3 fa synthesis
TRANSCRIPT
![Page 1: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/1.jpg)
Fatty acid synthesis
![Page 2: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/2.jpg)
Glycerol-P Glycerol
Triacylglycerol
Fatty acyl CoA Fatty acid
Malonyl CoA
Acetyl CoA
Glucose
Pyruvate
TCA cycle
Fed state
![Page 3: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/3.jpg)
Glycerol-P Glycerol
Triacylglycerol
Fatty acyl CoA Fatty acid
Malonyl CoA
Acetyl CoA
Glucose
Pyruvate
TCA cycle
Starved state
gluconeogenesis
![Page 4: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/4.jpg)
Fatty acid biosynthesis
![Page 5: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/5.jpg)
A three carbon intermediate, malonyl-CoA, initiates fatty acid synthesis
• FA biosynthesis and breakdown occur by different pathways and take place in different parts of the cell.
• Biosynthesis requires malonyl-CoA
Glycerol-P
Triacylglycerol
Fatty acyl CoA
Malonyl CoA
Acetyl CoA
Glucose
Pyruvate
TCA cycle
![Page 6: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/6.jpg)
The carboxylation of acetyl-CoA yields malonyl-CoA
CH3-C-S-CoA
=O
HCO3-
-OOC-CH2-C-S-CoA
=O
Acetyl-CoA
Malonyl-CoA
Acetyl CoA carboxylase
![Page 7: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/7.jpg)
Assembly of a long chain fatty acid
• Once malonyl-CoA is synthesized, long carbon FA chains may be assembled in a repeating four-step sequence.
• With each passage through the cycle the fatty acyl chain is extended by two carbons.
• When the chain reaches 16 carbons, the product palmitate (16:0) leaves the cycle.
![Page 8: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/8.jpg)
The first round of FA biosynthesis
• To initiate FA biosynthesis, malonyl and acetyl groups are activated on to the enzyme fatty acid synthase.
H
H
Malony-CoA
Acetyl-CoA
+
![Page 9: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/9.jpg)
Step 1.
• Condensation of an activated acyl group and two carbons derived from malonyl-CoA
![Page 10: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/10.jpg)
Step 2.
• The -keto group is reduced to an alcohol by NADPH
![Page 11: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/11.jpg)
Step 3.
• The elimination of water creates a double bond.
![Page 12: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/12.jpg)
• The double bond is reduced to form the corresponding saturated fatty acyl group.
Step 4.
![Page 13: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/13.jpg)
Repetition of these four steps leads to fatty acid synthesis
• When reaches 16 carbons, the product leaves the cycle.
• All the reactions in the synthetic process are catalyzed by a multi-enzyme complex, fatty acid synthase.
![Page 14: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/14.jpg)
A more detailed look at fatty acid synthase
![Page 15: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/15.jpg)
Fatty acyl synthase contains six enzymatic activities
• Each segment of the disk represents one of the six enzymatic activities of the complex.
• At the center is the ACP – acyl carrier protein - with its phosphopantetheine arm ending in –SH.
![Page 16: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/16.jpg)
The function of the prosthetic group of the ACP
• Serve as a flexible arm, tethering the growing fatty acyl chain to the surface of the synthase complex
• Carrying the reaction intermediates from one enzyme active site to the next.
![Page 17: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/17.jpg)
Activation of acetyl and malonyl groups
• Before Steps 1-4, the two thiol groups on the enzyme complex must be charged with the correct acyl groups.
![Page 18: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/18.jpg)
• The acetyl group from acetyl-CoA is transferred to the Cys-SH group of the -ketoacyl ACP synthase.
• This reaction is catalyzed by acetyl-CoA transacetylase.
The activation of the acetyl group
![Page 19: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/19.jpg)
The activation of the malonyl group
• Transfer of the malonyl group to the –SH group of the ACP is catalyzed by malonyl-CoA ACP transferase.
• The charged acetyl and malonyl groups are now in close proximity to each other
![Page 20: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/20.jpg)
• Condensation of the activated acetyl and malonyl groups to form acetoacetyl-ACP, catalyzed by -ketoacyl-ACP synthase.
Step 1.
![Page 21: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/21.jpg)
Step 2.• Reduction. The acetoacetyl-ACP is reduced to -
hydroxybutyryl-ACP, catalyzed by ketoacyl-ACP reductase (needs NADPH + H+)
![Page 22: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/22.jpg)
Step 3.• Dehydration to yield a double bond in the product, trans-
2-butenoyl-ACP, catalyzed by hydroxyacyl-ACP dehydratase.
