 Contents :

 Lipolysis
 Beta oxidation
 Alpha oxidation
 Omega oxidation

By: Savath Sovannak

 By: Savath Sovannak

 Definition : Lipolysis is the hydrolysis of triacylglycerols into glycerol and fatty acids
 Location : cytosol of adipose tissue cells
 Step : Lipolysis Is earned out by a number of lipase enzymes, which are present in adipose tissue.
 Adipose triglyceride lipase (ATGL) : cleave TAG to diacylglycerol (DAG) and Free fatty acid
 Hormone sensitive lipase (HSL) : Cleave DAG to MAG and Free fatty acid (FFA)
 Monoacylglycerol lipase (MAG lipase): Cleave MAG to Glycerol and FFA
 Regulation of Lipase enzymes

 Fat droplets are coated by a protein (perilipin) bind with comparative gene identification
58 (CGI-58)  limits access of ATGL and HSL
 Phosphorylation of perilipin 
o Release CGI-58 from perilipin  activate ATGL
o allows translocation and binding of phosphorylated HSL to the droplet
  Epinephrine, norepinephrine and glucagon  (+)
adenylate cyclase enzyme  ↑cAMP  (+) PKA
 phosphorylate perilipin and HSL  (+) lipolysis
 Insulin  (+) phosphodiesterase 3b ↓cAMP and
PKA  (-) lipolysis
 Fate of glycerol
 Glycerol released during TAG degradation cannot
be metabolized by adipocytes because they lack
glycerol kinase transported through blood to
liver convert to glycerol 3-phosphate by glycerol
kinase  can be used to
o form TAG in the liver
o converted to DHAP by reversal of the
glycerol 3-phosphate dehydrogenase
 Fate of FFA
 FFA move through the cell membrane of the
adipocyte and bind to serum albumin
transported to tissues such as muscle, liver,
enter cells, get activated to their CoA derivatives,
and are oxidized for energy in mitochondria

 Regardless of their levels, plasma FFA cannot be used for fuel by red blood cells (RBC), which have
no mitochondria.
 The brain does not use fatty acids for energy to any appreciable extent, but the reasons are less
clear

By: Savath Sovannak

 Contents :
 Introduction
 FA transport into mitochondria
 Beta-oxidation of even saturated short-LCFA
 Beta-oxidation energy yield
 Beta-oxidation of odd chain FA
 Unsaturated FA oxidation
 Peroxisomal beta oxidation

By: Savath Sovannak

 There are 3 oxidation of FA :
 Beta-oxidation
 Alpha-oxidation
 Omega-oxidation
 Metabolism site:
 Beta oxidation : in either mitochondria or peroxisomal of several tissues, including liver,
muscle, and adipose tissue
o Note : RBC and brain are not able to utilize FA
 Alpha-oxidation : in peroxisomal
 Omega-oxidation : in smooth endoplasmic reticulum (SER)
 Beta oxidation : a catabolic process in which a FA chain is cleaved (oxidized) at the beta
carbon

By: Savath Sovannak

 SCFA and MCFA : diffuse freely into mitochondria. Once inside the mitochondria, they are activated to
their CoA derivatives by matrix enzymes and are oxidized
 LCFA :
 After a LCFA enters a cell converted in cytosol to LCF acyl CoA by long-chain fatty acyl CoA
synthetase (thiokinase), an enzyme of the outer mitochondrial membrane
 Beta-oxidation occurs in the mitochondrial matrix  LCFA-CoA must be transported across the
mitochondrial inner membrane, which is impermeable to CoA  need specialized carrier :
Carnitine. The transport process is called carnitine shuttle
 Carnitine shuttle :
o Structure of carnitine :
• Carnitine is 𝛽-hydroxy-𝛾-trimethyl ammonium butyric acid
• Source :
 can be obtained from the diet, where it is found primarily in meat products
 It can also be synthesized from the amino acids lysine and methionine by an
enzymatic pathway :
 found in the liver and kidneys but not in skeletal or cardiac muscle
 Skeletal muscle contains 97 % of all carnitine in the body (dependent on
uptake of carnitine provided by endogenous synthesis or the diet and
distributed by the blood)

