP = 106 ATP).
• Oxidation of odd-chain and unsaturated fatty acids
• Odd-chain fatty acids produce acetyl-CoA and propionyl-CoA.
The odd-carbon long-chain fatty acids are oxidized by the same pathway as the even one.
However, the substrate for the last pass through the β oxidation cycle is a fatty acyl-CoA,
When this is oxidized and finally cleaved, the products are acetyl-CoA and propionylCoA,
rather than 2 moles of acetyl-CoA produced in the normal β oxidation cycle.
The acetyl-CoA is, of course, oxidized via the citric acid cycle but the oxidation of
propionylCoA appears to be a substrate unsuitable for β oxidation. However, the substrate
is held by two dissimilar pathways: methylmalonate pathway and β-hydroxy-propionate
pathway.
1. methylmalonate pathway
• This pathway is found only in animals and occurs in the mitochondria of liver, cardiac
and skeletal muscles, kidney and other tissues. Propionate (or propionyl-CoA) is also
produced by the oxidation of isoleucine, valine, methionine and threonine
 •
• Propionic acid is converted to propionyl co-A by acetyl CoA Synthetase
• propionyl co-A is carboxylased to methylmalonyl-CoA by an enzyme propionyl-CoA
carboxylase
• methylmalonyl-CoA is epimerized to L-methylmalonyl-CoA, by the action of
methylmalonyl-CoA epimerase
• L-methylmalonyl-CoA undergoes an intramolecular rearrangement to form succinylCoA
by the enzyme methylmalonyl-CoA mutase
2. β-hydroxypropionate Pathway
• This pathway is ubiquitous in plants
• Since plants have no B12 functional enzymes, the methylmalonate pathway does not
operate in them

•
• Two inheritable types of methylmalonic acidemia (and aciduria) are associated in young
children with failure to grow and mental retardness.
What are ketone bodies? Describe briefly what is meant by ketogenesis.
Ketogenesis is a catabolic pathway of metabolism. In this process, fatty acids and certain
ketogenic amino acids are broken down to derive energy by alternative means. Ketone bodies
are produced in the ketogenesis process.
Ketone Bodies
Fatty acids undergo 𝛽-oxidation in the liver mitochondria to generate a high amount of energy
and form three compounds, that are known as “ketone bodies”. These ketone bodies are
water-soluble and do not require lipoproteins for transportation across the membrane. Ketone
bodies are lipid molecules having a carbonyl group attached to two -R groups.
The three ketone bodies formed are:

1. Acetoacetate
2. D-3-hydroxybutyrate
3. Acetone

Ketogenesis Pathway

• Our body normally derives energy from stored carbohydrate by the process of
glycogenolysis (glycogen → glucose) or from non-carbohydrate sources such as lactate by
the process of gluconeogenesis.
• Ketogenesis occurs continuously in a healthy individual, but under certain conditions, when
there is an increased concentration of fatty acids or carbohydrate reserves are decreased,
ketogenesis happens at a higher rate:
• Under low blood glucose level, e.g. during fasting or starvation
• On exhaustion of carbohydrate reserve, e.g. glycogen
• When there is insufficient insulin, e.g. Type-1 diabetes
• All the main body parts such as the brain, skeletal muscles, heart, etc. can utilise the energy
formed by ketogenesis.
• Insufficient gluconeogenesis results in hypoglycemia and excessive production of ketone
bodies resulting in a fatal condition called ketoacidosis.
• Ketogenesis Steps
• The ketogenesis process occurs primarily in the mitochondria of liver cells. Below are the
steps in the process of ketogenesis:
• Transfer of fatty acids in mitochondria by carnitine palmitoyltransferase CPT-1
• 𝛽-oxidation of fatty acid to form acetyl CoA
• Acetoacetyl-CoA formation: 2 acetyl CoA form acetoacetyl CoA. The reaction is catalyzed by
the enzyme thiolase
• 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthesis: the step is catalyzed by HMG-CoA
synthase
• Acetoacetate formation: HMG-CoA is broken down to acetoacetate and acetyl-CoA by the
action of HMG-CoA lyase
• Acetoacetate thus produced forms other ketone bodies, acetone by decarboxylation and D-
3-hydroxybutyrate by reduction