![Page 23: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/23.jpg)
Step 4.
• Reduction of the double bond to form butyryl-ACP, catalyzed by enoyl-reductase.
• Another NADPH dependent reaction.
![Page 24: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/24.jpg)
The growing chain is transferred from the acyl carrier protein
• This reaction makes way for the next incoming malonyl group.
• The enzyme involved is acetyl-CoA transacetylase.
![Page 25: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/25.jpg)
Beginning of the second round of the FA synthesis cycle
• The butyryl group is on the Cys-SH group.
• The incoming malonyl group is first attached to ACP.
• In the condensation step, the entire butyryl group is exchanged for the carboxyl group on the malonyl residue.
![Page 26: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/26.jpg)
The result of fatty acyl synthase activity
• Seven cycles of condensation and reduction produce the 16-carbon saturated palmitoyl group, still bound to ACP.
• Chain elongation usually stops at this point, and free palmitate is released from the ACP molecule by hydrolytic activity in the synthase complex.
• Smaller amounts of longer fatty acids such as stearate (18:0) are also formed.
![Page 27: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/27.jpg)
The overall reaction for the synthesis of palmitate from acetyl-CoA can be considered in two parts.
![Page 28: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/28.jpg)
Part 1.
• First, the formation of seven malonyl-CoA molecules:
7Acetyl-CoA + 7CO2 + 7ATP 7malonyl-CoA + 7ADP + 7Pi
![Page 29: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/29.jpg)
Part 2.
• Then the seven cycles of condensation and reduction
Acetyl-CoA + 7malonyl-CoA + 14NADPH + 14H+
palmitate + 7CO2 + 8CoA + 14NADP+ + 6H2O
• The biosynthesis of FAs requires acetyl-CoA and the input of energy in the form of ATP and reducing power of NADPH.
![Page 30: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/30.jpg)
Location of FA synthesis
• FA synthase complex is found exclusively in the cytosol.
• The location segregates synthetic processes from degradative reactions.
![Page 31: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/31.jpg)
In hepatocytes:
the [NADPH]/[NAD+] ratio is very high (~75) in the cytosol, furnishing a strongly reducing environment for the reductive synthesis of fatty acids and other biomolecules.
![Page 32: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/32.jpg)
Fatty acid synthesis requires considerable amounts of NADPH + H+
Acetyl-CoA + 7malonyl-CoA + 14NADPH + 14H+
palmitate + 7CO2 + 8CoA + 14NADP+ + 6H2O
• In hepatocytes and adipocytes, cytosolic NADPH is largely generated by the malic enzyme and by the pentose phosphate pathway.
![Page 33: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/33.jpg)
1. The malic enzyme
• The pyruvate produced in the reaction reenters the mitochondrion.
![Page 34: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/34.jpg)
2. The pentose phosphate pathway
• In hepatocytes and the mammary gland of lactating animals, the NADPH is supplied primarily by the pentose phosphate pathway.
![Page 35: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/35.jpg)
Fatty acid synthesis requires considerable amounts of acetyl-CoA
7Acetyl-CoA + 7CO2 + 7ATP 7malonyl-CoA + 7ADP + 7Pi
• Nearly all acetyl-CoA used in fatty acid synthesis is formed in mitochondria from pyruvate oxidation.
• So acetate must go from the mitochondria to the cytosol
Mitochondria – site of acetate manufacture
Cytosol – site of acetate utilization
![Page 36: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/36.jpg)
Acetate is shuttled out of mitochondria as citrate
• The mitochondrial inner membrane is impermeable to acetyl-CoA
• Intra-mitochondrial acetyl-CoA first reacts with oxaloacetate to form citrate, in the TCA cycle catalyzed by citrate synthase.
![Page 37: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/37.jpg)
• Citrate then passes into the cytosol through the mitochondrial inner membrane on the citrate transporter.
• In the cytosol, citrate is cleaved by citrate lyase regenerating acetyl-CoA.
![Page 38: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/38.jpg)
• The other product --oxaloacetate cannot return to the mitochondrial matrix directly.
• Instead, oxaloacetate is reduced to malate
![Page 39: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/39.jpg)
• Malate returns to the mitochondrial matrix on the malate--ketoglutarate transporter in exchange for citrate.
![Page 40: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/40.jpg)
Regulation of fatty acid synthesis
• When a cell has more energy, the excess is generally converted to FAs and stored as lipids such as triacylglycerol.
