5.1.3.

1 BIOSYNTHESIS OF FATTY ACIDS : LIPOGENESIS

THERE ARE SOME PATHWAYS OF FATTY ACIDS BIOSYNTHESIS : 1. Biosynthesis of fatty acids from acetyl CoA ( de novo ). Its occured outer ( extra ) mitochondria 2. Elongation of fatty acid chains Biosynthesis of fatty acids from lower fatty acids. For example : Biosynthesis palmitic acids ( C16 ) from decanoic acids ( C10 ) Its occured in mitochondria and microsome

FATTY ACID BIOSYNTHESIS ( DE NOVO )

The first steps of biosynthesis is forming malonyl CoA from acetyl CoA, catalyzed by acetyl CoA carboxylase enzyme ( regulatory enzyme ). This enzyme is induced by insulin, citrate and isocitrate. So in Diabetes Mellitus, biosynthesis of fatty acid is very decrease. Then followed by forming acetyl enzyme from another acetyl CoA. Acetyl enzym ( C2 ) then condensed with malonyl CoA to form acyl malonyl enzyme. Acyl malonyl enzyme is carboxylated by condensing enzyme to form keto acyl-enzyme and then reduced by keto acyl enzyme reductase to form D (+) hydroxyl acyl enzyme. D (+) hydroxy acyl enzyme is hydrolated by hydratase enzyme to form trans -

unsaturated acyl enzyme and then reduced ones more by , -unsaturated acyl enzyme reductase to form acyl enzym ( C4 ). Acyl enzym ( C4 ) then condensed with malonyl CoA and reaction return again, like reaction above to form acyl enzym ( C6 ) and so on C8 C10 C16 ( Palmityl enzym ). Palmityl enzym is hydrolated to form palmitic acid by palmityl-S-enzym deacylase.

ACETYL CoA FROM MITOCHONDRIAL MATRIX ENTERS CYTOSOL IN ORDER TO HIGHER FATTY ACID BIOSYNTHESIS

Acetate Transport from Mitochondria to Sitoplasm

Matrix
Tricarboxilate Transporter Glucose CoA-SH Pyruvate Acetyl-CoA Amino acids Citrate Citrate ATP

Inner membrane

Outer membrane

Cytosol
CoA-SH Fatty acid synthesis ADP+P1 Acetyl-CoA

cs

cl

Oxaloacetat NADH + H+ NAD+

Oxaloacetate NADH + H+

md
Malate

md
NAD+ Malate NADP+

ADP+P1 ATP CO2

pc

me
Malate Pyruvate -ketoglutarate transporter NADPH + H+ Pyruvate CO2 cs cl md Me pc = citrate synthase = citrate lyase = malat dehydrogenase = malat enzime = pyruvate carboxylase

Pyruvate transporter

Acetyl CoA binds oxalo acetate to form citrate. Citrate out from mitochondrial matrix enters cytosol. In the cytosol citrate splits to oxalo acetate and acetyl CoA and ready for synthesis higher fatty acid.

POST PALMITATE SYNTHESIS

Palmitate is activated by ATP and CoA, catalyzed by acyl CoA synthetase enzyme to form palmityl-CoA. Palmityl CoA may be esterified with glycerol or cholesterol to form acyl glycerols or cholesteryl esters, or may be elongated or desaturated to form higher fatty acid like stearate or oleate, or unsaturated fatty acids like palmitoleate.

BIOSYNTHESIS OF FATTY ACID DERIVATE

Conversion of Arachidonate to Prostaglandin

Vit. E

ASPIRIN INDOMETHACINE

__

Polyunsaturated fatty acids like linoleate, linolenate or arachidonate can synthesis some fatty acid derivates like prostaglandins, thromboxans, leucotriens and lipoxins. They are pharmacologic active substances in human bodies.

