Lipid Metabolism

GC-2-MS 12.1 Digestion and Transport of Fat

(1427-108)
12.1 Fatty Acids Degradation
Dr. Ahmed Almehdi 12.1 Ketone Bodies (ketoacidosis)
12.1 Fatty Acids Biosynthesis

Biochemistry

Biochemistry, the molecular basis of life – McKee , 7th Ed (2020) , Oxford University Press
 12.1 Digestion & Transport

 acetyl-CoA: the energy-rich molecule composed of

coenzyme-A and the two carbon acetyl group,
plays an important role in the metabolism of lipids
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 acetyl-CoA: is the product of fatty acid

degradation and the precursor (substrate) for
fatty acid biosynthesis
 fatty acids: are an important and efficient energy
source for many cell
 after triacylglycerol molecules are ingested, they
are mixed with bile salts in the small intestine,
digested by pancreatic lipases into
monoacylglycerol + 2 fatty acids
 Cont. …12.1 Digestion & Transport
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 both monoacylglycerol and fatty acids are

transported into the intestinal cells, reconverted
back to triacylglycerol, joined with proteins to
form chylomicrons
 chylomicron: transports triacylglycerol to be
stored in the adipocytes
 Cont. …12.1 Digestion & Transport

 most of the triacylglycerol content in circulating chylomicrons is

removed by skeletal muscle and adipose tissue cells (adipocytes)
 Triacylglycerols: are constantly synthesized and hydrolyzed to fatty
acids and glycerol (triacylglycerol cycle)
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 triacylglycerol cycle: regulates the level of fatty acids available to the

body for energy generation
 net result is sufficient fatty acids available for the body’s energy and
biosynthetic requirements
 depending on the metabolic needs, fatty acids may be:
 converted to triacylglycerol
 degraded to generate energy
 used for membrane synthesis
 Cont. …12.1 Digestion & Transport

 the triacylglycerol cycle is a mechanism that regulates the level of

fatty acids available to the body for energy generation
 glycerol-3-P binds to 3 fatty acids to form triacylglycerol (TG)
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 Cont. …12.1 Digestion & Transport

 glyceroneogenesis: is the
synthesis of glycerol-3-P from
substrates other than glucose
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or glycerol (malate → OAA →

PEP → DHAP → glycerol-3-P)
 glyceroneogenesis:
occurs in the adipocytes
 Cont. …12.1 Digestion & Transport

 when energy reserves are low, the body’s fat (triacylglycerol)

stores are mobilized (degraded into glycerol + 3 fatty acids) in a
process termed lipolysis
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 lipolysis: in adipose tissue occurs during fasting, during vigorous

exercise, and in response to stress
 then fatty acids are transported to target organelle by fatty acid
binding proteins
 the pathway for the breakdown of fatty acids into acetyl-CoA is
called β-oxidation cycle and occurs in the matrix of mitochondria
 fatty acids are degraded by the sequential removal of two carbon
fragments from the carboxyl end of fatty acids as acetyl-CoA
 Cont. …12.1 Fattyacyl Transfer

 Fatty acid binds to Coenzyme-A forming

fattyacyl-CoA
 acyl groups (from fattyacyl-CoA) are
transported from the cytosol into the
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mitochondrial matrix by carnitine

(where β-oxidation cycle occurs)
 carnitine-mediated acyl group transfer is
accomplished by: 1) acyl-CoA converted into
acylcarnitine (catalyzed by acyltransferase- )
2) carrier protein (translocase) transfers
acylcarnitine into the matrix
3) acyltransferase- converts acylcarnitine to acyl-CoA
4) carnitine is recycled back to the cytoplasm
carnitine
 Cont. …12.1 β-oxidation cycle

