Biochemistry of Metabolism

Metabolismo de las Grasas

Las grasas representan diariamente casi la mitad

de los sustratos para la oxidación

Los principales componentes de la grasa de la dieta

y de los depósitos son los triglicéridos.

Estos consisten básicamente en ácidos grasos

saturados y monoinsaturados esterificados con
glicerol (eg ácidos palmitico, oleico)

Ácidos grasos esenciales, poliinsaturados (eg.

linoleico, linolenico y araquidonico) son
necesarios como precursores de ciertos fosfolipodos
y glucolipidos de membrana

Otro componente de la grasa es la molécula

esteroide colesterol.
 Metabolismo de las Grasas

Grasa excesiva -----Arterosclerosis

La formación de placas cargadas de lípidos
en las paredes arteriales---trombosis
La ingesta de grasa :

No supere el 30% de las calorías totales

La grasa saturada (grasa animal) no supere

10% de las calorías totales

La ingesta de colesterol debe ser inferior a

600 mg/día (reducirse de menos de 300 mg/día
si el colesterol plasmático esta elevado
 Ayuno
El hígado aporte glucosa a la circulación
mediante aumento de la glucogénesis

12 Horas depósitos hepáticos de glucogeno

casi vacíos
Estimulación de gluconeogenesis

2 dias
Degradación diariamente de 75 -100 g proteína
Muscular aumentación de nitrógeno en orina
La gluconeogenesis se mantiene también gracias
al aporte de 15-20g diarios de glicerol,liberados
durante la lipólisis acelerada de los triglicéridos
en el tejido adiposa.
Fatty Acids as Stored Energy
• Fatty acids are the body’s principal
form of stored energy
• Carbon almost completely reduced as
CH2
• Very closely packed in storage
tissues - not hydrated as sugars are
 Fat Storage

• Mainly as triacylglycerols
(triglycerides) in adipose cells
• Constitute 84% of stored energy
• Protein - 15%
• Carbohydrate (glucose or glycogen) - <1%
 Some Facts About Metabolism
 Physiological Mechanisms: Hormones

 Glycogen:
A polysaccharide often referred to as animal starch;
stored in liver and muscle; constitutes the short -term
store of nutrients.
 Insulin:
A pancreatic hormone that facilitates entry of glucose
and amino acids into the cell, conversion of glucose
into glycogen, and transport of fats into adipose tissue.
 Glucagon:
A pancreatic hormone that promotes the conversion of
liver glycogen into glucose.
 Some Facts About Metabolism
 Nutrients

 Glycerol:
 A substance derived from the breakdown of
triglycerides, along with fatty acids; can be converted
by the liver into glucose.

 Fatty acid:
 A substance derived from the breakdown of
triglycerides, along with glycerol; can be metabolized by
most cells of the body except for the brain.
Copyright © 2004 Allyn and Bacon
Release of Fatty Acids from
Triacylglycerols
O
CH 2OC-R1 CH 2OH
O Lipases
CHOC-R 2 CHOH

O
CH 2OC-R3 CH 2OH
Triacylglycerol Glycerol
+
O O O
HOC-R 1 HOC-R 2 HOC-R 3
 Hormone
(Adrenalin, Glucagon, ACTH)
Lipolysis
Receptor

Activates

Adenylyl
Cyclase
ATP c-AMP

Insulin Activates lipase

blocks this
step
Triacylglycerols Glycerol +
Fatty acids Blood

Adipose Cell
Control of triacylglycerol lipase activity in adipose cells
by a cyclic AMP-mediated cascade system.
 Adenylyl cyclase Phosphodiesterase

ATP c-AMP AMP

Enhanced by glucagon Enhanced by insulin

Inactive Kinase Activated Kinase

P

Inactive Lipase Activated Lipase

(Hormone-sensitive
Phosphatase Lipase)

Insulin favors formation

of the inactive lipase Triacyl- Glycerol +
glycerol Fatty Acids
 Acylglycerol Lipases
Diacylglycerol (DAG)
Triacylglycerol
Lipase

OH
Triacylglycerol (TAG) Diacylglycerol
Lipase

OH OH
Monoacylglycerol
Lipase
OH
Monoacylglycerol
OH OH (MAG)

Glycerol
Membrane Transport of
Fatty Acyl CoA Esters
+ O
RCOSCoA + (CH3)3 N
O-
OH
Carnitine Carnitine
Carnitine
acyltransferase II acyltransferase I
(matrix side of (outer part of mitochondrial
inner mitochondrial inner membrane)
membrane)

+ O
(CH3)3 N
O-
O2CR

O-Acylcarnitine

Transported across inner mitochondrial

membrane by translocase
The carnitine cycle, for transport of fatty acyl-CoAs into
mitochondria.
 Beta Oxidation
• Cleavage of fatty acids to acetate in
tissues
• Occurs in mitochondria