• The reaction catalyzed by acetyl-CoA carboxylase is the rate limiting step in the biosynthesis of fatty acids.
![Page 41: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/41.jpg)
The carboxylation of acetyl-CoA yields malonyl-CoA
CH3-C-S-CoA
=O
HCO3-
-OOC-CH2-C-S-CoA=O
Acetyl-CoA
Malonyl-CoA
![Page 42: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/42.jpg)
Regulation of acetyl-CoA carboxylase(1)
• Palmitoyl-CoA acts as a feedback inhibitor of the enzyme, and citrate is an activator.
• When there is an increase in mitochondrial acetyl-CoA and ATP, citrate is transported out of mitochondria,
• Citrate becomes both the precursor of cytosolic acetyl-CoA and a signal for the activation of acetyl-CoA carboxylase.
![Page 43: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/43.jpg)
Regulation of acetyl-CoA carboxylase (2)
![Page 44: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/44.jpg)
Regulation of acetyl-CoA carboxylase (3)
• Additionally, these pathways are regulated at the level of gene expression.
• For example, when animals ingest an excess of certain polyunsaturated fatty acids, the expression of genes encoding a wide range of lipogenic enzymes in the liver is suppressed.
![Page 45: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/45.jpg)
Additional modification to the newly synthesized fatty acid
• Extended to form longer fatty acids
• Converted to monounsaturated and polyunsaturated fatty acids
![Page 46: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/46.jpg)
Fatty acid elongation
• Palmitate in animal cells is the precursor of other long-chained FAs.
• By further additions of acetyl groups, through the action of FA elongation systems present in the smooth endoplasmic reticulum and the mitochondria.
![Page 47: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/47.jpg)
The desaturation of FAs
• Palmitate and stearate serve as precursors of the two most common monosaturated fatty acids of animal cells: palmitoleate (16:19), and oleate (18:19).
• The double bond is introduced by fatty acyl-CoA desaturase in the smooth endoplasmic reticulum.
![Page 48: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/48.jpg)
• Mammalian hepatocytes readily introduce double bonds at the 9 position of FAs but cannot between C-10 and the methyl-terminal end.
• Linoleate, 18:29,12 and linolenate 18:39,12,15 cannot be synthesized by mammals, but plants can synthesize both.
Essential fatty acids
![Page 49: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/49.jpg)
The fate of fatty acids
• Most of the FAs synthesized or ingested by an organism have one of two fates:
i. incorporated into triacylglycerols for the storage of metabolic energy
ii. incorporation into the phospholipid components of membranes.
![Page 50: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/50.jpg)
The formation of phosphatidic acid
• Fatty acyl groups are first activated by formation of fatty acyl-CoA molecules.
• then transferred to ester linkage with L-glycerol 3-phosphate.
![Page 51: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/51.jpg)
Phosphatidic acid may be converted to triacylglycerols or phospholipids
• Triacylglycerols and phosholipids are both synthesized from phosphatidic acid
![Page 52: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/52.jpg)
Lecithin (phosphatidyl choline)
O
O
H2C O C RO
R C O CH
H2C O P O CH2 CH2 N+ CH3
CH3
CH3
O_
Phosphatidic Acid Choline
![Page 53: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/53.jpg)
Partitioning of the fates of fatty acids
• Depends on the needs of the organism:
• During rapid growth, synthesis of new membranes requires membrane phospholipid synthesis
• Organisms that have a plentiful supply of food but are not actively growing shunt most of their fatty acids into storage fats.
![Page 54: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/54.jpg)
Summary of lipid metabolism
• FA biosynthesis requires malonyl-CoA formation• The long carbon chains of FA acids are assembled in
a repeating four-step sequence catalyzed by the multifunctional enzyme fatty acid synthase.
• With each passage through the cycle, the fatty acyl chain is extended by two carbons
• When the chain length reaches 16 carbons, the product (palmitate 16:0) leaves the cycle.
![Page 55: 3 FA Synthesis](https://reader035.vdocuments.mx/reader035/viewer/2022081514/5537b4504a79591c7b8b45c7/html5/thumbnails/55.jpg)
• Cytosolic NADPH is largely generated by the malic enzyme and by the pentose phosphate pathway.
• FA biosynthesis occurs in the cytosol• FA biosynthesis is regulated by the activity
of acetyl-CoA carboxylase• Synthesized FA are either stored as TG or
made into membrane lipids