Lipids

Lipids (Greek: lipos–fat) are of great importance to the body as the chief
concentrated storage form of energy, besides their role in cellular
structure and various biochemical functions.
Lipids can be regarded as heterogeneous group of organic compounds,
relatively insoluble in water, soluble in organic solvents (alcohol, ether
etc.), related to fatty acids and utilized by the living cells. Unlike the
polysaccharides, proteins, and nucleic acids, lipids are not polymers.
Further, lipids are mostly small molecules.

Classification of lipids
Lipids are broadly classified into simple, complex, derived, and
miscellaneous lipids, which are further subdivided into different groups.
1. Simple lipids: Esters of fatty acids with alcohols. These are mainly of
two types; 1
a. Fats and oils (triacylglycerols): These are esters of fatty acids with
glycerol. Oil is a liquid while fat is a solid at room temperature.
b. Waxes: Esters of long chain fatty acids with alcohols, other than
glycerol. These alcohols may be aliphatic or alicyclic. Cetyl alcohol is
most commonly found in waxes. Waxes are used in the preparation of
candles, lubricants, cosmetics, ointments, polishes etc.
2. Complex (compound) lipids: These are esters of fatty acids with
alcohols containing additional groups, like phosphate, nitrogenous
base, carbohydrate, and protein. They are further divided as,
a. Phospholipids: They contain phosphoric acid and frequently a
nitrogenous base in addition to alcohol and fatty acids.
i. Glycerophospholipids: These phospholipids contain glycerol as the
alcohol e.g., lecithin, cephalin.
ii. Sphingophospholipids: Sphingosine is the alcohol in this group of
phospholipids e.g., sphingomyelin. 2
b. Glycolipids (glycosphingolipids): These lipids contain a fatty acid,
carbohydrate, and nitrogenous base. The alcohol is sphingosine.
Glycerol and phosphate are absent. e.g., cerebrosides, gangliosides.
c. Lipoproteins and Lipopolysaccharides: Macromolecular complexes
of lipids with proteins and polysaccharides respectively.
d. Other complex lipids: e.g., Sulfolipids, and aminolipids.
3. Derived lipids: These are the derivatives obtained on the hydrolysis
of simple and complex lipids. These include glycerol and other
alcohols, fatty acids, mono-and diacylglycerols, fat soluble vitamins,
steroid hormones, hydrocarbons, and ketone bodies.
4. Miscellaneous lipids: These include a large number of compounds
possessing the characteristics of lipids. e.g., carotenoids, squalene,
pentacosane (in bees wax), terpenes etc.
3
Neutral lipids
The uncharged lipids are referred to as neutral lipids. e. g.., These are
mono- , di-, and triacylglycerols, cholesterol, cholesteryl esters.
Functions of lipids
Lipids perform several important functions;
1. Triglycerides, serve as an efficient energy reservoir of the body
When the body needs energy, it breaks down stored triglycerides
into fatty acids, releasing energy for various cellular processes.
2. Lipids are the constituents of membrane structure and regulate the
membrane permeability. Phospholipids are the main lipid
constituents of membrane. Cholesterol (steroid and an alcohol)
provides a rigid hydrophobic structure of the cell membrane.
3. They serve as a source of fat soluble vitamins (A, D, E, and K).
4. Waxes, contribute to the structural integrity of certain organisms. Eg.
Plant waxes help to prevent water loss and protect leaves. 4
5. Lipids cushion and protect the internal organs, acting as a natural
shock absorber. Adipose tissue, composed mainly of lipids, acts as an
insulating layer beneath the skin and provide shape, and smooth
appearance to the body. It helps regulate body temperature by
minimizing heat loss.
6. Lipids are important as cellular metabolic regulators (Eg. steroid
hormones, and prostaglandins). Steroids, a lipid subclass, play a key
role in hormone synthesis. Hormones such as estrogen, testosterone,
and cortisol are derived from cholesterol (a type of steroid).
7. Some lipids, particularly steroids, act as chemical messengers within
the body. They regulate various physiological processes, including
growth, metabolism, and immune responses.
8. Glycolipids, which are part of cellular lipid pools, play roles in cell
recognition and signaling.
In summary, lipids are not only essential for energy storage but also
5
significant in the functionality and survival of living organisms.
 Fatty acids
Fatty acids are carboxylic acids with hydrocarbon side chain. They are
the simplest form of lipids. They occur in the esterified form as major
constituents of various lipids. They are also present as free
(unesterified) fatty acids.