•
• Liver, which produces ketone bodies, primarily in the mitochondria, cannot utilise it due to
lack of an enzyme 𝛽-keto-acyl-CoA transferase.
• Acetoacetate and D-3-hydroxybutyrate are used by the body to get energy. These ketone
bodies are circulated out of the liver cell.
• In the extrahepatic tissues, the following reactions occur:
• D-3-hydroxybutyrate is converted back to acetoacetate by 𝛽-hydroxybutyrate
dehydrogenase
• Acetoacetate is converted back to acetyl-CoA by 𝛽-keto-acyl-CoA transferase
• Acetyl-CoA enters the citric acid cycle (TCA or Kreb’s cycle) and produces 22 ATP molecules
• Acetone is excreted out
• Ketogenesis process is regulated by Insulin. Hormones such as glucagon, thyroid hormones,
catecholamines, cortisol increase ketogenesis rate by stimulating the breakdown of free
fatty acids.
• Significance of Ketogenesis
• Ketogenesis is used to get energy by the brain, heart and skeletal muscles under fasting
condition
• The ketogenic diet (low-carb, fat-rich diet) is used these days to lose weight. The idea is to
utilise excess fat stored in the body to get energy, but excess ketone bodies production can
lead to various complications and ketoacidosis
• In ketoacidosis condition, the kidneys excrete extra ketone bodies with the water resulting
in fluid loss
• Diabetic patients are greatly affected by ketoacidosis because insulin hormone is the main
regulator of the process
Q. Write in detail about biosynthesis of cholesterol and its regulation via synthesis
Q. Write in detail about biosynthesis of cholesterol and its regulation via synthesis.
Ans.

● This extremely difficult pathway was explained by Konrad Bloch, Feodor Lynen, John
Kornforth and George Popjáck.
● Its also referred to as the Mevalonate pathway.

