CHAPTER 8: Lipid Metabolism

Wednesday, 27 March 2019 10:14 AM

DIGESTION AND ABSORPTION OF LIPIDS

Chyme - a thick semi-liquid material made up of small TAG globules, other partially digested food
and gastric secretions (HCl and several enzymes).
- High fat foods remain in the stomach longer than low fat foods.
- Lipid digestion begin in the stomach where gastric lipase hydrolyzes TAG (10%).
- In the small intestine bile is released, triggered by cholecystokinin.
○ Bile acts as emulsifier
○ Pancreatic lipase major enzyme involve which hydrolyzes the ester linkages between
glycerol and FA of TAGs.
○ Hydrolysis of TAG from bile = free fatty acids and monoacylglycerols (MAG) which forms
micelles.
❖ Fatty Acid Micelle - a micelle in which fatty acids and or MAG and some bile are present.
○ Can be absorbed through the (enterocytes) membranes of intestinal walls .
- Within the enterocytes free FA and MAG are repackaged into TAG, then combined with
membrane lipids and proteins to produce a:
○ Chylomicron - a lipoprotein that transports TAG from intestinal cells, via the lymphatic
system, to the bloodstream.
- Enters the lymphatic system through the thoracic duct
- Concentrations usually rise within a 2 hours after a meal, peak in 4-6 hours and
drop rapidly to normal level.
- TAG released from chylomicrons are mediated by lipoprotein lipase which are located in the
lining of blood vessels.
- FA and glycerol from TAG are either:
○ Absorbed by the cells.
○ Broken down to Acetyl-CoA.
○ Stored as lipids.

TRIACYLGLYCEROL STORAGE AND MOBILIZATION

Adipocyte - TAG-storing cell.
Adipose tissue - tissue that contains large number of adipocyte cells.
- Serve as insulator against excessive heat loss
- Provides organs with protection against physical shock.
Epinephrine and Glucagon - hormones that triggers use of TAG in adipose tissue for energy
production.
- Hormonal interactions with adipose cell membrane receptors stimulates cAMP production
from ATP in adipose cell.
- In turn, cAMP activates hormone-sensitive lipase (HSL) through phosphorylation.
○ HSL - lipase for hydrolysis of TAG, prerequisite for FA to enter bloodstream from adipose
cell.
Triacylglycerol mobilization - the hydrolysis of TAG stored in adipose tissue, followed by release in
into the bloodstream of FA and glycerol so produced.
- TAG is the body's major source of stored energy.

GLYCEROL METABOLISM
- 1 TAG completely hydrolyzed = 1 glycerol
- After entering bloodstream, glycerol travels to liver or kidney where 1st step of glycerol
metabolism occurs.
○ Glycerol is converted to dihydroxyacetone phosphate (DHAP).
○ DHAP can be converted to pyruvate, then Acetyl-CoA and finally CO2 or to glucose.
- Glycerol to DHAP is a 2 step ATP consuming process:
1. Phosphorylation of primary hydroxyl group of glycerol.
2. Glycerol's secondary alcohol group (C2) is oxidized to a ketone.

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 2. Glycerol's secondary alcohol group (C2) is oxidized to a ketone.
Glycerol > G3P > DHAP

OXIDATION OF FATTY ACIDS

3 Parts process by which FA is broken down to obtain energy.
1. The FA must be activated by bonding to Coenzyme A.
2. The FA must be transported into the mitochondrial matrix by a shuttle mechanism.
3. The FA must be repeatedly oxidized, cycling through a series of four reactions to produce
acetyl CoA, FADH2, and NADH.

I. Fatty Acid Activation

❖ Outer mitochondrial membrane - FA activation site.
❖ Reactants: FA, Acetyl CoA, and a molecule of ATP
❖ Enzyme: Acyl CoA synthetase
❖ FA is converted to acyl CoA (activated FA-CoA molecule):
○

- Requires 2 phosphate bonds from ATP

- ATP is converted to AMP and 2Pi.
- Acyl: random-length FA carbon chain that is covalently bonded to CoA.
- Acetyl: two-carbon chain covalently bonded to CoA.
II. Fatty Acid Transport
- Acyl CoA is too large to pass through the inner mitochondrial membrane to the matrix, needs a
shuttle mechanism.
- Shuttle mechanism involving carnitine facilitates Acyl CoA which carries it through the
matrix.
- The acyl group is then transferred from carnitine back to a CoA molecule.
III. Reactions of β-Oxidation Pathway
- A repetitive series of four biochemical reactions that degrade acyl CoA to acetyl CoA by
removing two carbon atoms at a time with FADH2, and NADH also being produced.
- 4 reaction sequence:

Step 1: First dehydrogenation.