Even and odd carbon fatty acids: Most of the fatty acids that occur in
natural lipids are of even carbons (usually 14C – 20C). This is due to the
fact that biosynthesis of fatty acids mainly occurs with the sequential
addition of 2 carbon units. Palmitic acid (16C) and stearic acid (18C) are
the most common. Among the odd chain fatty acids, propionic acid (3C)
and valeric acid (5C) are well known.

6
Saturated and unsaturated fatty acids
Saturated fatty acids do not contain double bonds, while unsaturated
fatty acids contain one or more double bonds. Both saturated and
unsaturated fatty acids equally occur in the natural lipids. Fatty acids
with one double bond are monounsaturated, and those with 2 or more
double bonds are collectively known as polyunsaturated fatty acids
(PUFA).

Nomenclature of fatty acids

The naming of a fatty acid (systematic name) is based on the
hydrocarbon from which it is derived. The saturated fatty acids end with
a suffix -anoic (e.g., octanoic acid [C8]) while the unsaturated fatty acids
end with a suffix -enoic (e.g., octadecenoic acid [18C]). In addition to
systematic names, fatty acids have common names which are more
widely used (Table 3.1). 7
Table 3.1:
Selected
examples of
biochemically
important
fatty acids.

8
Numbering of carbon atoms
It starts from the carboxyl carbon (number 1) and the carbons adjacent
to this (carboxyl C) are 2, 3, 4 and so on or alternately α, β, γ and so on.
The terminal carbon containing methyl group is known as omega (ω)
carbon. Starting from the methyl end, the carbon atoms in a fatty acid
are numbered as omega 1, 2, 3 etc. The numbering of carbon atoms in
two different ways is given below.

Heptanoic acid - CH3(CH2)5COOH

Length of hydrocarbon chain of fatty acids
Depending on the length of carbon chains, fatty acids are categorized
into 3 groups—short chain with <6 carbons; medium chain of 6-12
carbons and long chain of 12-24 carbons. 9
Shorthand representation of fatty acids
Instead of writing the full structures, biochemists employ shorthand
notations (by numbers) to represent fatty acids. The general rule is that
the total number of carbon atoms are written first, followed by the
number of double bonds and finally, the first carbon position of double
bonds, starting from the carboxyl end. Thus, saturated fatty acid,
palmitic acid is written as 16 : 0; oleic acid as 18 : 1; 9; arachidonic acid
as 20 : 4; 5, 8, 11, 14.
There are other conventions of representing the double bonds. Δ9
indicates that the double bond is between 9 and 10 of the fatty acid. ω9
represents the double bond is between 9 and 10 from the ω end.
Naturally occurring unsaturated fatty acids belong to ω3, ω6, and ω9,
series.

10
Essential fatty acids (EFA)
The fatty acids that cannot be synthesized by the body and should be
supplied in the diet are known as EFA. Chemically, they are
polyunsaturated fatty acids, namely linoleic acid (18 : 2; 9, 12) and
linolenic acid (18 : 3; 9, 12, 15). Arachidonic acid (20 : 4; 5, 8, 11, 14)
becomes essential, if its precursor linoleic acid is not provided in the diet.
The structures of EFA are given in the Table 3.1. Linoleic acid and linolenic
acid are essential since humans lack the enzymes that can introduce
double bonds beyond carbons 9 - 10.

