Lipid Biochemistry

Aulanniam Biochemistry Laboratory Chemistry Department Brawijaya University

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Lipids: Hydrophobic molecules

Fats (animal) and Oils (plant) - energy storage, insulation

Fatty acid - Long hydrocarbon tail with carboxly -COOH group at the head. Saturated - no double bonds; saturated with H Unsaturated - one or more H replaced by double bond - stays liquid Carboxyl groups on fatty acid link to -OH group on a 3-carbon alcohol (glycerol)

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A fatty acid

Aulani "Biokimia" Presentation 6

Building a fat molecule

Aulani "Biokimia" Presentation 6

 Source of stored energy in living organisms Lipids contain the elements carbon, hydrogen,

and oxygen Glycerol and fatty acids are the building blocks of lipids Examples of lipids are fats and oils

fatty acid

glycerol

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For simplicity, the fatty acids will be abbreviated as:

HOOC-R
where "R" simply represents the long carbon chain. glycerol + 3 fatty acids = a fat or oil

makes a fatty acid and 3 water molecules, 3 H2O Since a fat or oil contains 3 fatty acid units, they are sometimes called triglycerides
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Saturated and unsaturated fats

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Phospholipids
One fatty acid replaced by phosphate PO4 Molecule has Hydrophilic head, and long

hydrophobic tail. Fatty acids unsaturated- remains fluid Main component of cell membranes

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Aulani "Biokimia" Presentation 6

Most Common Fatty Acids in Di- and Triglycerides

Fatty acid Myristic Carbon:Double bonds 14:0 Double bonds

Palmitic
Palmitoleic Stearic Oleic

16:0
16:1 18:0 18:1 Cis-9 Cis-9

Linoleic
Linolenic Arachidonic Eicosapentaenoic

18:2
18:3 20:4 20:5

Cis-9,12
Cis-9,12,15 Cis-5,8,11,14 Cis-5,8,11,14,17

Docosahexaenoic

22:6

Cis-4,7,10,13,16,19

CH3(CH2)nCOOH
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Linolenic Acid
Omega-3

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Cell membrane- Phospholipid bilayer

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Predominant Energy Pathways

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We use fat in the form of triglyceride (3 fatty acids and 1 glycerol).

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Fat Metabolism
Mostly handled by the liver
Fats must first be broken down to form

acetic acid which is subsequently oxidized. Oxidation (breakdown) of fats is not always complete. Intermediate products accumulate in the blood causing the blood to become acidic (acidosis or ketosis)

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Cholesterol
Structural basis of steroid hormones and

vitamin D Major building block plasma membranes 15% of cholesterol comes from diet the rest is made by the liver. Cholesterol is lost by breakdown, secretion in bile salt and finally defecation

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Lipoproteins
Fatty acids, fats, and cholesterol are insoluble in

water and therefore are transported bound to small lipid-protein complexes called lipoproteins
lipoproteins (LDL) transport cholesterol and other lipids to body cells lipoproteins (HDL) transport cholesterol from tissue cells to liver for disposal

Low-density

High-density

Ratio of HDL/LDL is important

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Body Energy Balance

When

energy intake and energy outflow are balanced body weight remains stable When they are not, weight is either lost or gained Control of food intake: ? Rising and falling blood levels of nutrients Hormones Body temperature Psychological factors

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Basal Metabolic Rate

The amount of heat produced by the body per

unit of time under basal conditions An average 155lb adult has a BMR of about 6072 kcal/hour

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Lipids Catabolism Aerobic transport of fatty acids from cytosol to

mitochondria (role of carnitine) -oxidation in mitochondria 4 steps release of NADH and FADH2 108 ATP/palmitic acid or 7 ATP/Carbon

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 Catabolism: dietary lipids Digestion:

a) Slow relative to carbohydrates b) In small intestine with action of bile salts c) FAcs absorbed across intestinal wall and reconverted to TAGs d) Transported as chylomicrons Mobilisation from adipocytes: a) FAcs transported in blood bound to serum albumin; dissociates in cells oxidation b) glycerol undergoes glycolysis glucose levels (in the liver, not the muscles)

Role of glucose 6-phosphatase in maintaining blood

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Synthesis: lipogenesis

carried out by two cytosolic enzymes, acetyl-CoA carboxylase and fatty acid synthase

Requires: NADPH, ATP and biotin, CO2 Sources of Acetyl CoA - transfer of citrate from mitoch. to
cytosol NADPH - malic enzyme in cytosol - pentose phosphate pathway