- Hydrogen atoms are removed from α and β carbons creating a double bond between 2
carbon atoms.
- Oxidizing agent: FAD
- Product: trans-Enoyl CoA
- Enzyme: Acyl CoA Dehydrogenase (stereospecific)

Step 2: Hydration.
- A molecule of water is added to the trans double bond producing a secondary alcohol at
the β-carbon position.
- Enzyme: Enoyl CoA hydrase (stereospecific)
- Product: L-β-Hydroxyacyl CoA

Step 3: Second dehydrogenation.

- Removal of 2 hydrogen atoms converts β-Hydroxy to a keto group.
- Oxidizing agent: NAD+
- Enzyme: β-Hydroxyacyl CoA dehydrogenase (stereospecific)
- Product: β-Ketoacyl CoA

Step 4: Thiolysis.

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 Step 4: Thiolysis.
- The FA carbon chain is broken between the α and β carbons by reaction with coenzyme
A molecule.
- Enzyme: Thiolase
- Product: Acetyl CoA and Acyl CoA with 2 less carbons

- Acyl CoA is recycled through the same four reactions until the entire FA is converted to acetyl
CoA.
- Number of Acetyl CoA molecules produced in β-oxidation is equal to the half of carbon atoms
in FA.
- Number of repetitions of β-oxidation is less than one of the acetyl CoA produced because the
last repetition produces 2 acetyl CoA molecules as C4 units split into C2 units.
Unsaturated Fatty Acids:
- Oxidation through β-oxidation pathway requires 2 additional enzymes aside from those
needed in saturated FAs.
- Additional enzymes: epimerase and cis-trans isomerase.
- Epimerase can change the D form to L form of cis double bonds in naturally
occurring unsaturated FA.
- Cis-trans isomerase produces a trans-(2,3) double bond from a cis-(3,4) double
bond.

ATP PRODUCTION FROM FATTY ACID OXIDATION

- Lipids are 33% more efficient than carbohydrates as energy-storage systems.
- 1g of carbohydrates = 4kcal
- 1g of fat = 9kcal
Calculating net ATP from oxidation of FA:
Lauric Acid = C12 saturated FA
No. of Acetyl CoA (half of the no. of C atoms) 6
No. of β-oxidation repetitions (Acetyl CoA less 1) 5
Activation -2 ATPs
Computation:
β-Oxidation (FADH2 and NADH):

Acetyl-CoA (10 ATPs):

6 Acetyl CoA x 10 = 60 ATPs

TOTAL:

19 C Saturated FA = 19 C Fatty Acyl-CoA

Will be released as propionyl-S-CoA /\/\/\/\/\/\/\/\/\C-S-CoA
/ = will be β-oxidized
No. of β-oxidation 8 (NADH, FADH2)
No. of Acetyl CoA 8
Propionyl-S-CoA to Methyl malonyl-S-CoA -1 ATP
Activation -2 ATP
Computation:
Succinyl-S-CoA:
1 GTP = 1 ATP

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 1 GTP = 1 ATP
1 FADH2 = 1.5 ATPs
1 NADH = 2.5 ATPs
= 5 ATPs
8 Repetitions of β-oxidation:
8 NADH x 2.5 = 20 ATPs
8 FADH2 x 1.5 = 12 ATP
8 Acetyl CoA x 10 = 80 ATPs

TOTAL:

Fuel Uses:
1. Skeletal muscles uses glucose in active state, fatty acids in resting state.
2. Cardiac muscle depend first on FA and secondarily on ketone bodies, glucose, and
lactate.
3. Liver prefers FA.
4. Brain is maintained by glucose and ketone bodies. FA cannot cross blood-brain barrier.

KETONE BODIES AND KETOGENESIS

- Oxaloacetate (OAA) concentration depends on pyruvate produced from glycolysis.
- Pyruvate can be converter to OAA by pyruvate carboxylase.
Body conditions that affect lipid-carbohydrate balance:
• Dietary intake high in fat and low in carbohydrates (keto diet).
• Diabetic conditions.
• Prolonged fasting conditions.
Ketone body - one of three substances produced from acetyl CoA when an excess of acetyl CoA from
FA degradation accumulates because of TAG-carbohydrate metabolic imbalances.
3 Ketone bodies
❖ Acetoacetate (acid)
❖ β-hydroxybutyrate (acid)
❖ Acetone
- Reduction of ketone group in acetoacetate produces to a secondary alcohol β-
hydroxybutyrate.
- Decarboxylation of acetoacetate produces acetone (volatile).
Ketogenesis
- Is the metabolic pathway by which ketone bodies are synthesized from acetyl CoA.
- Liver mitochondria is the primary site for ketogenesis.
- Acetoacetate and β-hydroxybutyrate are produced in the liver.
- Acetone is produced in the bloodstream, not a product of metabolic pathway
ketogenesis.
- Acetone is excreted by exhalation, can be smelled from breath of diabetic.
- Ketogenesis process:
Step 1: First condensation.
- 2 Acetyl CoA combines via condensation to produce acetoacetyl CoA.