Functions and Deficiencies of EFA

Essential fatty acids are required for the membrane structure and
function, transport of cholesterol, formation of lipoproteins, prevention
of fatty liver, and synthesis of eicosanoids. The deficiency of EFA results in
phrynoderma or toad skin (horny eruptions). 11
Isomerism in unsaturated fatty acids
Unsaturated fatty acids exhibit geometric isomerism depending on the
orientation of the groups around the double bond axis. If the atoms/
groups are present on the same side of the double bond, it is a cis
configuration. On the other hand, if the atoms/ groups occur on the
opposite side, it is a trans configuration. Thus oleic acid is a cis isomer
while elaidic acid is a trans isomer, as depicted in Fig.3.1. Cis isomers
are less stable than trans isomers. Most of the naturally occurring
unsaturated fatty acids exist as cis- isomers.

12
In the cis- isomeric form, there is a molecular bend at the double bond.
Thus, oleic acid exists in an L-shape, while elaidic acid is a straight chain.
Arachidonic acid with 4 double bonds will have a U-shape. It is believed
that cis- isomers of fatty acids with their characteristic bends will
compactly pack the membrane structure.

Triacylglycerols (fats and oils)

Triacylglycerols (formerly triglycerides) are the esters of glycerol with
fatty acids. The fats and oils of plants and animals are chemically
triacylglycerols. They are insoluble in water and nonpolar in character
and commonly known as neutral fats. Triacylglycerols are the most
abundant group of lipids that primarily function as fuel reserves of
animals in adipose tissue and as globules dispersed in cytoplasm. And
surprisingly, triacylglycerols are not the structural components of
biological membranes. 13
Structures of acylglycerols
Monoacylglycerols, diacylglycerols and triacylglycerols, respectively
consisting of 1, 2, and 3 molecules of fatty acids esterified to a molecule
of glycerol are known as acylglycerols. Among these, triacylglycerols are
the most important biochemically.

14
Simple triacylglycerols contain the same type of fatty acid residue at all
the 3 carbons e.g., tristearoyl glycerol or tristearin.
Mixed triacylglycerols contain 2 or 3 different types of fatty acid
residues. In general, fatty acid attached to C1 is saturated, that attached
to C2 is unsaturated while that on C3 can be either.
Triacylglycerols are named according to placement of acyl radical on
glycerol. e.g., 1,3-palmitoyl 2- linoleoyl glycerol.
Triacylglycerols of plants have higher content of unsaturated fatty acids
compared to that of animals.
Stereospecific numbering (sn) represent the carbon
atoms (1, 2, and 3) of glycerol in an unambiguous
manner as 1 and 3 carbon atoms are different in 3-
dimensional structure and prefix glycerol with sn.
Glycerokinase in cell phosphorylates sn-3 (not sn-1)
glycerol to give glycerol 3-phosphate. 15
Properties of triacylglycerols
1.Hydrolysis: Triacylglycerols undergo stepwise enzymatic hydrolysis to
liberate free fatty acids and glycerol. The process of hydrolysis, catalysed
by lipases is important for digestion of fat.
2.Saponification: The hydrolysis of triacylglycerols by alkali to produce
glycerol and soaps is known as saponification.

3.Rancidity: Rancidity represents the deterioration of fats and oils resulting

in an unpleasant taste. Fats and oils containing unsaturated fatty acids
are more susceptible to rancidity. It occurs when they are exposed to air,
moisture, light, bacteria etc. Hydrolytic rancidity occurs due to partial
hydrolysis of triacylglycerols by bacterial enzymes. Oxidative rancidity is
due to oxidation of unsaturated fatty acids. This results in the formation
of unpleasant products such as dicarboxylic acids, aldehydes, ketones
etc. Rancid fats and oils are unsuitable for human consumption. 16
Tests to check purity (adulteration) of fats and oils
1. Iodine number is defined as the number of
grams of iodine absorbed by 100 g of fat or oil.
Iodine number is directly proportional to the
content of unsaturated fatty acids. Thus, lower is
the iodine number, lesser the degree of
unsaturation. Determination of iodine number will
help to know the degree of adulteration of oil.
The iodine numbers
2. Saponification number is defined as the number of common oils/ fats
mg’s of KOH required to hydrolyze (saponify) 1 g of
fat or oil. Saponification number is a measure of
the average molecular size of the fatty acids
present. The value is higher for fats containing
The saponification
short chain fatty acids. numbers of fats/ oils 17
 Proteins
The term protein (Greek: proteios), meaning holding the first place.
Berzelius (Swedish chemist) suggested the name proteins to the group of
organic compounds that are utmost important to life. Mulder (Dutch
chemist) in 1838 used the term proteins for high molecular weight
nitrogen rich molecules in animals and plants. Proteins occur in every
part of the cell and constitute about 50% of the cellular dry weight.
Proteins form the fundamental basis of structure and function of life.