By: Savath Sovannak

 o Translocation step :
• Acyl group is transferred from CoA to
carnitine by carnitine
palmitoyltransferase I (CPT-I = carnitine
acyltransferase I) form acylcarnitine
and free CoA
• Acylcarnitine is transported into the
mitochondrial matrix in exchange for free
carnitine by carnitine-acylcarnitine
translocase (CACT)
• Carnitine palmitoyltransferase 2 (CPT-II,
or CAT-II)  transfer acyl group from
carnitine to CoA  form LCF acyl-CoA
(acyl-CoA in short) and free carnitine

o Carnitine shuttle inhibitors :

• Malonyl CoA inhibits CPT-I  prevent LCFA-CoA enter matrix
  when fatty acid synthesis is occurring in the cytosol (as indicated by the presence of
malonyl CoA), the newly made palmitate cannot be transferred into mitochondria and
degraded
 Muscle tissue, although it does not synthesize fatty acids, contains the mitochondrial isozyme
of ACC (ACC2), allowing regulation of Beta-oxidation. The liver contains both isozymes.

By: Savath Sovannak

 In Short-LCFA with saturated even chain  It consists of a sequence of four reactions
1. First Oxidation (Dehydrogenation)
 Acyl-CoA convert by (short, medium, Long ) chain fatty acyl CoA
dehydrogenase to :
o Trans-2-Enoyl CoA
o FADH2
2. Hydration
 Trans-2-Enoyl CoA convert by 2,3-enoyl CoA hydratase to
o 3-hydroxyacyl CoA
3. Second Oxidation (dehydrogenation)
 3-hydroxyacl CoA convert by 3-hydroxyacyl CoA dehydrogenase to
o 3-ketoacyl CoA
o NADH + H+
4. Thiolytic cleavage
 3-ketoacyl CoA convert by 3-ketoacyl CoA thiolase to
o Acetyl CoA
o Fatty acyl CoA (shorten by 2C atom)
 New acyl CoA can enter beta oxidation again until only 2 molecules of acetyl-CoA are
left over
 These four steps are repeated for saturated fatty acids of even-numbered carbon
chains (n/2) -1 times
 n is the number of carbons
 Each cycle producing :
o One acetyl CoA except the final cycle produce 2 acetyl CoA
o One FADH2 and one NADH + H+

By: Savath Sovannak

 The energy yield from fatty acid 𝛽-
oxidation is high
 Ex : Oxidation of a palmitoyl CoA (16
carbon)  form
 8 acetyl CoA
 7 (FADH2 and NADH + H+)
 1 Acetyl CoA  Krebs cycle  ATP, 3
NADH, CO2, FADH2
 So 8 Acetyl  form 8 ATP, 24 NADH,
8 FADH2, 8 CO2
 Total : 1 palmitoyl CoA  form
 8 ATP - 2 ATP (use to activate FA) = 6
 31 NADH
 15 FADH2
 េបើ 1 NADH form 2.5 ATP and 1FADH2
form 1.5 ATP
 31 NADH = 31 (2.5) = 77.5
 15 FADH2 = 15 (1.5) = 22.5
 Total : 77.5 + 22.5 + 6 = 106 ATP
 េបើ 1 NADH form 3 ATP and 1FADH2 form
2 ATP
 31 NADH = 31 (3) = 93
 15 FADH2 = 15 (2) = 30
 Total : 93 + 30 + 6 = 129 ATP
By: Savath Sovannak
 This process proceeds by the same reaction steps as that of fatty acids with
an even number of carbons, until the final three carbons are reached
 Final cycle of fatty acid oxidation :
 Degradation of FA with saturated even chain  form 2 acetyl-CoA (out
of a 4 carbon fragment)
 Degradation of FA with saturated odd chain  form 1 acetyl-CoA and 1
propionyl-CoA (out of a 5 carbon fragment)
 Propionyl CoA is metabolized by a three step pathway
1. D-Methylmalonyl CoA synthesis :
 Propionyl-CoA is carboxylated, forming D-methylmalonyl-CoA by
propionyl-CoA carboxylase that require biotin and ATP as
coenzyme
o Biotin bind to propionyl-CoA carboxylase at amino group of
lysine residue of enzyme  form Enzyme-Biotin
o Hydrolysis ATP  form Enzyme-Biotin- CO2 (HCO3  CO2)
o Enzyme-Biotin- CO2  transfer CO2 to propionyl -CoA 
form D-methylmalonyl-CoA
2. L-Methylmalonyl CoA formation : D-isomer is converted to the L -form
by enzyme methylmalonyl CoA racemase.
3. Succinyl CoA synthesis :
 L-Methylmalonyl -CoA is converted to succinyl-CoA by enzyme
methylmalonyl CoA mutase that require vitamin B12 as coenzyme