Step 2: Second condensation.

- Acetoacetyl CoA reacts with another acetyl CoA and water to produce 3-
hydroxy-3-methyglutaryl (HMG-CoA) and CoA-SH.

Step 3: Chain cleavage.

- HMG-CoA is cleaved to acetyl CoA and acetoacetate.

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 - HMG-CoA is cleaved to acetyl CoA and acetoacetate.

Step 4: Hydrogenation.
- Acetoacetate is reduced to β-hydroxybutyrate.
- Reducing agent: NADH

Energy Production from Acetoacetae

- Liver lacks the enzyme (β-ketoacyl transferase) to generate acetyl CoA from ketone
bodies.
- Ketone bodies are water soluble and doesn’t need a protein transport system.
- For acetoacetate to be used as a fuel, it must be activated first by:
- Transfer of CoA group from succinyl CoA.
Ketosis
- Body condition in which high level of ketone bodies are present in blood and urine.
Contributors:
Ketonemia - 20 mg/100 mL of ketone.
Ketonuria - 70 mg/100 mL
- Ketoacidosis: serious ketosis problem and can occur in people with Type 1 diabetes.

BIOSYNTHESIS OF FATTY ACIDS: LIPOGENESIS

Lipogenesis
- The metabolic pathway by which fatty acids are synthesized from acetyl-CoA
- Occurs in cell cytosol; degradation of FA occurs in mitochondrial matrix.
- Enzymes are collected in to a complex called fatty acid synthase.
- Lipogenesis intermediates are bonded to acyl carrier protein (ACP). While CoA is the
degradation intermediate.
- FA synthesis is dependent on NADPH, FA degradation is dependent FAD and NAD+.
- Occurs anytime dietary intake provides more nutrients than are needed for energy
requirements.
- Primary lipogenesis sites:
• Liver
• Adipose tissue
• Mammary glands
Citrate-Malate Shuttle System
- Inner mitochondrial membrane is not penetrable to acetyl CoA, needs shuttle system
• Mitochondrial acetyl-CoA reacts with OAA to produce citrate, which is transported
through the inner mitochondrial membrane by a citrate transporter.
• Once in the cytosol, citrate undergoes reverse reaction to regenerate acetyl CoA and
OAA with ATP involved.
• Acetyl CoA becomes fuel.
• OAA becomes malate and reenters the mitochondrial matrix through malate transporter
and converted back to OAA to shuttle another Acetyl CoA and repeat the process.
ACP Complex Formation
- Involved in ACP structure are 2-ethanethiol and pantothenic acid components present in CoA-
SH.
- 2 ACP complexes are needed to start lipogenesis process:
• Acetyl ACP (C2-ACP)
- Produced by reaction of acetyl-CoA with ACP molecule.
• Malonyl ACP (C3-ACP)
- 2 steps:
1. Carboxylation reaction with ATP involvement:

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 2. Malonyl CoA produced reacts with ACP to produce malonyl ACP.
Chain Elongation
Step 1: Condensation.
- Acetyl ACP and malonyl ACP condense together to form acetoacetyl ACP.
Step 2: First hydrogenation.
- Keto group of acetoacetyl complex, which involves β-carbon atm, is reduced to the
corresponding alcohol by NADPH.
Step 3: Dehydration.
- Alcohol produced in step 2 is dehydrated to introduce a double bond into the molecule
between α and β carbons.
Step 4: Second hydrogenation.
- The double bond is converted to a single bond through hydrogenation, NADPH is the
reducing agent.
- Elongation stops upon formation of C16 acyl group (palmitic acid).

Unsaturated Fatty Acid Synthesis

- Production of unsaturated fatty acids requires molecular oxygen.
Relationships Between Lipogenesis and Citric Acid Intermediates
1. The citric acid intermediates involve C4 diacids, and the lipogenesis intermediates involve C4
monoacids.
2. The order in which the various acid derivative types are encountered in the lipogenesis is the
reverse in which they are encountered in the citric acid cycle.

FATES OF FATTY-ACID-GENERATED ACETYL COA

1. Can be further processed through the common metabolic pathway (CAC, ETC and oxidative
phosphorylation) to get ATP.
2. Can become ketone bodies.
3. Can be stored as TAG.
4. Can be used to synthesize lipids.
5. Cannot be used for net synthesis of glucose.

RELATIONSHIP BETWEEN LIPID AND CARBOHYDRATE METABOLISM

- Acetyl CoA is the primary link between lipid and carbohydrate metabolism.
- Acetyl CoA is the degradation product for glucose, glycerol, and FA.
- Acetyl CoA is the starting material for biosynthesis of FA, cholesterol, and ketone bodies.

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