β-oxidation cycle:
 degradation of fatty acids starts with binding to coenzyme-A
(activation, forming fatty acyl-CoA, and requires 2 ATP hydrolysis)
 each β-oxidation cycle: of saturated fatty acids is a series of four
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reactions (oxidation → hydration → oxidation → cleavage), that

generates: 1 FADH2 , 1 NADH , and 1 acetyl-CoA
 acetyl-CoA: is further degraded in the citric acid cycle into 2CO2 and
more NADH and FADH2 are generated
 NADH and FADH2 are oxidized in the in electron transport chain (ETC)
in mitochondria to produce ATP
 the shortened fattyacyl-CoA is further oxidized with additional cycles
until the entire chain is degraded into acetyl-CoA
 Cont. …12.1 β-oxidation cycle
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 Cont. …12.1 β-oxidation cycle

 carbohydrate storage in the body is

limited, while fat reserves are not
 complete aerobic oxidation of a
Oxidation of Stearic acid (18:0)
fatty acid generates a large number
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of ATP molecules
 Stearic acid degradation produces:
8 FADH2 (8 x 1.5 = 12 ATP) 1.5 12
+ 8 NADH ( 8 X 2.5 = 20 ATP) 2.5 20
+ 9 acetyl-CoA (90 ATP)
–2 ATP needed for activation
(total = 122 ATP)
2.5 67.5
 Palmitic acid degradation produces: 1.5 13.5
7 FADH2 + 7 NADH + 8 acetyl-CoA (80 122
ATP) (–2ATP) , (total 106 ATP)
 Cont. …12.1 β-oxidation cycle

ketoacidosis:
 most of the acetyl-CoA produced during fatty acid oxidation is used by the
citric acid cycle or in isoprenoid (cholesterol) synthesis
 under normal conditions, fatty acid metabolism is so carefully regulated
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that only small amounts of excess acetyl-CoA are produced

 excess acetyl-CoA molecules are converted to ketone bodies (acetoacetate,
β-hydroxybutyrate, and acetone), referred to as ketogenesis
 high ketone bodies in blood is referred to as ketoacidosis
 Ketogenesis occur in the matrix of liver mitochondria
 acetone (ketosis) is produced when acetoacetate levels is high during
starvation and uncontrolled diabetes
 cardiac and skeletal muscle use ketone bodies as energy (brain does during
prolonged starvation)
  some organs (heart and skeletal muscle)
can use ketone bodies (acetoacetate
and β-hydroxybutyrate) as an energy
source under normal conditions
 during starvation, the brain uses ketone
bodies as source of energy by
converting β-hydroxybutyrate
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to acetoacetate back to 2 acetyl-CoA

(a reversal of ketogenesis):
 then complete catabolism of 2 acetyl-
CoA in yield 20 ATP in the citric acid cycle
 an additional NADH (2.5 ATP) is produced
by the oxidation of β-hydroxybutyrate to
acetoacetate
 activation of acetoacetate (binding to
Coenzyme-A by succinyl-CoA) requires
one ATP, thereby yielding 21.5 ATPs
 Cont. …12.1 Fatty Acids Biosynthesis

Fatty Acids Biosynthesis:

 fatty acid biosynthesis occur in the cytoplasm of most human and
animal cells, and the liver is the major site for this process
 fatty acids biosynthesis occur when the diet is low in fat or high in
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carbohydrate or protein
 most fatty acids are synthesized from excess dietary carbohydrate
(high acetyl-CoA)
 a large quantity of NADPH is needed for this process and is provided
by the pentose phosphate pathway
 when mitochondrial citrate levels is high (energy requirements are
low), citrate transported to the cytoplasm → cleaved back to acetyl-
CoA and oxaloacetate (catalyzed by citrate lyase)
 acetyl-CoA is then used in fatty acid biosynthesis
 Cont. …12.1 Fatty Acids Biosynthesis

Cont. …Fatty Acids Biosynthesis:

 fatty acid biosynthesis is similar to the reverse of β-oxidation with
the following differences:
 location: occurs in cytoplasm instead of mitochondria
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 enzymes: are significantly different from those in β-oxidation

 thioester linkage: intermediate acyl groups are linked through a

thioester linkage to acyl carrier protein (ACP)

 electron carriers: consumes NADPH (not NADH)

 fatty acid biosynthesis starts with

carboxylation of acetyl-CoA to
form malonyl-CoA, then binds to
ACP forming malonyl-ACP
 Cont. …12.1 Fatty Acids Biosynthesis
Cont. …Fatty Acids Biosynthesis:
 ACC: (acetyl-CoA carboxylase) catalyzes
the carboxylation of acetyl-CoA to
form malonyl-CoA , an irreversible
reaction and uses biotin cofactor
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 ACC: is highly regulated by glucagon

