Regulation of Fatty Acid Oxidation and Ketone Body Production

 This takes place in the liver where the fatty acyl-CoA formed in the cytoplasm may either
be oxidized in the mitochondria or converted into triglycerols of phospholipids by
enzymes in the cytoplasm. Fatty acyl-CoA are transferred from the cytoplasm to the
mitochondrial matrix via carnitine and once fatty acyl groups enter the mitochondria, they
are oxidized to acetyl-CoA.
 So carnitine controls the rate of fatty acid oxidation. Carnitine acyl transferase is an
allosteric enzyme which inhibited by its modulator, malonyl-CoA, an intermediate in the
biosynthesis of long when there is large amount of carbohydrate intake, because the
excess glucose which cannot be oxidized or stored as glycogen is converted into
triglycerols for storage. This occurs due to the allosteric inhibition of the entry of fatty
acyl groups into the mitochondria.
 In the mitochondria fatty acids may either be oxidized into CO2 and H2O via the citric
acid cycle or converted into ketone bodies which are carried to the peripheral tissues.
This is influenced by the availability of oxaloacetic acid which initiates the entry of acetyl
CoA into the citric acid cycle.
 When oxaloacetic acid concentration is low, only a small amount of acetyl-CoA enters
citric acid cycle, favoring ketone body production. There is low concentration of
oxaloacetic acid during fasting or low carbohydrate intake. Fatty acid oxidation is
increased with formation of acetyl-CoA, which is converted via hydroxymethyl glutaryl
CoA into aceto acetic acid, hydroxybutyric acid, butyric acid acetone. These ketone
bodies are carried to peripheral tissues where they are oxidized into CO2 and H2O via
the citric acid cycle.
Energy Conservation:

 The activation of fatty acids requires two ATP, so there is a deficit of 2 ATP per mole of
fatty acid oxidized. Each cleavage of a carbon-carbon bond yields 5 ATP. The acetyl
CoA produced is metabolized by the citric acid cycle to CO2 and H2O with production of
12 ATP for each acetyl CoA oxidized. A 16 carbon palmitic acid therefore will have the
following ATP yield: 7 cleavages ( 7 × 5 = 35 ) ; 8 acetyl CoA oxidized by citric acid
cycle ( 8 × 12 = 96 ) ; 2 ATP needed for activation; the total will be 35 + 96 = 131 – 2 =
129. Since 8000 cal. or 8 kcal. is needed for each ATP, then 129 × 8 kcal. = 1032 kcal.
Complete oxidation of palmitic acids yields theoretically 2340 kcal; 1032 ÷ 2340 × 100 =
44 % of standard free energy of oxidation of palmitic acid is recovered as ATP.
Lipogenesis:

 Due to the limited capacity of higher animals to store polysaccharides, any amount of
glucose ingested in excess of caloric requirement and storage capacity is converted to
fatty acids. Subsequently the fatty acids are synthesized into high caloric triglycerides
which are stored in the adipose tissues. From the quantitative point of view this is a
minor metabolic pathway, but because of the large amount of steroids derived from
cholesterol, the pathway of great significance.
 Lipogenesis occurs mostly in the liver, although there are evidences to support that other
tissues and even adipose tissue itself, contribute to the process.
 Any substance capable of yielding acetyl CoA is a potential source of carbon atoms for
the synthesis of fatty acids (Lipogenesis). There are three different systems for the
synthesis of fatty acids, all of which involve acetyl CoA, namely:

1. Cytoplasmic or de novo Synthesis of Palmitic Acid: (Non-mitochondrial)

 The complete de novo synthesis of long chain saturated fatty acids takes place only in
the soluble fraction of the cytoplasm. It is catalyzed by a complex of seven enzymes, the
fatty acid synthetase complex. Once molecule of acetyl CoA and seven molecules of
palmitic acid. The reducing power is supplied by NADPH.
CH3COCoA + 7COOHCH2COCoA + 14 NADPH + 14H +

acetyl CoA malony CoA

C15H31COOH + 7CO2 + 8CoA + 14NADP + 6H2O

Palmitic acid
 Acetyl CoA serves as primer. Its methyl and carboxyl carbon atoms become the 16 and
15 carbon atoms respectively of the palmitic acid. The lengthening of the chain starts at
the carboxyl group of acetyl CoA and proceeds by successive addition of acetyl residues
at the carboxyl end of the growing chain.
 Each successive acetyl residue is derived from the 2 carbon atoms of malonyl CoA
nearest the CoA group, the unesterified carboxyl group (3rd carbon) is lost as CO2. The
free carboxyl group of each residue of the entering malonyl CoA is displaced by the
carboxyl carbon atom of acyl CoA undergoing lengthening with formation of CO2 and b-
hydroxy fatty acyl derivative. This subsequently undergoes dehydration to unsaturated
fatty acyl derivative which is reduced to saturated fatty acyl residue by a second
molecule of NADPH. The cycle is repeated six or more times with the final formation of a
molecule of palmitic acid.
 The mechanism of fatty acid synthesis is distinct in that the acyl intermediates are not
thioesters of CoA as in fatty acid oxidation but are thioesters of small molecular weight
protein called acyl carrier protein (ACP). This protein can form complexes with one or
more of the six other enzyme proteins needed for the complete synthesis of palmitic
acid.
 If acetoacetyl CoA were to be formed from acetyl CoA alone by the action of thiolase,
the reaction would be strongly endergonic and its equilibrium would lie far to the left. On
the other hand the formation of aceto acetyl-S-ACP is strongly exergonic. Its equilibrium
lies far for the right, in the direction of synthesis. Thus the decarboxylation of the malonyl
residue provides a strong thermodynamic pull toward fatty acid synthesis.
 Formation of Malonyl CoA: The ultimate source of all the carbon atoms of palmitic acid
synthesized by cytoplasmic fatty acid synthetase complex is cytoplasmic acetyl CoA.
This is derived from the intramitochondrial acetyl CoA by indirect route. Since acetyl CoA
cannot pass through the mitochondrial membrane, it is first transformed to carnitine
which can pass the mitochondrial membrane to the cytoplasm. Subsequently the acetyl
group is transferred to cytoplasmic CoA.
COOH

CH2 + ADP + Pi
Mn + +
CH3COCoA + CO2 + ATP O = C SCoA
Acetyl CoA acetyl CoA malonyl
Carboxylase coA

 The malonyl CoA, immediate precursor of 14 of the 16 carbon atoms of palmitic acid is
formed by cytoplasmic acetyl CoA and CO2 through the catalyzing action of acetyl CoA
carboxylase. The carbon atom of CO2 becomes the distal or free carboxyl carbon of
malonyl CoA.
 Acetyl CoA carboxylase contains biotin as its prosthetic group bound in amide linkage to
a lysine residue of enzyme protein, biocytin. Biotin is essential for the reaction as shown
by inhibition by avidin.

2. A mitochondrial and 3. Microsomal Elongation of Saturated Fatty Acids:

 Neither the mitochondria nor the endoplasmic reticulum, represented by the microsome
fraction is capable of de novo synthesis of palmitic acid. However, they contain enzymes
capable of elongating the chains of fatty acids which already posses 12 to 16 carbon
atoms.
 Palmitic acid, the end product of cytoplasmic fatty acids synthetase system, may be
lengthened by two different types of enzymes systems; one in the mitochondria and the
other in the endoplasmic reticulum (microsomes).
 In the mitochondria, there is an enzyme which catalyzes the elongation of the preformed
saturated fatty acid (palmitic acid) by successive addition of acetyl CoA units. The
mitochondrial systems does not require CO2 fixation and the products are C18, C20, C22, C24
fatty acids. This pathway is essential the reverse of the oxidation of fatty acids, except
the reduction of a-b-double bond to form the saturated fatty acid which occurs at the
expense of NADPH as reductant. Mitochondrial system will also elongate unsaturated
fatty acids. There are four steps.
 Thiolase
a.CH3COCoA + RCH2COCoA RCH2COCH2COCoA + CoA
b-hydroxy
b. RCH2COCH2COSCoA + NADH + H + RCH2CHOHCH2COSCoA + NA
acyl dehydrogenase
enoyl
c. RCH2CHOHCH2COSCoA RCH2CH = CHCOSCoA + H2O
hydrase b-unsaturated acyl derivative
enoyl
d. RCH2CH = CHCOSCoA + NADPH + H + RCH2CH2CHCOSCoA
reductase