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Lipid Digestion - Rumen

-galactosidase

DigalDigly Galactose Propionate Glycerol Triglyeride

Lipase

MonogalDigly
-galactosidase

Diglyceride
Lipase Anaerovibrio lipolytica

Fatty acids
H+ Reductases

CaFA

Saturated FA Ca++

Feed particles

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Fat Digestion
Digestibility influenced by: Dry matter intake Decreases with greater intake Amount of fat consumed Digestibility decreases 2.2% for each 100 g of FA intake (Response is variable) Degree of saturation Digestibility decreases with increased saturation Maximal digestion with fats having Iodine values greater than 40
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Lipid Metabolism - In the Rumen

1. Minimal degradation of long-chain fatty acids in the rumen Fatty acids not a source of energy to microbes 2. Active hydrogenation of unsaturated fatty acids 3. Microbial synthesis of long-chain fatty acids in the rumen (15g/kg nonfat org matter fermented) 4. No absorption of long chain fatty acids from the rumen More fat leaves the rumen than consumed by the animal Lipids leaving the rumen 80 to 90% free fatty acids attached to feed particles and microbes ~10% microbial phospholipids leave the rumen Small quantity of undigested fats in feed residue
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Microbial Fatty Acid Synthesis

Synthesize C 18:0 and C 16:0 in 2:1 ratio using acetate and glucose (straight-chain even carbon #). If propionate or valerate used, straight-chain odd carbon fatty acids synthesized. Branched-chain VFA used to produce branched chain fatty acids. About 15 to 20% of microbial fatty acids are monounsaturated. No polyunsaturated fatty acids are synthesized. Some incorporation of C 18:2 into microbial lipids.
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Hydrogenation of Fatty Acids in the Rumen

Polyunsaturated fatty acids (all cis)

Isomerase (from bacteria)
Needs free carboxyl group and diene double bond

Shift of one double bond (cis & trans)

Hydrogenation
Hydrases (from bacteria,

Hydrogenated fatty acid

(stearic and palmitate)

mostly cellulolytic)

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Hydrogenation of Fatty Acids in the Rumen

All unsaturated fatty acids can be hydrogenated Monounsaturated less than polyunsaturated 65 to 96% hydrogenation Numerous isomers are produced Biohydrogenation is greater when high forage diets fed Linoleic acid depresses hydrogenation of FA

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Conjugated Linoleic Acid - Rumen Most Common Pathway (High Roughage)

Linoleic acid (cis-9, cis-12-18:2)

Cis-9, trans-12 isomerase Butyrivibrio fibrosolvens

Conjugated linoleic acid (CLA, cis-9, trans-1118:2)

Vaccenic acid (Trans-11-18:1) Stearic acid (18:0)
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CLA Isomers - Rumen (High Concentrate) Low Rumen pH

Linoleic acid (cis-9, cis-12-18:2)

Cis-9, trans-10 isomerase

CLA Isomer (trans-10, Cis-12-18:2)

This isomer is inhibitory to milk fat synthesis.

Trans-10-18:1
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Linolenic Acid Oleic Acid Linolenic acid (cis-9, cis-12, cis-15-18:3) (Cis-9, trans-11, cis-15-18:3) Trans-11, cis-15-18:2 Trans-11-18:1 (vaccenic acid) Oleic acid cis-9 (18:1) Stearic acid (18:0)

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CLA absorbed from the intestines available for incorporation into tissue triglycerides.
Reactions from linoleic acid to vaccinic acid occur at a faster rate than from vaccinic acid to stearic acid. Therefore, vaccinic acid accumulates in the rumen and passes into intestines where it is absorbed. Quantities of vaccinic acid leaving the rumen several fold greater than CLA.

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Conversion of Vaccinic Acid to CLA

In mammary gland and adipose

Trans-11-18:1
Stearoyl CoA Desaturase 9-desaturase

CLA, cis-9, trans-11 18:2

This reaction probably major source of CLA in milk and tissues from ruminants.
Also transforms Palmitic Stearic

Palmitoleic Oleic

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Synthesis: lipogenesis

carried out by two cytosolic enzymes, acetyl-CoA carboxylase and fatty acid synthase

Requires: NADPH, ATP and biotin, CO2 Sources of Acetyl CoA - transfer of citrate from mitoch. to
cytosol NADPH - malic enzyme in cytosol - pentose phosphate pathway

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 Synthesis: lipogenesis Other roles of PPP alternative pathway for glucose metabolism production of ribose 5-phosphate (nucleotide
synthesis)

Ketone bodies arise from the overflow pathway in Location of lipid metabolism oxidation in mitoch., synthesis in cytosol
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liver; major source of energy for heart, muscle and brain (fasting and diabetes)

Summary of fatty acid metabolism in the liver

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 If excess fat is consumed, there is no mechanism

by which the body can increase its use of fat as a fuel. the only option is to accumulate the excess fat as an energy store in the body, and this process occurs at a low metabolic cost and is an extremely efficient process.

Instead, when excess fat calories are consumed,

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Aulani "Biokimia" Presentation 6