●
● Most of the cholesterol synthesis takes place in the vertebrate liver cells
● Biosynthesis of cholesterol generally takes place in the endoplasmic reticulum of hepatic
cells.
● Like long-chain fatty acids, cholesterol is made from acetyl-CoA. The synthesis of
cholesterol takes place in 4 stages.
● Reaction:
● Stage 1: Conversion of 3 acetate units to mevalonate:
○ , 3 acetate units condense to form mevalonate, an intermediate of cholesterol
synthesis.
○ This in turn forms acetoacetyl-CoA by using thiolase, which then condenses with
a third mole of acetyl-CoA to yield HMG- CoA, using HMG-CoA synthase as an
enzyme.
○ These first two reactions are reversible.
○ The third reaction, i.e., the reduction of HMG-CoA to mevalonate, is a committed
step, wherein 2 moles of NADPH each donate two electrons. This reaction is
catalyzed by HMG-CoA reductase. It is the major point of regulation for
cholesterol synthesis.
○ It may, however, be pointed out that HMG- CoA is converted into mevalonate in
the cytosol, but it is converted into acetyl-CoA and acetoacetate in the
mitochondria.
● Stage 2 : Conversion of mevalonate to 2 activated isoprenes:
○ The three phosphate groups are transferred from 3 ATP molecules to
mevalonate.
○ The phosphate group attached to the C-3 hydroxyl group of mevalonate in the
intermediate compound, 3-phospho-5-pyro phosphomevalonate is a good
leaving group.
○ In the next reaction, this phosphate and the adjacent carboxyl group both leave,
thus producing a double bond in ∆3-isopentenyl pyrophosphate. This is the first
of the two activated isoprenes essential to cholesterol formation.
○ Isomerization of ∆3-isopentenyl pyrophosphate yields the second activated
isoprene, dimethylallyl pyrophosphate.
● Stage 3 : Condensation of 6 activated isoprenes to squalene
○ Isopentenyl pyrophosphate and dimethylallyl pyrophosphate now undergo a
‘head-to-tail’ condensation wherein one pyrophosphate group is displaced and
geranyl pyrophosphate is formed.
○ The ‘head’ is the end to which pyrophosphate is joined.
○ Geranyl pyrophosphate then undergoes another ‘head-to-tail’ condensation with
farnesyl pyrophosphate.
○ 2 moles of farnesyl pyrophosphate then undergo another ‘head-to tail’
condensation, forming squalene.
○ Squalene has 30 carbon atoms, 24 in the main chain and 6 in the form of methyl
group branches.
● Stage 4 : Conversion of squalene to the steroid nucleus
 ○ All of the sterols are alcohols with an OH group at C-3 and have a steroid nucleus
containing 4 fused rings.
○ One oxygen atom from oxygen is added to the end of the squalene chain forming
an epoxide.
○ The reaction is catalyzed by squalene monooxygenase, another mixed-function
oxidase enzyme.
○ NADPH reduces the other oxygen atom of oxygen to water.
○ The double bonds of the product epoxide are positioned so that the linear
squalene epoxide converts into a cyclic structure.
○ In animals, this cyclization results in the formation of lanosterol. Lanosterol is
finally converted into cholesterol in a series of about 20 reactions, including the
migration of some methyl groups and the removal of others.
● Significance:
○ Cholesterol fulfills a number of biological functions and is necessary for
successful cellular homeostasis.
○ It acts as a precursor to bile acids.
○ It also assists in steroid and vitamin D synthesis.
○ Plays a central role in maintaining cellular membrane rigidity and fluidity.
○ The cholesterol content and the rate of cholesterol biosynthesis are elevated in
growth of normal tissues and tumors.
○ Cholesterol biosynthesis happens much before DNA synthesis, and inhibiting
cholesterol biosynthesis inhibits cell growth, suggesting a linkage between the
cholesterol and DNA synthetic pathways.
ELONGATION OF F.A IN MITO N MICROSOME
• Palmitate (a C16 fatty acid) is the major product of the fatty acid synthase system.
• This system is also called as the de novo system in that the palmitate is constructed from
acetyl-CoA (ACP) and malonyl-CoA (ACP).
• In plants and animals, the most important fatty acids are the C18 fatty acids, namely
stearic, oleic, linoleic and linolenic.
• These C18 fatty acids are synthesized from the de novo system. Palmitate acts as a
precursor of other long-chain fatty acids. It may be lengthened to form stearate (18 : 0) or
even larger saturated fatty acids by further addition of acetyl groups through the action of
fatty acid elongation systems present in the microsomes and mitochondria in the case of
animals and in the soluble cytosol in the case of plants.
• PLANTS-
•
• Palmitate is the precursor of stearate and longer-chain saturated fatty acids, as well as the
monounsaturated fatty acids, palmitoleate and oleate.
• Mammals cannot convert oleate into linoleate, hence required in the diet as essential fatty
acids (EFAs).

Elongation: Mitochondrial Fatty Acid Elongation,

In the mitochondria fatty acid elongation occurs by a reversal of beta-oxidation.

This shows the overall reaction of fatty acid elongation in mitochondria. The process is
essentially a reversal of beta-oxidation, except that one NADPH and one NADH are required
(beta-oxidation yields two NADH). Mitochondrial fatty acid elongation acts primarily on fatty
acyl CoA substrates shorter than 16 carbons.
•

•
• Although different enzyme systems are involved and coenzyme A, is the acyl carrier directly
involved in animal fatty acid synthesis, the mechanism of elongation is otherwise identical
with that employed in palmitate synthesis: donation of two carbons by malonyl-ACP,
followed by reduction (with NADH), dehydration and another reduction (with NADPH) to
the saturated C18 product, stearyl-CoA. Fasting largely abolishes chain elongation.
• Elongation of stearyl-CoA in brain increases rapidly during myelination in order to provide
C22 and C24 fatty acids that are present in sphingolipids.

