Synthesis of Long-chain Fatty Acids

Formerly considered

Reversal of oxidation within the mitochondria

Fatty acids are synthesized by an

Now

extramitochondrial system Responsible for complete synthesis of Palmitate from Acetyl-CoA in the Cystosol

Glucose

Most mammals Primary substrate for lipogenesis Ruminants (Mammals with four-chambered stomachs and two-toed foot) Main fuel molecule produced by the diet Primary substrate

Acetate

Present in: Liver, kidney, adipose tissue, and

lactating mammary glands.

Site: Cytosol/ Cytoplasm of the cell

Carried out by two enzyme systems:

Acetyl-CoA Carboxylase Fatty Acid Synthase

1.

Production of Acetyl-CoA and NADPH Biosynthesis of Malonyl-CoA Reactions of Fatty Acid Synthase Complex

2.

3.

Sources of NADPH:
Pentose Phosphate Pathway
Main source Its oxidative reactions are the chief source of the hydrogen required for the reductive synthesis of fatty acids.

Malate enzyme
NADP Enzyme dehydrogenase Enzyme in the conversion of malate to pyruvate

Isocitrate dehydrogenase
Probably not a substantial source.

Acetyl-CoA is the principal building block of Fatty Acids.

Formed from carbohydrate (eg. Glucose) via the oxidation of pyruvate within the mitochondria. Citric acid cycle

Source of the 2 carbon sub-units added to the growing FA chain on FA synthase complex Requirements:

Bicarbonate (for the initial reaction for the carboxylation of Acetyl-CoA to Malonyl-CoA) ATP Enzyme: Acetyl-CoA Carboxylase
3 Components of Acetyl-CoA carboxylase:

1.Biotin carboxylase 2.Biotin carboxylase carrier protein 3.Transcarboxylase Co-enzyme: Biotin (the enzyme prosthetic group)

Transfer of carboxyl to Acetyl-CoA to Malonyl-CoA

Catalyzed by: Acetyl-CoA carboxylase

O CH3- C S-CoA
-OOC-CH 2

C S-CoA

Acetyl-CoA
Enz-BiotinEnz-Biotin- *COO *COO-

Malonyl-CoA
Enz- Biotin

ADP + Pi

* -ATP ATP + + H*CO HCO 3 3

Carboxylation of Biotin involving ATP

Fatty Acid Synthase Complex

A polypeptide containing seven enzyme activities. A dimer of two identical polypeptide monomers, 1 and 2, each consisting of: Seven enzyme activities:
a) b) c) d) e) f) g) Ketoacyl synthase Acetyl transacylase Malonyl transacylase Hydratase Enoyl reductase Ketoacyl reductase Thioesterase

Acyl carrier protein (ACP)

Enzyme: Acetyl transacylase

O CH3 C

~S CoA

HSPan

1 CysHS
ACETYL TRANSACYLASE

HSCys

2 PanHS

HS CoA

CysS

O CCH3

2 PanS

Enzyme: Malonyl transacylase

HSPan

1 CysHS

HOOCCH2C
HSCys

S CoA

2 PanHS

Malonyl Transacylase O CCH3 CCH2COOH O

HS CoA

CysS ~

2 PanS ~

Acetyl (acyl)malonyl enzyme

O
1

Enzyme: 3-ketoacyl synthase

Acetyl group
Methylene group

CysS

CCH3 CCH2COOH

2 PanS

O
3-ketoacyl synthase CO2
1

CysSH

3-Ketoacyl enzyme (acetoacetyl enzyme) O

2 PanS

CCH2 CCH3 O

Enzyme: 3- ketoacyl reductase

1

CysSH

O 2 PanS CCH2CCH3 O

NADPH
NADP+
1

3-Ketoacyl enzyme (acetoacetyl enzyme)

3-Ketoacyl Reductase

CysSH

OH
2 PanS CCH2 CHCH3 O -hydroxyacyl enzyme

CysSH

Enzyme: Hydratase
OH CCH2COOH CHCH3 -hydroxyacyl enzyme

2 PanS

O
H2O
1

Hydratase

CysSH

2 PanS

CCH CHCH3 O 2,3- Unsaturated acyl enzyme

CysSH

Enzyme: Enoyl reductase

2 PanS

CCH CHCH3
O

2,3- Unsaturated acyl enzyme

NADPH
Enoyl Reductase

NADP+
1

CysSH

Saturated acyl enzyme CCH2CH2CH3 O

2 PanS

The sequence of reactions:

Repeated 6 more times New malonyl residue being incorporated during each sequence until a saturated 16-carbon acyl radical (palmityl) has been assembled. Enzyme: Thioesterase (deacylase)

CH2CO S CoA + 7HOOCCH2CO S CoA + 14NADPH + 14H+ CH3(CH2)14COOH +7CO2 + 6H2O + 8CoA SH + 14NADP+

Two centers of activity in one dimer complex that function independently and simultaneously.

http://oregonstate.edu/dept/biochem/hhmi/ hhmiclasses/biochem/lectnoteskga/2kjan19l ecturenotes.html Harpers Biochemistry, 25th Edition