Lipids and Fatty Acids Objectives: 1. What are Lipids?

•  properties •  glycerolipids vs. isoprenoids •  glycerolipid structure •  glycerolipid nomenclature

2. Fatty acid biosynthesis

•  Cellular localization •  Substrate and its origin •  Enzymes of de novo fatty acid synthesis •  De novo fatty acid elongation (C2->C18)

References: Buchanan et al. 2000. Biochemistry and Molecular Biology of Plants. American Society of Plant

Physiologists, Rockville MD. Chapter 10.

What are lipids?

Triacylglycerol (TAG)

Vitamin D2

What are lipids?

Macromolecules defined on the basis of their physical properties:

•  Insoluble in water •  Soluble in non-polar organic solvents

Lipids are chemically heterogeneous

Vitamin D2

Originate from different biosynthetic pathways

Triacylglycerol (TAG)

What are lipids?

Macromolecules defined on the basis of their physical properties:

•  Insoluble in water •  Soluble in non-polar organic solvents

Heteropolymers consisting of different subunits (glycerolipids), or do not form polymers (isoprenoids=terpenoids)

Glycerolipids Fats and oils

H C

H C

H C

H

H

H C

H C

H C

H

H

1

2

3 H C

H C

H C

H

H

H C

H C

H C

H

H

1

2

3

Glycerol backbone

Fatty acyl group

Triacylglycerol (TAG)

sn-1

sn-2

sn-3

Triacylglycerols form oil droplets

Oilseed embryo

Glycerolipids

Acyl group

Membrane lipids

HOH

OH

H2COH

HH

H

O O H OH

H C

H

H C

H C

H

Glycerol backbone

Fatty acyl group

Head group

Monogalactosyldiacylglycerol (MGDG)

sn-1

sn-3

sn-2

Glycerolipids

Acyl group

Membrane lipids

H C

H

H C

H C

H

Glycerol backbone

Fatty acyl group

Phosphatidylcholine (PC)

sn-1

sn-3

sn-2

O

O

O

O

O

P

Head group

1. Glycerol backbone

2. Fatty acyl groups on sn-1 and sn-2 positions

•  Saturated – no double bonds •  Unsaturated – one or multiple double

bonds

3. Head group on sn-3 position

•  Sugar moiety -> Glycolipids •  Phosphate + alcohol -> Phospholipids •  No head group -> Acyl glycerols

Glycerolipid nomenclature

Oleic acid 18:1 Δ9 or ω9

Number of Carbon atoms

Number of Double bonds

Trivial name

Double bond position (counting from carboxyl end or methyl end)

cis configuration unless indicated

Fatty acid nomenclature -

H C C

H - -

H C C

H - -

H C - H C C

H C H

-

trans double bond E configuration Carboxyl end

Methyl end

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

O

HO C - C

H

- C H

- H C H

- H C H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H

H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

O

HO C - C

H

- C H

- H C H

- H C H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H

H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

O

HO C - C

H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

O

HO C - C

H C H

- C H

- H C H

- H C H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H

H

- C H - C

H

- H C H

- H C H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H

H

- H C H

- H C H -

H C H

- H C H

- H C H

- H C H -

H C H

- H C H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H -

H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

- H C H

cis double bond Z configuration

Common Eukaryotic Fatty Acids

Unsaturated fatty acids

16:0 Hexadecanoic acid Palmitic acid

18:0 Octadecanoic acid Stearic acid

16:1 Δ7 Hexadecenoic acid Palmitoleic acid

16:1 Δ3 t trans-hexadecenoic acid

16:2 Δ7Δ10 Hexadecadienoic acid

16:3 Δ7,Δ10,Δ13 Hexadecatrienoic acid 18:1 Δ9 Octadecenoic acid Oleic acid

18:2 Δ9,Δ12 Octadecadienoic acid Linoleic acid

18:3 Δ9,Δ12,Δ15 Octadecatrienoic acid Linolenic acid

Saturated fatty acids

Symbol Systematic name Trivial name

•  16 or 18-carbon unbranched acyl chain (even number)

Characteristics of Common Fatty Acids

•  Zero to 3 double bonds

•  Double bonds are methylene interrupted (Allylic)

=CH

-CH

-HCH

-CH

-HCH

-HCH

=CH

- =CH

-CH

-HCH-C

H-

HCH

-CH

-HCH

-HCH

=CH

-

Allylic CH2 group

•  Components of membrane and storage lipids

De novo Fatty Acid Synthesis

Fig. 10-9

Site: PLASTID Substrate: acetyl-CoA

De novo Fatty Acid Synthesis

Fig. 10-11

Acetyl-CoA Carboxylase reaction 1st committed step in fatty acid synthesis

FAS Enzymes

ACP = Acyl Carrier Protein – primer for FAS, carries the fatty acyl chain

6. ER = 2-3 trans enoyl-ACP reductase

Enzymes:

1. ACCase = Acetyl-CoA Carboxylase

3. CONDENSING ENZYMES – key enzymes of FAS involved in C-C bond formation (CO2 release – IRREVERSIBLE!)

•  KAS III – C2 •  KAS I – C4-C14 •  KAS II – C16

– different substrate specificities:

4. KR = 3-ketoacyl-ACP reductase

5. DH = 3-hydroxyacyl-ACP dehydratase

2. Malonyl-CoA:ACP transacylase

Organization of FAS Enzymes

TYPE I (Animals and Fungi)

TYPE II (Plants and Bacteria)

Location: Cytosol

Location: Plastid stroma

Organization: A complex of 2 large subunits (250 kDa each) catalyzing all

the reactions of FAS

Organization: Enzyme activities reside on individual proteins – this reflects prokaryotic origin of plastids

KAS KR

DH ER

KAS KR

DH ER

Fatty Acid Synthesis

Ohlrogge and Browse (1995) Plant Cell 7:795

KASIII

ER

DH

KR

KAS I

ACC

TA

Fatty Acid Synthesis

FAS

16:0-ACP

18:0-ACP

18:1-ACP

FATB

FATA

LACS

LACS

16:0-CoA

18:1-CoA

AT

AT

16:0

18:1 Chl

orop

last

lipi

ds

DES

FAS = fatty acid synthesis

FAT = acyl-ACP thioesterase

AT = acyl transferase

LACS = acyl-CoA synthetase

DES = stearoyl-ACP desaturase

Main products of FAS are 16:0 and 18:1

AT 18:0 18:0-CoA LACS

FATB KASII

Main products of FAS are 16:0 and 18:1

Export to the ER Lipid synthesis in the chloroplast

AT FAT

How do we know that?

From (14C)-acetate labeling experiments of isolated plastids in vitro

C16 and C18 fatty acyl chains get radioactively labeled