BIOSYNTHESIS OF SATURATED FATTY ACIDS

(ASSIGNMENT # 2 SPRING 2022)

Submission Date (DEC 19, 2022)

BY

NIMRA AHKTAR

ROLL # 20011514-111

ZOO-476 (Biochemistry-II)

BS Zoology Semester 5th Section A

Submitted To

MISS RAZIA

Department of Zoology

UNIVERSITY OF GUJRAT

Session 2020-2024

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 TABLE OF CONTENTS
CONTENTS PAGE
LIST OF FIGURES ………………………………..…………………… 03
1. PROCESS OF BIOSYNTHESIS …………………………………….... 04
2. ACETYL COENZYME A ……………………………………………… 04
3. SYNTHESIS OF MALONYL COENZYME A………………………… 05
4. FATTY ACID SYNTHASE COMPLEX (FASC) DIMER ……………. 06
5. STEPS IN BIOSYNTHESIS OF SATURATED FATTY ACIDS ……… 06
5.1 : Condensation ………………………………………………………. 06
5.2: Reduction …………………………………………………………. 07
5.3: Dehydration ………………………………………………………. 07
5.4: Reduction …………………………………………………………. 07
5.5: Repeat ……………………………………………………………… 07
6. FATTY ACID ELONGATION………………………………………… 08
7. CONVERSION OF CARBOHYDRATES INTO FATTY ACIDS……. 8/9
8. REFERENCES………………………………………………………… 10

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 TABLE OF FIGURES
ii CONTENTS PAGE
Figure 01: Activation steps involved in acetyl CoA ……………………. 05
Figure 02: Acetyl CoA to Malonyl ……………………………………… 06
Figure 05: Pathway of steps involved in biosynthesis …………………. 07

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 BIOSYNTHESIS OF SATURATED FATTY ACIDS
1: Process
 The de novo synthesis of saturated fatty acids, studied in a variety of biological systems, is
catalyzed by two enzyme systems that function sequentially, acetyl-CoA carboxylase and fatty
acid synthetase. These enzyme complexes are found in the cell cytoplasm when cells are ruptured
and the cellular components are fractionated by usual techniques.
 The mechanism of action of acetyl-CoA carboxylase demonstrated that the enzymes contain
covalently bound biotin as a prosthetic group and that a carboxybiotin intermediate is formed
during the reaction. The mode of binding of the carboxyl group to the biotin was demonstrated by
studying a model reaction, the carboxylation of free biotin by β-methylcrotonyl-CoA
carboxylase; the product of this reaction was methylated and identified as 1'-N-
carboxymethylbiotin methyl ester.
 Acetyl-CoA carboxylase is the first committed reaction in the biosynthesis of fatty acids, and this
enzyme is known to be subject to two types of control, allosteric regulation and adaptive changes
in enzyme content. (Albert, 1999)

2 : Acetyl Coenzyme A:
 Coenzyme A consists of and chain made up of ADP, pantothenic acid, and triethanolamine, with
an H2S group on the end which binds the acetate group of acetyl CoA.
 Acetyl CoA is produced in the matrix of the mitochondria, but fatty acid biosynthesis occurs in
the cytosol.
 Citrate synthase frees CoA from acetyl CoA and condenses acetate and oxaloacetate to citrate.
 Matrix membrane transporters for citrate move citrate to the cytosol, where it is acted upon by
citrate lyase in the presence of CoA to re-form acetyl CoA and oxaloacetate.
 The oxaloacetate produced is converted to malate, and then to pyruvate, which is transported back
to the mitochondrial matrix.
 The conversion of malate to pyruvate releases NADPH into the cytosol, which is necessary for
fatty acid biosynthesis. (The hexose monophosphate shunt, pentose phosphate pathway, is the
other major source for cytosolic NADPH). (Smith,2001)

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 Figure 02 : Activation steps invloved n acetyl COA

(Vhope,2008)

3 : Synthesis of Malonyl Coenzyme A

 Acetyl CoA, with the addition of CO2, and with the hydrolysis of an ATP, is converted to
malonyl CoA by acetyl CoA carboxylase (a biotin-dependent enzyme like all carboxylases).
 Acetyl CoA carboxylase (ACC) is, being the first enzyme in the fatty acid biosynthetic pathway,
is a regulated enzyme.
 In the short term, allosteric activation by citrate, and allosteric inactivation by malonyl and
palmitoyl CoAs, and covalent modification (phosphorylation and dephosphorylation) are the
principal regulatory mechanisms.
 ACC is normally present as a tetrameric protomer (inactive form). The active form is the large
polymer, which is favored by citrate binding and inhibited by malonyl and palmitoyl CoAs
(products of the FA biosynthetic pathway).
 Phosphorylation is regulated by another mechanism, with glucagon and epinephrine activating
PKA to phosphorylate (inactivate) ACC, and insulin activating phosphatase to re-activate the
enzyme.
 The burden of long-term regulation is carried almost exclusively by up regulating the
transcription of the enzyme itself (Daienl,2006)

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Figure 03 : Acetyl COA to Malonyl COA

(Micheal , 2004)