BIOSYNTHESIS N DEGRADATION OF TAG

• Depending upon the requirements of organisms, most of the fatty acids synthesized or
ingested by an organism are either incorporated into triacylglycerols for the storage of
metabolic energy or incorporated into the phospholipid components of membranes.
• (triacylglycerol pathway begin with the formation of fatty acyl esters of glycerol.
• TAG synthesis occurs in adipose tissues
• When carbohydrate is ingested in excess of the capacity to store glycogen, it is converted
into triacylglycerols and stored in adipose tissues. Plants also do manufactur triacylglycerols
as an energy-rich fuel and store them in fruits and seeds.

•
• Fatty acyl groups are first activated by formation of fatty acyl–CoA molecules, then they are
transferred to ester linkage with L-glycerol-3-phosphate, formed in either of the 2 ways
shown.

•
• STEPS-
• Triacylglycerols and glycerophospholipids (phosphatidylethanolamine, for example) share
two precursors (fatty acyl-CoAs and glycerol-3-phosphate) and many enzymatic steps in
their biosynthesis in animal tissues.
• Glycerol-3-phosphate can be formed in two ways (:
(a) either from dihydroxyacetone phosphate generated during glycolysis by the action of
• cytosolic NAD-linked glycerol-3-phosphate dehydrogenase,
(b) or from glycerol by the action of glycerol kinase in liver and kidney.
• Phosphatidate, which occurs in traces in cells, is a key intermediate in lipid biosynthesis.
• It can be converted either to a triacylglycerol or to a glycerophospholipid. In the pathway to
triacylglycerols, phosphatidate is hydrolyzed by phosphatidate phosphatase to form 1, 2-
diacylglycerol .
Diacylglycerols are then converted to triacylglycerols by transesterification with a third
fatty acyl-CoA.
• Significance-
• In men, the amount of body fat remains almost constant over long periods. However, if
carbohydrate, protein or fat is consumed in excess of their energy needs, the surplus is
stored in the form of triacylglycerols.
• This stored fat can be utilized for energy during normal body activity or during fasting.
Triacylglycerol metabolism is influenced by hormones such as insulin and
glucagon and also by the growth hormone and adrenalocorticoids.

Q. Write short notes on

a. Biosynthesis of p. Ethanolamine
b. Biosynthesis of p. Choline
c. Biosyntheisi of p. Serine
Interrelationship among the Eukaryotic Pathways to Phosphatidylserine,
Phosphatidylethanolamine and Phosphatidylcholine

● In yeast, as also in bacteria, phosphatidylserine may be formed by condensation of CDP-

diacylglycerol and serine.
● And phosphatidylethanolamine can be synthesized from phosphatidylserine in the
reaction catalyzed by phosphatidylserine decarboxylase
● Note that phosphatidylserine and phosphatidylethanolamine are interconverted by a
reversible head group exchange reaction. adoHcy = S-adenosylhomocysteine.
● An alternative pathway to phosphatidylserine is a head group exchange reaction, in
which free serine displaces ethanolamine. Phosphatidylethanolamine may also be
converted into phosphatidylcholine (lecithin) by the addition of 3 methyl groups to its
amino group.
● Mammals, however, do not synthesize phosphatidylserine from CDP-diacylglycerol.
Instead, they derive it from phosphatidylethanolamine via the head group exchange
 reaction, as depicted in Fig. 25–24. In mammals, the synthesis of all nitrogen-containing
phospholipids occurs by Strategy 2: phosphorylation and activation of the head group
followed by condensation with diacylglycerol. As an instance, choline is resused (or
“salvaged”) by first being phosphorylated and then converted into CDP-choline by
condensation with CTP. A diacylglycerol displaces CMP from CDP-choline, forming
phosphatidylcholine.
● Ethanolamine, obtained in the diet, is converted into phosphatidylethanolamine by an
analogous salvage pathway. In the liver, phosphatidylcholine is also produced by
methylation of phosphatidylethanolamine, using s-adenosylmethionine. In all other
tissues, however, phosphatidylcholine is produced only by condensation of
diacylglycerol and CDP-choline.