PHYSIOLOGIC EFFECT OF PROSTAGLANDINS

- Uterus contraction * Family planning * Induction of labor * Termination of labor - Decrease cAMP in adipose tissues So induce lipogenesis - Thromboxan : * Vaso constraction of blood vessels Thrombocyt aggregation Intra vascular agglutination Thrombus - Prostasiklin : * Suppress thrombocyt aggregation so blood agglutination not occured - Leucotrien * Bronchus contraction * Slow reacting anaphylaxis ( SRS A )

5.1.3.2. OXIDATION OF FATTY ACIDS & KETOGENESIS Fatty acids oxidation ( oxidation )

Fatty Acids Transport from Sitoplasm to Mitochondria

O R C S-CoA

Intermembrane space O Carnitine O R C Carnitine R C Carnitine Carnitine O CoA-S R C S-CoA

CoA-SH

Carnitine acyltranferase I

Matrix

Fatty acids are activated by ATP and CoA-SH, catalyzed by thiokinase enzym to form acyl-CoA (active fatty acids). This reaction, occured outer mitochondria. Acyl-CoA binds carnithin (lipids transporter) to form acyl carnithin, and then enter mitochondrial matrix. Acyl-carnithin splits into carnithin and acyl-CoA in mitochondria and ready to be oxidized (-oxidation).

FATTY ACIDS OXIDATION ( OXIDATION )

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Fatty acids oxidation occur in mitochondrial matrix. Acyl CoA (Cn) is reduced to form (trans) , -unsaturated acyl CoA catalyzed by acyl CoA dehidrogenase enzyme and flavoprotein (Fp) as coenzyme, then hydrolyzed by enoyl hidrase enzyme to form L(+) -hydroxy acyl CoA. Then reduced ones more by L(+) hydroxy acyl CoA dehydrogenase enzyme and NAD+ as coenzyme to form keto acyl CoA. keto acyl CoA is splited by thiolase enzyme to form acyl CoA (Cn-2) + acetyl CoA and enters TCA cycle. Acyl CoA (Cn-2) return to be oxidized like reaction above and one mol acetyl CoA is resulted + acyl CoA (Cn-4) and so on, until all of C atoms of acyl CoA splited into several mols of acetyl CoA. Acetyl CoA are formed = n/2 mols. oxidation times = n/2 1 times n = numbers of C atoms of fatty acids. All of acetyl CoA enter the TCA cycle to form very much energy ( ATP ).

THREE STAGES OF FATTY ACID OXIDATION

1. Forming acetyl CoA ( oxidation ) 2. Acetyl CoA enters TCA cycle forming FADH2 and NADH + H+ 3. FADH2 and NADH enter respiratory chains forming energy (ATP)

Energy are generated from oxidation of 1 mol palmitate : - 7 x oxidation - 8 mol acetyl CoA (TCA) 35 ATP 96 ATP 131 ATP Energy are needed (1 ATP activation + 1 ATP enters mitochondria) 1 mol FADH2 ~ 2 ATP 1 mol NADH ~ 3 ATP 2 ATP 129 ATP

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REGULATION OF FATTY ACIDS OXIDATION IN THE LIVER

During oxidation of fatty acids, acyl CoA inhibits acetyl CoA carboxylase enzyme (regulatory enzyme for fatty acid biosynthesis). So fatty acid biosynthesis can not occurred during oxidation. On the other hands, during fatty acid biosynthesis, malonyl CoA inhibits carnithin palmitoyl transferase enzyme, so fatty acid (acyl CoA) can not enter mitochondria, and oxidation of fatty acid can not occured during biosynthesis of fatty acids.

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KETON BODIES METABOLISM Formation of keton bodies in the liver

In order to fatty acids oxidation ( oxidation), aceto acetyl CoA is condensed with acetyl CoA, forming hydroxy -methyl glutaryl CoA (HMG-CoA). HMG CoA splits to acetyl CoA and aceto acetate, catalyzed by HMG-CoA lyase enzyme. Aceto acetate is reduced to form hydroxy butyrate by -hydroxy butyrate dehydrogenase enzyme. Aceto acetate also decarboxylated forming acetone by aceto acetate decarboxylase enzyme. Aceto acetate, hydroxy butyrate and aceton are called keton bodies.

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THREE KINDS OF KETON BODIES

Keton bodies : 1. Aceto acetate 2. hydroxy butyrate 3. Acetone 1 and 2 are acids

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EXCRETION & CATABOLISM OF KETON BODIES

Keton bodies are formed in the liver, and to be excreted in the lung (acetone) and urine (aceto acetate & acetone). Keton bodies catabolism occured in extra hepatic tissues to form acetyl CoA and enters TCA cycle. So keton bodies are energy resources too. In well feed condition, keton bodies blood level is not more than 1mg% and in the urine is not more than 1 gr per 24 hours. In Diabetes Mellitus, keton bodies catabolism in extra hepatic tissues decrease, and acetyl CoA almost can not enter TCA cycle, so keton bodies blood level very increase, called ketosis or keto acidosis.

