Chemical Properties of Amino Acids

Reactions of Amino Acids of Biological Importance

The functional groups of amino acids retain their properties and, therefore,
offer a complete variety of reactivity of both amino group and carboxyl group. Of
course, the mutual influence of these electron-withdrawing groups modifies their
reactivity at a certain level. One has to consider also that both the amino group and
the carboxyl group can be protonated and deprotonated, therefore, their reactivity
depends on acidity (pH) of the reaction medium.

Amino acids can produce salts:

The carboxyl group of the amino acids can react with alcohols to produce
esters. Acidic medium is mandatory:

The amino group can also be acylated by acid anhydrides or acyl halides:

The amino group can also react with aldehydes and ketones. The products of
this reaction are imines (Schiff’s bases):
 Heating of α-amino acids produces cyclic diamides, diketopyperazines:

heating 2
2

γ- and δ-amino acids, however, produce cyclic monoamides, γ- and δ-lactams

correspondingly:

Heating of β-amino acids leads to formation of α,β-unsaturated carboxylic

acids:
 ε-amino acids and some other amino acids where the amino group is placed
even farther from the carboxyl group produce linear polyamides via heating:

As we see, heating of amino acids allows for differentiation among various

types of amino acids and is fully analogical to heating of hydroxy acids.

However, the most important reactions of α-amino acids involve

decarboxylation, deamination and transamination.

Decarboxylation of α-amino acids produces amines. This process is guided

with Ba(OH)2 in vitro (heating is required), whereas pyridoxal phosphate as a
coenzyme is required for in vivo conversions:

decarboxylases (enzymes)

-CO2

As we have seen, the carboxyl group of the side chain does not undergo
decarboxylation in these conditions. Tryptamine, histamine and GABA are among
most important products of decarboxylation of α-amino acids.
 There are four types of deamination of α-amino acids. Hydrolytic,
intramolecular, and reducing deamination are available only in vitro, but oxidative
(oxidation) deamination can also be an in vivo process.

Reducing deamination produces carboxylic acids:

2[H]

-NH3
Hydrolytic deamination produces α-hydroxy acids:

+H2O

-NH3

Intramolecular deamination produces α, β-unsaturated carboxylic acids:

-NH3

And oxidative (oxidation) deamination produces α-keto acids:

 [O]

-NH3

Responsible for the in vivo oxidation deamination is pyridoxal phosphate that

serves as a coenzyme. It is bound to the amino group of the side chain of lysine
residue and, therefore, presents itself as a Schiff’s base. A similar mechanism is
employed for transamination. An overall net reaction can be presented as follows:

As am example, let us consider transamination of aspartic acid with α-

ketoglutaric acid with aspartate aminotransferase as enzyme:

Transamination first occurs when aspartic acid (amino acid 1) takes pyridoxal
phosphate from the lysine residue. The resultant imine is then hydrolyzed to keto
acid (keto acid 2) and pyridoxamine phosphate; the latter reacts with α-ketoglutaric
acid (keto acid 1). The resultant imine is once again transaminated with the lysine
residue and the target amino acid (amino acid 2, here glutamic acid) is released.

It is important to remember that lysine, tyrosine and proline normally do not

undergo transamination. Let us also keep in mind that, while theoretically any α-keto
acid can serve as a vehicle for the amino group transfer, these are only pyruvic acid,
oxaloacetic acid and α-ketoglutaric acid that are discovered participating in
transamination in vivo.

The same coenzyme, pyridoxal phosphate (PLP), that is a derivative of

vitamin B6, is also responsible for racemization of α-amino acids leading to
formation of D-amino acids.

Analytically Important Reactions

Some reactions of amino acids are important for detection and measurement
of amino acid content. We can group these reactions into two classes. The first
group if reactions is based on properties of the main fragment of α-amino acids. The
second group consists of reactions that are specific to side chains of amino acids. The
typology of the side chains is versatile and so are the reactions aiming at their
detection.

The Van Slyke’s method is based in hydrolytic deamination of amino acids.

When α-amino acids react with nitrite salts, molecular gaseous nitrogen is released.
By measurement of the latter, one can calculate the content of primary amino
groups:
 This reaction carries out only for primary amines as two hydrogen atoms at
the nitrogen atom of the amino group are required for nitrosamine formation and its
tautomeric conversion to the oxime form (diazotic acid):

Sorensen’s test is based on protection of the amino group with formaldehyde.

Protons are released and their concentration is measured by titration, which allows
for calculation of the initial concentration of the α-amino acid in question:
 UV-spectroscopic analysis of α-amino acids employs their reaction with
ninhydrin. A highly conjugated molecule is produced after series of imine formation,
decarboxylation, hydrolysis, and imine formation again. The resultant molecule is
such that the structure of the amino acid in question is not reflected:

The absorption maximum in the UV-spectrum is ~570 nm.

The product of the reaction of proline (that is a secondary amine) produces

yellowish product; its structure is different from what we have seen above as
secondary amines produces enamines, not imines, via reactions with carbonyl
compounds.
 Certain amino acids can be identified with specific reactions of their side
chains. Tryptophan can be detected with p-dimethylaminobenzaldehyde in presence
of sulfuric acid. The solution of the product acquires reddish-violet color. Cysteine is
a thiol and can be detected with lead acetate. Amino acids containing aromatic rings
in the side chain can be identified by their reaction with concentrated nitric acid and
the resultant yellow color.