By: Savath Sovannak

  The oxidation of unsaturated fatty acids generates
intermediates that cannot serve as substrates for acyl
CoA dehydrogenase and 2,3-enoyl CoA hydratase 
additional enzymes are required​
 The additional enzyme is depend on position of
double bond
 Oxidation of a double bond at an odd-
numbered carbon  requires 3,2-enoyl CoA
isomerase
 Oxidation of a double bond at an even-
numbered carbon  requires NADPH-
dependent 2,4-dienoyl CoA reductase
 β-oxidation of mono-unsaturated fatty acids :
 Oleic acid (18: Δ9) is the example of mono-unsaturated FA that double bound is at odd-
numbered carbon
 In the first step of oxidation, oleate is converted to oleoyl CoA and, like the saturated
fatty acids, enters the mitochondrial matrix via the carnitine shuttle.
 Oleoyl-CoA then undergoes three β-oxidation cycles to yield three molecules of acetyl-
CoA, 3 NADH and 3 FADH2 and cis-3 (or Δ3)-dodecenoyl-CoA
 Cis-3-dodecenoyl-CoA cannot serve as a substrate for acyl CoA dehydrogenase and also
for enoyl-CoA hydratase, which acts only on trans double bonds  3,2-enoyl CoA
isomerase isomerizes the cis-3-enoyl-CoA to the trans-2-enoyl-CoA can convert by
enoyl CoA hydratase into β (or 3)-hydroxyacyl CoA (trans-Δ2-dodecenoyl CoA)  acted
upon by the remaining enzymes of β oxidation  form acetyl CoA, FADH2, NADH and
remain 10C saturated acyl CoA (decanoyl CoA)
 10C saturated acyl CoA (decanoyl CoA) undergoes four more passes through the β-
oxidation pathway to yield five more molecules of acetyl CoA, 4(FADH2 and NADH + H+)
By: Savath Sovannak
 β-oxidation of poly-unsaturated fatty acids
 Linoleic acid (18:2 Δ9,12) is the example of poly-
unsaturated FA that double bound is at even-numbered
carbon
 In the first step of oxidation, Linoleic acid is converted to
linoleoyl CoA then enters the mitochondrial matrix via
the carnitine shuttle.
 Linoleoyl CoA undergoes three β-oxidation cycles to yield
three molecules of acetyl CoA , 3(FADH2 and NADH + H+),
and 12 C unsaturated fatty acyl CoA with 2 double bond
at cis-Δ3,cis-Δ6
 This intermediate cannot serve as a substrate for acyl
CoA dehydrogenase and also for enoyl-CoA hydratase 
3,2-enoyl CoA isomerase convert Fatty acyl-CoA cis-
Δ3,cis-Δ6 to Fatty acyl-CoA trans-Δ2,cis-Δ6 can convert
by enoyl CoA hydratase and other remaining enzymes of
β oxidation  form acetyl CoA, FADH2, NADH and 10C
Fatty acyl-CoA cis-Δ4
 Fatty acyl-CoA cis-Δ4 is substrate for acyl CoA
dehydrogenase  form Fatty acyl-CoA trans-Δ2, cis-Δ4
 Fatty acyl-CoA trans-Δ2, cis-Δ4 is not substrate for enoyl-
CoA hydratase  need NADPH dependent 2,4-dienoyl-
CoA reductase  convert Fatty acyl-CoA trans-Δ2, cis-Δ4
to NADP+ and fatty acyl-CoA trans-Δ3
 Then enoyl-CoA isomerase convert fatty acyl-CoA trans-Δ3 to
fatty acyl-CoA trans-Δ2  can enter the β-oxidation pathway