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CATABOLISM OF KETON BODIES IN EXTRA HEPATIC TISSUES 1. Aceto acetate CoA transferase Aceto acetate Succinyl CoA Aceto acetyl CoA Succinate Thiolase Aceto Acetyl CoA Acetyl CoA

TCA 2. hydroxy butyrate D(-)-OH-butyrate dehydrogenase D(-)-hydroxy Aceto acetate butyrate NAD+ NADH + H+ Aceto acetyl CoA Acetyl CoA 3. Acetone : Very difficult to oxidize TCA

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KETOSIS (KETO ACIDOSIS) Can be found in several conditions : 1. Diabetes Mellitus Energy supply failure from carbohydrate (glucose) metabolism FFA -oxidation lypolysis

FFA -oxidation CHOLESTEROL ACETYL KoA HMG-CoA reductase HMG-CoA HMG-CoA lyase TCA KETON BODIES Acetyl CoA (extra hepatic tissues) KETO ACIDOSIS TCA

2. Starvation * Glucose are oxidized only for erythrocytes and Central Nervous System (CNS) which are depending on glucoses supply * Energy resources will be taken from lipids storage lipolysis FFA

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FFA -oxidation ACETYL CoA CHOLESTEROL HMG-CoA TCA KETON BODIES

ACETYL CoA (extra hepatic tissues)

SLIGHT KETOSIS

TCA

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3. High fat and low carbohydrate diet Similar to fasting condition

KETOGENESIS REGULATION IN THE LIVER

Ketogenesis pathway, depend on carnithin palmitoyl transferase enzyme activity (CPT). So in well feed condition, ketogenesis is decrease and in fasting conditions and Diabetes Mellitus ketogenesis is increase, because CPT enzyme activity increase.

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5.1.3.3. METABOLISM OF TRIACYL GLYCEROLS & PHOSPHOLIPIDS

Receptors catch hormones (epinephrin). Adenylate cyclase is activated. ATP is changed to cAMP. cAMP activates protein kinase enzyme and induce lipolysis, so Triacyl Glycerols split into glycerols and fatty acids. Fatty acids enter blood stream and to be transported to several tissues for oxidized. Glycerol is taken by the liver to form glucose (gluconeogenesis), or forming glyceroldehyde 3-phosphates, then enter glycolysis pathway.

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TRIACYL GLYCEROLS BIOSYNTHESIS

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Glycerol-3-phosphate is esterified with acyl CoA (active fatty acids) to form lysophosphatidic acid, catalyzed by acyl transferase enzyme. Lysophosphatidic acids is esterified again with acyl CoA to form phosphatidic acid, catalyzed by the same enzyme. Then phosphatidic acid is changed to diacyl glycerol, catalyzed by phosphatidate phosphatase enzyme. And the last reaction, diacyl glycerol is esterified again with acyl CoA, to form Triacyl glycerol. BIOSYNTHESIS OF CERAMIDE

Ceramide is synthesized from palmitate and amino acid serin.

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BIOSYNTHESIS OF SPHINGOMEYLIN

BIOSYNTHESIS OF SULFATIDE

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METABOLISM OF ADIPOSE TISSUE

Triacyl glycerol is splited to free fatty acids (FFA) and glycerol by hormon sensitive lipase enzyme. A part of FFA is activated to acyl CoA in adipose tissue, and the other part enter blood stream. Acyl CoA is oxidized to acetyl CoA and enter citric acid cycle (TCA) to form energy (ATP), CO2 and H2O. Glycerol in adipose tissue can not be activated to glycerol 3 phosphate, because there is not glycerokinase enzyme activity in adipose tissue. So glycerol enters blood stream to be taken by the liver to form glucose (gluconeogenesis) or enter glycolysis pathway in the liver, or esterified with FFA forming triacyl glycerol. In order to biosynthesis of triacyl glycerol in adipose tissue, glucose from the blood, enters adipose tissue and in glycolysis pathway, dihydroxy acetone phosphate is formed and to be changed to glycerol 3 phosphate (active glycerol). Triacyl glycerol is synthesized from acyl CoA and glycerol 3 phosphate.