[O] [O] [O] [O] [O] [O] [O] [O]

CO2H
9 CH3COSCoA
 Steps in Beta Oxidation
• Fatty Acid Activation by Esterification
with CoASH
• Membrane Transport of Fatty Acyl CoA
Esters
• Carbon Backbone Reaction Sequence
• Dehydrogenation
• Hydration
• Dehydrogenation
• Carbon-Carbon Cleavage (Thiolase Reaction)
 Fatty Acid Activation by
Esterification with CoASH
AcylCoA
CoASH + RCO2H + ATP RCOSCoA + AMP + PPi
Synthetase Pyrophos-
phatase
Occurs in outer mitochondrial
membrane for long chain fatty acids 2 Pi
G0’(KJ/mole)
ATP AMP + PPi -32.3
CoASH + RCO2H RCOSCoA +31.5
PPi 2 Pi -33.6
-34.4
Overview of the fatty acid oxidation pathway.
 Beta Oxidation
Reaction Sequence
Acyl CoA
H H Dehydrogenase H
R-CH2-C-C-COSCoA R-CH2 -C=C-COSCoA
H H FAD FADH2 H trans- 2 -enoyl CoA

H2 O
Enoyl CoA Hydratase
-Ketoacyl CoA L--Hydroxyacyl
CoA H H
H Dehydrogenase
R-CH2 -C-C-COSCoA R-CH2 -C-C-COSCoA

OH NADH NAD + HO H L--Hydroxyacyl CoA

+ H+
CoASH
Thiolase
Occurs in Mitochondria
R-CH2-C-SCoA CH 3 -C-SCoA
+
O O

Repeat Sequence
 Complete Beta Oxidation
of Palmitoyl CoA

CH3CH2--CH2CH2--CH2CH2--CH2CH2--CH2CH2--CH2CH2--CH2CH2--CH2COSCoA

7 Cycles

+
8 CH3COSCoA + 7 FADH2 + 7 NADH + 7 H
 Energetics of Complete
Oxidation of Fatty Acids
High Energy Phosphate
Bonds Generated

Palmitic Acid Palmitoyl CoA -2

TCA Cycle
CH3COSCoA CO2 + H2O 108

Net 106

106 High Energy Phosphate Bonds G0’ = 3,233 KJ/Mole

For Palmitic Acid CO2: Efficiency
G0’ = - 9,790 KJ/Mole of -Oxidation = 33%
 Beta Oxidation of Odd
Carbon Fatty Acids
CH3CH2CH2--CH2CH2--CH2CH2--CH2CH2--CH2CH2--CH2COSCoA

5 Cycles

5 CH3COSCoA + CH3CH2COSCoA Propionyl CoA

Propionyl CoA Carboxylase
TCA Cycle ATP/CO2

CO2H CO2 H
Mutase Epimerase

HO 2CCH 2CH2COSCoA
CH 3 -C-H H-C-CH 3

Succinyl CoA Vit. B12 COSCoA COSCoA

L-Methylmalonyl D-Methylmalonyl
CoA CoA
 Beta Oxidation of
Unsaturated Fatty Acids
H H
CH 3(CH2)7-C=C-CH 2(CH2)6COSCoA
Oleoyl CoA
Beta Oxidation
(3 Cycles)

H
H H Isomerase

CH 3(CH2)7-C=C-CH 2COSCoA CH 3(CH2)7-CH2-C=C-COSCoA

trans- 2 H
cis- 3

Continuation of Beta Oxidation

Oxidation pathway for
polyunsaturated fatty
acids.
 Refsum’s Disease
• A rare inherited disorder in which
phytanic acid accumulates in tissues:
• Possibly due to defect or deficiency of the -
hydroxylase
• Nerve and retinal damage
• Spastic movement
• Bone and skin changes
• Treatment: Avoidance of chlorophyll-
containing foods, including meat from plant-
eating animals
 Ketogenesis (Ketosis):
Formation of Ketone Bodies
Thiolase
2 CH3COSCoA CH3COCH2COSCoA CH3COSCoA
Acetoacetyl CoA
HMG CoA
Synthase

Several
Cholesterol steps OH
(in cytosol)
HO2C-CH2-C-CH2COSCoA
CH3

Ketogenesis -Hydroxy--methylglutaryl CoA

(HMG CoA)
 Ketogenesis: Formation
of Ketone Bodies (Cont’d.)
OH
HMG CoA
HO2C-CH2-C-CH2COSCoA lyase
CH3COCH2CO2H
CH3 - CH3COSCoA Acetoacetic Acid
HMG CoA NADH + H+
Dehydrogenase - CO2
NAD+

OH
CH3COCH3
CH3CHCH2CO2H
Acetone
-Hydroxybutyrate (volatile)

Ketone bodies are important sources

of energy, especially in starvation
 Regulation of Ketogenesis

J. Denis McGarry

• Activity of ketogenic enzymes is not regulated.