By: Savath Sovannak

 VLCFAs undergo 𝛽-oxidation in peroxisomes
 First VLCFA is activated in cytosol like LCFA
 Then VLCF acyl-CoA is transported into
peroxisome by ABCD (ATP-binding cassette
transporters of subfamily D)
 There are 4 reaction of 𝛽-oxidation in
peroxisomes like 𝛽-oxidation in mitochondria
except for the first reaction
o In mitochondria : Acyl-CoA convert by fatty acyl
CoA dehydrogenase to :
• Trans-2-Enoyl CoA
• FADH2  ETC
o In peroxisome : Acyl-CoA is catalyzed by a FAD-
containing acyl CoA oxidase to :
• Trans-2-Enoyl CoA
• FADH2  is oxidized by O2, which is
reduced to hydrogen peroxide (H2O2)
• H2O2 is reduced to H2O by catalase
 The products of 𝛽-oxidation in peroxisomes transport
to mitochondria for further metabolism
 Acetyl-CoA
 NADH
 Propionyl-CoA
 Shorten acyl-CoA
By: Savath Sovannak
  Alpha oxidation is a process in which FA is
shortened by one carbon atom from 𝛼 position in
peroxisome
 It is a mechanism mainly for oxidation of branched
chain fatty acid, which are methylated at beta
position.
 It is specific for oxidation of phytanic acid (3, 7, 11,
15 tetra methyl palmitic acid)
 The reaction does not require CoASH and does not
generate ATP :
 First Phytanic Acid is activated like VLCFA and
transported into peroxisome by ABCD like
VLCFA
 Phytanoyl-CoA convert to 2 (or 𝛼)-
hydroxyphytanoyl-CoA by phytanoyl CoA
alpha-hydroxylase (PhyH)
 Then 2-hydroxyphytanoyl-CoA covert to
pristanal by 2-hydroxyphytanoyl-CoA lyase
 Pristanal convert to Pristanoic acid by
aldehyde dehydrogenase
 Pristanoic acid is activated (+ CoA) and undergoes beta-oxidation  6 cycle of beta-oxidation
 form 3 Acetyl CoA, 3 Propionyl CoA, and 2-methyl-propionyl-CoA

By: Savath Sovannak

 It is oxidation of terminal CH3 group (𝜔 carbon) of fatty
acid by enzymes of smooth endoplasmic reticulum 
produces dicarboxylic fatty acids
 It is minor pathway for FA oxidation
 There are 3 steps of omega oxidation :
 First of all, FA is hydroxylated by P450 mixed-
function oxidase  form fatty alcohol
 Then fatty alcohol is oxidized to fatty aldehyde by
alcohol dehydrogenase
 Fatty aldehyde is oxidized to fatty dicarboxylic acid
by aldehyde dehydrogenase
 Then fatty dicarboxylic acid can undergo beta oxidation
in either peroxisome or mitochondria depend the length
of FA

By: Savath Sovannak