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REGULATION OF LYPOLYSIS IN ADIPOSE TISSUE

Some hormones like epinephrine, nor epinephrine, ACTH, TSH, and glucagon activate adenylyl cyclase enzyme to change ATP to cAMP. cAMP induce cAMP dependent protein kinase enzyme to activate hormon sensitive lipase enzyme. So triacyl glycerols split into glycerols and free fathy acids. On the other hand, insulin suppress adenylyl cyclase, enzyme or induce phospho diesterase enzyme, so cAMP decrease and lipolysis is suppressed or lipogenesis is induced.

5.1.3.4. LIPID TRANSPORT & STORAGE Lipids are transported in lipoprotein form. There are 4 fracsions of lipoprotein : 1. 2. Chylomicron (derived from intestinal absorpsion of triacyl glycerol) Verry low density lipoprotein ( VLDL ), derived from the liver for the export of triacyl glycerol

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3. 4.

Low density lipoprotein (LDL), representing a final stage in the catabolism of VLDL High density lipoprotein (HDL), involved in chylomicron and VLDL metabolism and cholesterol transport.

METABOLISM OF CHYLOMICRON

Chylomicron derived from intestinal absorpsion of triacyl glycerol. TAG content of chylomicron is splited into glycerol and free fatty acid (FFA) catalyzed by lipoprotein lipase enzyme. FFA is taken by extra hepatic tissues to oxidize or reesterification. Glcyerol is taken by the liver. Lipoprotein lipase enzyme is induced by insuline hormon. So in Diabetes Mellitus lipoprotein lipase activity decrease and triacyl glycerols accumulate in the blood (hyper triglyceridemia).

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METABOLISM OF VLDL

VLDL derived from the liver. As chylomicron triacyl glycerol content of VLDL is splited into glycerol and free fatty acid (FFA) by lipoprotein lipase enzyme. FFA is taken by extra hepatic tissues to oxidize or reesterication . Glycerol is taken by the liver. Lipoprotein lipase enzyme is induced by insulin, so in Diabetes Mellitus, hypertriglyceridemia is occured. LDL is final stage in the catabolism of VLDL. And LDL split in the liver and extra hepatic tissues.

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LIPID STORAGE DISEASE

Fatty livers fall into two main categories : 1. Associated with raised levels of plasma free fatty acids, resulting from mobilization of fat from adipose tissue or from the hydrolysis lipoprotein lipase in extra hepatic tissue. Increasing amounts of free fatty acids are taken by the liver and esterified. The production of VLDL does not keep pace with the influx of free fatty acids, allowing triacyl glycerol to accumulate, causing a fatty liver. This condition can occured in uncontrolled Diabetes Mellitus, starvation and the feeding of high fat diet low carbohydrate. 2. Due to a metabolic block in the production of plasma lipoproteins, thus allowing triacyl glycerol to accumulate. This condition may be occured in : a). A block in apoprotein synthesis b). A block in the synthesis of the lipoprotein from lipid and apo lipoprotein c). A failure in provision of phospholipids that are found in lipoprotein d). A failure in secretary mechanism itself

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5.1.3.5. METABOLISM OF CHOLESTEROL : BIOSYNTHESIS, TRANSPORT & EXCRETION

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Synthesis of Squalene
CH3 OOC CH2 C CH2 MEVALONATE PROSES * CH 2 * CH 2 Squalene
There are 5 stages of cholesterol biosynthesis : 1. Biosynthesis of mevalonate from acetyl CoA This stage is very important, because there is regulatory enzyme HMGCoA reductase. This enzyme is suppressed by bile acid, cholesterol, mevalonate, statins eg mevastatin and lovastatin. Mevastatin and lovastatin are used as HMG CoA reductase inhibitor to inhibit cholesterol biosynthesis in hypercholesterolemia states : 2. 3. 4. 5. Biosynthesis of active isoprenoid from mevalonate Biosynthesis squalene from six units of isoprenoid Biosynthesis of lanosterol from squalene Biosynthesis cholesterol from lanosterol

OH CH2 OH

Cholesterol Transport Cholesterol is transported in lipoprotein form especially in low density lipoprotein (LDL). A little part of cholesterol also transported in chylomicron, VLDL and HDL. Cholesterol excretion : Cholesterol is excreted in the bile as :

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1. 2. 3.

Cholesterols Bile acids Bile salts

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REFERENCE 1. Harper, H.A : Biochemistry. Fifth edition, 2000, 230 285. A Lange Medical Book. 2. Lehninger A.L.: Principles of Biochemistry. Third edition, 2000, 363 386. Worth Publisher

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