(in hypoglycemia) via allosteric + insulin
phosphorylation (inactivation), and
+ glucagon
insulin (in hyperglycemia) activates ACC
by dephosphorylation (polymerization)
 the remaining reactions of fatty acid
synthesis are catalyzed by the
 AMPK: (AMP Kinase) catalyzes the
multienzyme complex fatty acid
synthetase (FAS) inactivation of ACC
 Cont. …12.1 Fatty Acids Biosynthesis

Cont. …Fatty Acids Biosynthesis:

1. Condensation: acetyl-ACP + malonyl-ACP → acetoacetyl-ACP

+ →
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2. Reduction: of carbonyl group to hydroxyl using NADPH, forming β-

hydroxybutyryl-ACP
3. Dehydration: to crotonyl-ACP
4. Reduction: to butyryl-ACP (4 carbon) , using another NADPH
 then the cycle is repeated with adding 2 carbons acetyl-ACP each
turn till complete fatty acid is formed. for example the synthesis of
palmitic acid (16:0) is repeated 7 times
 acetyl-ACP (2C)

Reduction

acetoacetyl-ACP

Dehydration
β-hydroxybutyryl-ACP

Reduction

butyryl-ACP (4C)
 Cont. …12.1 Fatty Acids Biosynthesis

Regulation of Fatty Acids Metabolism:

 metabolism of fatty acids (the major energy source in animals)
is carefully regulated
 long-term regulation: is the alterations in gene expression of the
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enzyme synthesis
 in short-term regulation: (measured in minutes) the activities of key
regulatory enzymes are modified by allosteric regulators covalent
modification (phosphorylation/dephosphorylation), triggered by hormones
 in the liver in hyperglycemia when insulin/glucagon ratio is high,
malonyl-CoA levels rise and cause the inhibition of β-oxidation
 long-chain fatty acyl-CoA (palmitoyl-CoA) inhibit ACC (acetyl-CoA carboxylase)
by depolymerization
 Cont. …12.1 Fatty Acids Biosynthesis

Cont. …Regulation of Fatty Acids Metabolism:

 when citrate level is high (when energy level is high), citrate is

transferred from inside mitochondria to the cytoplasm, cleaved to
oxaloacetate and acetyl-CoA (catalyzed by citrate lyase), and the
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acetyl-CoA is used for fatty acid synthesis

 when the cell needs more energy, fatty acids are transported into
the mitochondria as acylcarnitine, then acyl-CoA is degraded to
acetyl-CoA via β-oxidation
 glucagon, epinephrine, citrate, malonyl-CoA, and palmitoyl-CoA are
important regulators of fatty acid metabolism
 glucagon (secreted in response to hypoglycemia) activate fatty acid
degradation, possibly by activating AMP Kinase (AMPK)
 Cont. …12.1 Fatty Acids Biosynthesis

Cont. …Regulation of Fatty Acids Metabolism:

 insulin (secreted in response to hyperglycemia) activate fatty acid

synthesis by activating ACC (acetyl-CoA carboxylase)
 high citrate (when energy is sufficient) also activates ACC
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β-oxidation
(mitochondria)
 Cont. …12.1 Fatty Acids Biosynthesis

Cont. …Regulation of Fatty Acids Metabolism:

 AMP: level is a sensitive indicator of cellular energy status, and rise
in response to stress such as nutrient deprivation, hypoxia, heat
shock, and prolonged exercise, that deplete cellular ATP
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 AMPK (AMP kinase): inactivates ACC (acetyl-CoA carboxylase)

 as AMP/ATP ratio rise (low energy), AMP activates AMPK
 once activated, AMPK switches off anabolic pathways (such as fatty
acid and triacylglycerol synthesis) by phosphorylating ACC , and
switches on catabolic pathways (such as β-oxidation)
 AMPK (high AMP): shifts metabolism from energy-consuming
processes to energy-generating processes
 Cont. …12.1 Fatty Acids Biosynthesis

Cont. …Regulation of Fatty Acids Metabolism:

 the main role of activated AMPK in lipid metabolism is to increase
fatty acid uptake into cells and their transport into mitochondria to
be degraded by β-oxidation)
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 Cont. …12. Lipid Metabolism
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