• Ketogenesis rate in isolated mitochondria is unchanged in
diabetic animals.
 Ketone Bodies As
Energy Sources
-Hydroxybutyrate Acetoacetate Succinyl CoA

-Ketoacyl CoA
transferase

Thiolase
Acetoacetyl CoA Succinic acid
2 Acetyl CoA

TCA Cycle
Ketones in Diabetes Mellitus
Ketogenic
Amino Acids Fatty
Glycolysis Acids
Glucose Oxaloacetate
Gluconeogenesis Acetyl CoA

TCA Ketone
Glucogenic Cycle Bodies
Amino Acids
Citrate

Glucose in cells Gluconeogenesis Oxaloacetate

Fatty Acid breakdown Acetoacetyl CoA Ketone Bodies

 Fatty Acid Synthesis vs.
Degradation
Synthesis Degradation

Intermediates Linked to SH in Linked to CoASH

Proteins
(Acyl Carrier Proteins)

Site Cytosol Mitochondria

Enzymes Components of Separate Polypeptides

Single Peptide

Redox +
NADP / NADPH
+
NAD / NADH
Coenzymes
 Fatty Acid Biosynthesis

• Occurs in cytosol
• Starts with acetyl CoA
• Problem:
» Most acetyl CoA produced in mitochondria
» Acetyl CoA unable to traverse mitochondrial
membrane
 Cytosol Mitochondria
Fatty Acids
Pyruvate Beta
Oxidation
Dehydrogenase
Glucose Pyruvate Pyruvate Acetyl CoA

Fatty Acids
ATP-Citrate Oxalo-
Lyase Citrate acetate
Acetyl CoA Citrate

Amino acids

Mitochondrial
membrane
Acetyl-CoA as a key intermediate between fat
and carbohydrate metabolism.
 Fatty Acid Biosynthesis:
Formation of Malonyl CoA
Acetyl CoA
Carboxylase Malonyl CoA
CH3COSCoA + ATP + HCO3- -O CCH COSCoA
2 2

+ ADP + Pi + H+
• Committed step in fatty acid synthesis
• Reaction is irreversible
• Regulation of acetyl CoA carboxylase activity:
by palmitoyl CoA
by citrate
• Malonyl CoA inhibits carnitine acyl transferase I
• Blocks beta oxidation
 Fatty Acid Biosynthesis:
Role of Acyl Carrier Proteins
Acetyl
Transferase
CH3COSCoA CH3CO-S-ACP
Acetyl ACP
Malonyl
Transferase
-O CCH COSCoA -O CCH CO-S-ACP
2 2 2 2
Malonyl ACP

ACP = Acyl carrier protein

 Fatty Acid Biosynthesis:
Formation of Acetoacetyl ACP

CH3CO-S-ACP + -O2CCH2CO-S-ACP
-Ketoacyl ACP
Synthetase

CH3COCH2CO-S-ACP + CO2
Acetoacetyl ACP
 Fatty Acid Biosynthesis:
Formation of Butyryl ACP
-Ketoacyl ACP OH
reductase
CH3COCH2CO-S-ACP CH3CCH2CO-S-ACP
Acetoacetyl ACP NADPH NADP+ H
+ H+ -D-Hydroxybutyryl ACP
-Hydroxyacyl ACP
- H2O dehydratase

NADP+ NADPH H
+ H+

CH3CH2CH2CO-S-ACP CH3C=C-CO-S-ACP
2,3-trans-
Butyryl ACP Enoyl ACP Crotonyl ACP
H
reductase
 Fatty Acid Biosynthesis:
Chain Elongation
CH3CH2CH2CO-S-ACP -O CCH CO-S-ACP
+ 2 2

CH3COCH2CH2CH2CO-S-ACP

OH H

CH3CCH2CH2CH2CO-S-ACP CH3C=CCH2CH2CO-S-ACP
H H
 Fatty Acid Biosynthesis:
Chain Elongation (Cont’d)
H NADPH NADP+
+ H+

CH3C=CCH2CH2CO-S-ACP CH3(CH2)3CH2CO-S-ACP
H
5 Cycles

- CH3(CH2)13CH2CO-S-ACP
CH3(CH2)13CH2CO2
Palmitoyl ACP
Palmitate
Further Processing of Fatty
Acids: Elongation
In mitochondria and
- at surface of
CH3(CH2)13CH2CO2 endoplasmic reticulum
Palmitate
CH3COSCoA
Thiolase