 This system is active not only on intermediate chain length saturated fatty acids C12, C14,
C16 but also on unsaturated compounds, lengthening the 2 C atoms at each step.
 The synthesis of fatty acid is distributed by : 1. Insufficient dietary intake. 2. Thiamine
deficiency, since thiamine pyrophosphate plays an important role in transforming
pyruvate into acetyl CoA.
 Microsomal lengthening of both saturated and unsaturated fatty acyl CoA esters utilizes
the malonyl pathway rather than the acetyl CoA. The fatty acid is first converted to acyl
CoA. Acyl CoA reacts with malonyl CoA followed by the reduction of the a-b-double
bond utilizing NADPH. The intermediates are similar to those in the de novo synthesis
but does not employ ACP as acyl carrier.
Fatty Acid Transformations:

 Fatty acids are taken from the diet and by lipogenesis from acetyl CoA. The composition
of fatty acid mixture in the diet vary in degree of saturated and chain length. Lipogenesis
favors the formation saturated over unsaturated as indicated by the hardening effect of
high carbohydrate diet, upon the fat depot. The liver produces fatty acid mixture
characteristics of the species, from out of the fatty acid mixture available.

a. Shortening and Lengthening of C skeleton:

 Isotopic studies has shown that shortening and lengthening of C skeleton of saturated
fatty acid occur in the animal body and that it takes place by gain or loss of 2 atoms at a
time.
b. Formation of Monoenoic Acid:
 The body is capable of dehydrogenating saturated fatty acids, to give 9,10 unsaturated
derivatives. When isotopic palmitic or stearic acid is fed, isotope can be recovered both
as saturated and unsaturated fatty acids, palmitoleic and oleic acid respectively.
 On the other hand, feeding with labeled oleic or palmitoleic acids showed that only small
amounts of saturated fatty acids are formed.
c. Formation and Transformation of Polyenoic Acids:
 A great variety of polyunsaturated fatty acids in the tissues includes:

 (1) Those formed by the animal, de novo. Those in which all double bonds lie between
the seven C from the terminal methyl and COOH group. These are produced by
alternate desaturation and chain elongation, Example: oleic acid and palmitoleic acid.
 (2) Those which cannot be made de novo. Here one or more of the double bonds are
situated within the terminal seven C atoms. These polyunsaturated fatty acids are
therefore indispensable in the diet, hence the term essential fatty acids. Example :
linolenic and arachidonic acid.
 Children with serum lipid low in essential fatty acids, developed eczematous lesions of
the skin. Administration of linoleic, linolenic and arachidonic acids clears up the lesion. In
this connection, pyridoxine deficiency has also been noted to produce similar lesions.
 And there seems to be a close relations between the vitamin and the unsaturated fatty
acids, because administration of either, cures the condition.
 Essential fatty acids exert favorable effects on sex maturation, pregnancy and lactation.
They increase capillary resistance and lower capillary permeability. They also play a role
in the metabolism.
 There are four groups of polyunsaturated fatty acids in man. Two are derived from
dietary linoleic and linolenic acids and two from mono unsaturated oleic and palmitoleic
acids, which are in turn derived from the corresponding saturated fatty acids. These four
groups can be recognized by the distance between the terminal methyl group and the
nearest double bond.