Functions: Proteins perform a great variety of specialized and essential

functions in the living cells.
1. Structural functions: Certain proteins are primarily responsible for
structure and strength of body. These include collagen and elastin
found in bone matrix, vascular system and other organs, and α-
keratin present in epidermal tissues. 18
2. Dynamic functions: This include proteins acting as enzymes,
hormones, blood clotting factors, immunoglobulins, membrane
receptors, storage proteins, besides their function in genetic control,
muscle contraction, respiration etc. Proteins performing dynamic
functions are appropriately regarded as the working horses of cell.

Elemental composition of proteins

Proteins are predominantly constituted by 5 major elements in the
following proportion. Carbon: 50 – 55%; Hydrogen: 6 – 7.3%; Oxygen: 19
– 24%; Nitrogen: 13 – 19%; and Sulfur: 0 – 4%. Proteins may also contain
other elements such as P, Fe, Cu, I, Mg, Mn, Zn etc.

Proteins are polymers of amino acids

Proteins are polymers of L-α-amino acids as complete hydrolysis of
proteins with conc. HCl for several hours yield L-α-amino acids. 19
Standard amino acids
As many as 300 amino acids occur in nature, of these, only 20 known as
standard amino acids are repeatedly found in the structure of proteins
isolated from animal, plant, and microbe. This is because of the universal
nature of the genetic code available for the incorporation of only 20
amino acids when proteins are synthesized.

Amino acids
Amino acids are organic compounds containing two functional groups;
amino and carboxyl. The amino group (—NH2) is basic, while the
carboxyl group (—COOH) is acidic in nature.

General structure of amino acids

The amino acids are termed as α-amino acids, if both the carboxyl and
amino groups are attached to the same carbon atom. 20
The α-carbon atom binds to a side chain represented by R which is
different for each of the 20 amino acids. The amino acids mostly exist as
ionized form in the biological system.
Optical isomers of amino acids
If a carbon atom is attached to four different groups, it is asymmetric
and therefore exhibits optical isomerism. The amino acids (except
glycine) possess four distinct groups held by α-carbon. Thus all the
amino acids (except glycine where R is ‘H’) have optical isomers. The
structures of L-and D-amino acids are written based on the
configuration of L-and D-glyceraldehyde as shown in Fig.4.1. The
proteins are composed of L-α-amino acids. 21
FIG. 4.1. D-and L-forms of amino acid based
on the structure of glyceraldehyde.

Classification of amino acids

The different ways of classifying amino acids are based on the structure
and chemical nature, nutritional requirement, metabolic fate etc.
1. Based on the structure and chemical nature
Each amino acid is assigned a 3 letter or 1 letter symbol. These
symbols are commonly used to represent the amino acids in protein
structure. The 20 amino acids found in proteins are divided into 7
distinct groups. In Table 4.1, the different groups of amino acids, their
symbols and structures are given. 22
1. Amino acids with aliphatic side chains: These are monoamino
monocarboxylic acids. This group consists of simple amino acids
glycine, alanine, valine, leucine, and isoleucine. The last three amino
acids (Val, Leu, Ile,) contain branched aliphatic side chains, hence
they are referred to as branched chain amino acids.
2. Hydroxyl group containing amino acids: Serine, threonine, and
tyrosine are hydroxyl group containing amino acids. Tyrosine being
aromatic in nature, is considered under aromatic amino acids.
3. Sulfur containing amino acids: Cysteine with sulfhydryl group and
methionine with thioether group are the two amino acids
incorporated during the course of protein synthesis. Cystine, another
important sulfur containing amino acid, is formed by condensation
of 2 molecules of cysteine.
23
Table 4.1
Structural
classification
of L-α-amino
acids found in
proteins