4 : Fatty Acid Synthase Complex (FASC) Dimer

 Seven enzymes and a “carrier” protein: acetyl CoA-ACP transacylase, malonyl CoA-ACP
transacylase, β-ketoacyl-ACP synthase (condensing enzyme), β-ketoacyl-ACP reductase, β-
hydroxyacyl-ACP dehydratase, enoyl-ACP reductase, palmitoyl thioesterase, and acyl carrier
protein (ACP) (containsephosphopentetheine)
 The sulfhydryl group of one ACP unit associates with the enoyl-ACP reductase (ER) subunit of
another FASC complex, allowing dimerization of the protein.
 ACP assists in reactions by binding to substrate molecules, such as acetate (from acetyl CoA) and
malonate (from malonyl CoA).
 Any time a fatty acid is used in a biosynthetic reaction in the cell, it must be in the form of a fatty
acyl CoA. (Fukunaga,1995)

5 : Steps in Biosynthesis of Saturated Fatty acids

5.1 : Condensation
 Acetate (2C) and malonate (3C), as acetyl-ACP and malonyl-ACP
 Releases the non-ACP-bound carboxyl group of malonate as CO2
 Produces β-acetoacetyl-ACP (4C) (Lubert,2016)

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5.2 : Reduction
 Produces β-hydroxybutyryl-ACP (4C), with the oxidation of NADPH2 to NADP+
5.3 : Dehydration
 Produces crotonyl-ACP (4C) with the release of water
5.4 : Reduction
 Produces Butyryl-ACP with the oxidation of NADPH2 to NADP+
5.5 : Repeat
 Butyryl-ACP then enters into reaction 1 in the place of malonyl-ACP, undergoing the addition of
another two carbons from acetate.
 The overall reaction uses 8 acetyl CoA, 14 NADPH, 14 H+ and 1 malonyl CoA to produce a 16-
carbon palmitic acid. (Morita,2014)

Figure 05 : Pathway of steps involved in bosynthesis

(Eric , 2002)

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6 : Fatty Acid Elongation:
 FASC produces palmitic acid (16C).
 In the endoplasmic reticulum, two-carbon units can be added to palmitate from malonyl CoA.
 In the mitochondria, two-carbon units can be added to 8C fatty acids from acetyl CoA, but only to
the extent of 14C fatty acids. (Gotoh,2005)

7 : Conversion of carbohydrates into fatty acids

 In humans, fatty acids are formed from carbohydrates predominantly in the liver and adipose
tissue, as well as in the mammary glands during lactation.
 The pyruvate produced by glycolysis is an important intermediary in the conversion of
carbohydrates into fatty acids and cholesterol. This occurs via the conversion of pyruvate into
acetyl-CoA in the mitochondrion. However, this acetyl CoA needs to be transported into cytosol
where the synthesis of fatty acids and cholesterol occurs. This cannot occur directly.
 To obtain cytosolic acetyl-CoA, citrate (produced by the condensation of acetyl CoA with
oxaloacetate) is removed from the citric acid cycle and carried across the inner mitochondrial
membrane into the cytosol. There it is cleaved by ATP citrate lyase into acetyl-CoA and
oxaloacetate. The oxaloacetate can be used for gluconeogenesis (in the liver), or it can be
returned into mitochondrion as malate. The cytosolic acetyl-CoA is carboxylated by acetyl CoA
carboxylase into malonyl CoA, the first committed step in the synthesis of fatty acids.(
Okuyama,1994 )

8: Regulation
 Acetyl-CoA is formed into malonyl-CoA by acetyl-CoA carboxylase, at which point malonyl-
CoA is destined to feed into the fatty acid synthesis pathway. Acetyl-CoA carboxylase is the
point of regulation in saturated straight-chain fatty acid synthesis, and is subject to both
phosphorylation and allosteric regulation. Regulation by phosphorylation occurs mostly in
mammals, while allosteric regulation occurs in most organisms.
 Allosteric control occurs as feedback inhibition by palmitoyl-CoA and activation by citrate.
When there are high levels of palmitoyl-CoA, the final product of saturated fatty acid synthesis, it
allosterically inactivates acetyl-CoA carboxylase to prevent a build-up of fatty acids in cells.
Citrate acts to activate acetyl-CoA carboxylase under high levels, because high levels indicate
that there is enough acetyl-CoA to feed into the Krebs cycle and conserve energy.
 High plasma levels of insulin in the blood plasma (e.g., after meals) cause the dephosphorylation
of acetyl-CoA carboxylase, thus promoting the formation of malonyl-CoA from acetyl-CoA, and

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consequently the conversion of carbohydrates into fatty acids, while epinephrine and glucagon
(released into the blood during starvation and exercise) cause the phosphorylation of this enzyme,
inhibiting lipogenesis in favor of fatty acid oxidation via beta-oxidation (Zhang,2015)

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 REFERNCES
Dijkstra, Albert J.; Hamilton, R. J.; Hamm, Wolf (2008). " Fatty Acid Biosynthesis”
Voet, Donald; Voet, Judith G.; Pratt, Charlotte W. (2006). “Fundamentals of Biochemistry”
Stryer, Lubert (1995). "Fatty acid metabolism”
Wang, Haihong; ECronan, John (2004). “Straight chain fatty acids”
Naik DN, Kaneda T (December 1974). "Biosynthesis of branched long-chain fatty acids”

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