CH3(CH2)13CH2COCH2COSCoA
NADH + H+
Dehydrogenase

NAD+ OH L- Configuration

CH3(CH2)13CH2CCH2COSCoA
H
Further Processing of Fatty
Acids: Elongation (Cont’d)
OH
CH3(CH2)13CH2CCH2COSCoA

- H2O H
Hydratase
H
CH3(CH2)13CH2C=CCOSCoA
NADPH + H+
Dehydrogenase
H
NADP+

CH3(CH2)13CH2CH2CH2COSCoA
Stearoyl CoA
 Reactions of Fatty Acid Biosynthesis
Reactions 1 and 2 are “Priming”
Dehydration

Reduction:NADPH

6 additional cycles
Condensation and loss of CO2

(3)

Reduction:NADPH

ENZYME STOPS AT C:16:0

Metabolite Regulation of Fatty
Acid Synthesis and Breakdown
Glucose Citrate
Stimulates
Blocks
Beta
Pyruvate Acetyl CoA Malonyl CoA
Oxidation

Inhibits

Palmitoyl CoA
 Hormonal Regulation of Fatty
Acid Synthesis and Breakdown

Adenylyl cyclase Phosphodiesterase

ATP cAMP AMP

Stimulates Stimulates

Glucagon Insulin
Inactivates
Activates Protein Kinase lipase

Inhibition of Activates triacyl-

Inactivates ACC by
fatty acid glycerollipase
phosphorylation
synthesis
Acetyl CoA Carboxylase Catalyzes the Committed and Rate
Limiting Step of Fatty Acid Biosynthesis

Allosteric Effectors: citrate (+);

HIGHLY REGULATED palmitoyl CoA (-)
Phosphorylation/Dephosphoryla
tion; Enzyme-P is inactive
Transcription: e.g., high
carbohydrate diets stimulate
 REGULATION OF FATTY ACID BIOSYNTHESIS

Regulatory Agent Effect

PALMITATE BIOSYNTHESIS

Citrate Allosteric activation

Short Term

C16-C18 acyl CoAs Allosteric inhibition

Insulin Stimulation
Glucagon Inhibition
cAMP-mediated phosphorylation Inhibition
Dephosphorylation Stimulation
Long Term

High carbohydrate diet Stimulation by increased enzyme synthesis

Fat-free diet Stimulation by increased enzyme synthesis
High-fat diet Inhibition by decreased enzyme synthesis
Fasting Inhibition by decreased enzyme synthesis
Glucagon Inhibition by decreased enzyme synthesis
Allosteric Regulation

Well-fed
Regulation by Phosphorylation

Lipogenic
Allosteric Regulation

Fasting
Regulation by Induction

Well-fed
Regulation by Induction

Fasting
Lehninger Principles of Biochemistry
 Regulation of Ketogenesis
• Palmitate requires CPT for transport into
mitochondria.
• Octanoate is freely permeable across mitochondrial
membranes.
• octanoylcarnitine is an inhibitor of CPT.

Conclusions?
 Overview

Cells convert carbons from carbohydrates, amino acids, and other

lipids to fatty acids as long as they can be first metabolized to
acetyl-CoA.

Fatty acid synthesis occurs primarily in the liver and lactating

mammary gland, and to a lesser extent in adipose tissue and
kidney. The major intracellular site of synthesis of acetyl-CoA is
the mitochondrion, whereas fatty acid synthesis takes place in the
cytosol; therefore, a pathway for efflux of the acetyl-CoA is
needed. This is achieved via the citrate shuttle :

Cytosol: Mitochondrion:
Glycolysis ---> Pyruvate ---[Transport]---> Pyruvate ---> Acetyl-CoA
Acetyl-CoA ---> Citrate
Citrate <---[Transport]--- Citrate

Citrate ---> Acetyl-CoA

 Mechanism for transfer of acetyl CoA from mitochondria to cytosol for
fatty acid biosynthesis

Å¢¢Å@Ä-f9«1Ü!

CoA +

Figure 9.10
 Fatty Acid Synthase Catalyzes the Biosynthesis of Palmitate

In humans, this enzyme is a dimer that has seven (7)

different activities: ACP = acyl carrier protein(in bacteria), in
mammals it is a domain of the fatty acid synthase (PhP
domain)
OVERVIEW
• Fatty acid synthesis involves the condensation of a “priming”
molecule of acetyl-CoA plus malonyl-CoA.
• The iterative cycling of the enzyme to synthesize a fatty acid
involves reactions similar to the reversal of fatty acid oxidation,
to grow the chain in 2-carbon units:
Acetyl-CoA (C2:0) [+ malonyl-CoA] --->
Butyryl-CoA (C4:0) [+ malonyl-CoA] --->
Hexanoyl-CoA (C6:0) [+ malonyl-CoA] ---> etc.