24
25
4. Acidic amino acids and their amides: Aspartic acid and glutamic
acids are dicarboxylic monoamino acids while asparagine and
glutamine are their respective amide (R−C(=O)−NR ′R″) derivatives.
They are highly acidic in character and all these four amino acids
possess distinct codons for their incorporation into proteins.
5. Basic amino acids: Lysine, arginine (with guanidino group; )
and histidine (with imidazole ring) are dibasic monocarboxylic acids.
They are highly basic in character.
6. Aromatic amino acids: Phenylalanine, tyrosine, and tryptophan
(with indole ring) are aromatic amino acids.
7. Imino acids: Proline containing pyrrolidine ring is a unique amino
acid. It has an imino group (=NH), instead of an amino group (–NH2)
found in other amino acids. Therefore, proline is an α-imino acid.
26
27
28
Heterocyclic amino acids: Histidine, tryptophan, and proline.
B. Classification of amino acids based on polarity: Polarity of amino
acids is important for protein structure.
1. Non-polar amino acids: These amino acids are hydrophobic. They
have no charge on the ‘R’ group. The amino acids included in this
group are alanine, valine, leucine, isoleucine, methionine,
phenylalanine, tryptophan, and proline.
2. Polar amino acids: These amino acids are hydrophilic. These amino
acids, as such carry no charge on the ‘R’ group. However, they
possess groups such as hydroxyl, sulfhydryl, and amide and
participate in hydrogen bonding of protein structure. The amino
acids in this group are glycine, serine, threonine, cysteine,
asparagine, glutamine, and tyrosine.
29
3. Polar amino acids with positive ‘R’ group: The 3 amino acids lysine,
arginine, and histidine are included in this group.
4. Polar amino acids with negative ‘R’ group: The dicarboxylic
monoamino acids such as aspartic acid and glutamic acid.

C. Nutritional classification of amino acids: The amino acids are

required for the synthesis of variety proteins, besides other
biological functions. However, all these 20 amino acids need not be
taken in the diet. Based on the nutritional requirements, amino acids
are grouped into two classes—essential and nonessential.
1. Essential or indispensable amino acids: The amino acids which
cannot be synthesized by the body and, therefore, need to be
supplied through the diet are called essential amino acids. They are
required for proper growth and maintenance of the individual.
30
 Ten amino acids are essential for humans: Arginine, Valine, Histidine,
Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Threonine,
Tryptophan [code A.V. HILL, MP., T. T]. Amino acids namely arginine
and histidine can be synthesized by adults and not by growing
children, hence these are considered as semi–essential amino acids
(AH). Thus, 8 amino acids are essential while 2 are semi-essential.
2. Nonessential or dispensable amino acids: The body synthesize about
10 amino acids to meet the biological needs, hence they need not be
consumed in the diet. These are glycine, alanine, serine, cysteine,
aspartate, asparagine, glutamate, glutamine, tyrosine, and proline.
D. Amino acid classification based on their metabolic fate: The carbon
skeleton of amino acids can serve as a precursor for the synthesis of
glucose (glycogenic) or fat (ketogenic) or both.
31
1. Glycogenic amino acids: These amino acids can serve as precursors
for the formation of glucose or glycogen. e.g. alanine, aspartate,
glycine, and methionine.
2. Ketogenic amino acids: Fat can be synthesized from these amino
acids. Leucine and lysine are exclusively ketogenic.
3. Glycogenic and ketogenic amino acids: Isoleucine, phenylalanine,
tryptophan, tyrosine are precursors for synthesis of glucose as well
as fat.
Selenocysteine: The 21st amino acid found at the active sites of
certain enzymes or selenoproteins. e.g. glutathione peroxidase,
glycine reductase, 5′-deiodinase, thioredoxin reductase.
Selenocysteine is an unusual amino acid containing the trace element
selenium in place of the sulfur atom of cysteine. The stop codon UGA
code selenocysteine.
Pyrrolysine: The 22nd amino acid present in protein. The stop codon
UAG code for pyrrolysine. 32
Properties of amino acids
The amino acids differ in their physio–chemical properties which
ultimately determine the characteristics of proteins.
A. Physical properties
1. Solubility: Most of the amino acids are usually soluble in water and
insoluble in organic solvents.
2. Melting points: Amino acids generally melt at higher temperatures,
often above 200 °C.
3. Taste: Amino acids may be sweet (Gly, Ala, Val), tasteless (Leu) or
bitter (Arg, Ile). Monosodium glutamate (ajinomoto) is used as a
flavoring agent in food industry to increase taste and flavor.
4. Optical properties: All the amino acids except glycine possess optical
isomers due to the presence of asymmetric carbon atom. Some
amino acids have a second asymmetric carbon. e.g. isoleucine,
threonine. 33
5. Amino acids as ampholytes: Amino acids contain both acidic
(–COOH) and basic (–NH2 ) groups. They can donate a proton or
accept a proton, hence amino acids are regarded as ampholytes.

Zwitterion (dipolar ion)

Zwitter ion (dipolar ion) is a hybrid molecule containing positive and
negative ionic groups. The amino acids rarely exist in a neutral form
with free carboxylic (–COOH) and free amino (–NH2 ) groups. In strongly
acidic pH (low pH), the amino acid is positively charged (cation) while in
strongly alkaline pH (high pH), it is negatively charged (anion). Each
amino acid has a characteristic pH (PI) (e.g. leucine, pH 6.0) at which it
carries both positive and negative charges and exists as zwitterion
(Fig.4.2).
34
Isoelectric pH (pI) is defined as the pH
at which a molecule exists as a
zwitterion ion and carries no net
charge. Thus, the molecule is
electrically neutral.

The pI value can be calculated by

taking the average pKa values
corresponding to the ionizable
groups. For instance, leucine has two
ionizable groups, and its pI can be
calculated as follows;
FIG. 4.2 Existence of an amino
acid as cation, anion and
zwitterion.
35
Leucine exists as cation at pH below 6 and anion at pH above 6. At the
isoelectric pH (pI=6.0), leucine is found as zwitterion. Thus the pH of
the medium determines the ionic nature of amino acids. For the
calculation of pI of amino acids with more than two ionizable groups,
the pKa’s for all the groups have to be taken into account.

B. Chemical properties
The general reactions of amino acids are mostly due to the presence of
two functional groups namely carboxyl (–COOH) group and amino
(–NH2) group.
Reactions due to –COOH group
1. Amino acids form salts (–COONa) with bases and esters (–COOR′)
with alcohols.
2. Decarboxylation: Amino acids undergo decarboxylation to produce
corresponding amines. 36
Decarboxylation reaction assumes significance in the living cells due to
the formation of many biologically important amines. These include
histamine, tyramine and ϒ-amino butyric acid from the amino acids
histidine, tyrosine, and glutamate, respectively.
3. Reaction with ammonia: The carboxyl group of dicarboxylic amino
acids reacts with NH3 to form amide

4. Salt formation (reaction of –NH2 group): The amino groups behave

as bases and combine with acids (e.g. HCl) to form salts (-NH3+Cl-).
37
5. Reaction with ninhydrin: The α-amino carboxylic acids react with
ninhydrin to form a purple, blue, or pink colour complex
(Ruhemann's purple). Ninhydrin reaction is effectively used for the
quantitative determination of amino acids and proteins. (Note:
Proline and hydroxyproline give yellow colour with ninhydrin).
6. Colour reactions: Amino acids can be identified by specific colour
reactions (Table 4.3).
Table 4.3. Colour reactions of proteins/amino acids

38
7. Transamination: Transfer of an amino group from an amino acid to a
keto acid to form a new amino acid is a very important reaction in
amino acid metabolism.

8. Oxidative deamination: The amino acids undergo oxidative

deamination to liberate free ammonia. Simultaneously, oxidative
deamination produces α- keto acids (such as α- ketoglutarate). These
α- keto acids serve as intermediates for various reactions, including
energy generation.
39