Index

1. Introduction
2. Definition
3. Structure
4. Classification
5. Distribution
6. Physical Properties
7. Biosynthesis: Biochemistry
 Biochemistry
 Biosynthesis Regulation
8. Functions
 Taxonomic Markers
 Ecological and Physiological Aspects
 Pharmacological Effects
9. Methodological aspects
 Extraction
 Seperation
10. Chemical Properties
 Stability in Model Systems
 Processing and Stability in Foods
 Production of Betalains by Plant Tissue Culture
11.Bibliography
Betalaines
Table beets are a good source of red pigments; these have been increasingly used
for food coloring. Plants containing betalains have colors similar to plants containing
anthocyanins. Betalains are a group of pigments containing betacyanins (red) and
betaxanthins (yellow) and their color is not affected by pH, contrary to the behavior
of anthocyanins. The red and yellow pigments obtained from beets are known as
betalains and consist of the red betacyanins and the yellow betaxanthins.
Betalaines are water soluble and exist as internal salts (zwitterions) in the vacuoles
of plant cells. Plants containing these pigments are restricted to ten families of the
order Centrospermae.

The major betacyanin is betanin, which accounts for 75–95% of the total pigments
of beets. The remaining pigments contain isobetanin, prebetanin, and
isoprebetanin. The latter two are sulfate monoesters of betanin and isobetanin,
respectively. The major yellow pigments are vulgaxanthin I and vulgaxanthin. II
Betanin is the glucoside of betanidin, and isobetanin is the C-15 epimer of betanin.
Definition

The term “betalains” was introduced by Mabry and Dreiding; this was supported by
structural and biogenetic considerations. In a strict sense, betalains do not belong
to alkaloids because they are acidic in nature due to the presence of several
carboxyl groups. Originally, betalains were called “caryophyllinenroth” and
successively renamed “rübenroth” and “chromoalkaloids”.

Structure

Chemically, betalain definition embraces all compounds with structures based on

the general formula:

The general formula for betalaines represents condensation of a primary or

secondary amine with betalamic acid therefore, they are ammonium derivatives of
betalamic acid.

The ch romoph ore of betal ain s can be describ ed a s a p r o t o n a t e d

1 , 2 , 4 , 7 , 7 p e n t a s u b s t i t u e d 1,7-diazaheptamethin system. When R’ does not
extend conjugation of the 1,7-diazaheptamethine system, the compound exhibits
maximum light absorption at about 480 nm, characteristic of yellow betaxanthins. If
the conjugation is extended at R’ the maximum light absorption shifts to
approximately 540 nm, characteristic of red betacyanins.
Betacyanin structures show some variations in the acyl groups and sugar
moieties,while betaxanthin exhibits the same dihydropyridine moiety but show
conjugation with several amines and amino acids. Betacyanins are optically active
because of the two chiral carbons C-2 and C-15. Hydrolysis of betacyanin leads to
either betanidin, or the C-15 epimer isobetanidine, or a mixture of the two isomeric
aglycones. These aglycones are shared by all betacyanines. Differences between
betacyanins are found in their glucoside residue. Common vegetables containing
betalaines are red beet and amaranth. The latter is either consumer fresh as
“greens” or at the mature state as grain. The most studied betalaines are those of
the red beet. The major betacyanins in the red beet are betanin and isobetanin,
while in amaranth they are amarathin and isoamaranthin.

Betaxanthins are constituted of different proteinogenic and nonproteinogenic amino

acids, as well as biogenic amine-conjugated moieties of betalamic acids. More than
200 amino acids found in plants may potentially give rise to betaxanthin structures.
The first betaxanthin isolated and characterized was indicaxanthin, isolated from
prickly pear (cactus fruits of Opuntia ficus-indica). Structurally these pigments are
very similar to betacyanins. Betaxathins differ from betacyanins in that the indole
nucleus is replaced with an amino acid. In the case of indicaxanthin the amino acid
is proline. Two betaxanthines have been isolated from beet, vulgaxanthin I and II.
They differ from indicaxanthine in that the proline has been replaced by glutamine
or glutamic acid, respectively. Although only a few betaxanthines have been
characterized to date, considering the number of amino acids available, it is likely a
large number of different betaxanthins exist.

Betanidin is the basic structural unit of most of the betacyanins, followed by its C 15
epimer, the isobetanidin. Betanidin has three carboxyl groups (pKa = 3.4), two
phenol groups (pHa = 8.5), and asymmetric carbons at positions 2 and 15. The 15-
position is easily isomerized under acid or basic conditions in the absence of oxygen
to yield isobetanidin. Under alkaline conditions and in the presence of glutamine or
glutamic acid, betanin can be converted to vulgaxanthin. A considerable number of
different betacyanins can be obtained with glycosidation of one of the hydroxyl groups
located at positions 5 and 6.
Classification

1. They can be divided into two structural groups, the yellow betaxanthins (from
Latin beta, red beet and Greek xanthos, yellow) and red-purple betacyanins
(kyanos, blue color), depending on R1-N-R2 moieties. More than 50 betalains
are well known, and all of them have the same basic structure, in which R1
and R2 may be hydrogen or an aromatic substituent. Their color is
attributable to the resonating double bonds. Conjugation of a substituted
aromatic nucleus to the 1,7-diazaheptamethinium chromophore shifts the
absorption maximum from 480 nm in yellow betaxanthins to 540 nm in red-
purple betacyanins.
2. In some earlier papers, the terms “betalaines”, “betacyanines”, and
“betaxanthines” were used; the terminal letter “e” was added by Fisher and
Dreiding148 so the names would conform to the I.U.P.A.C. nomenclature; at
present, these terms can be used without the terminal “e”.
3. Betacyanins and betaxanthins can be classified using their chemical
structures. Betacyanin structures show variations in their sugar (e.g., 5- O– D-
Glucose) and acyl groups (e.g., feruloyl), whereas betaxanthins show
conjugation with a wide range of amines (e.g., glutamine) and amino acids
(e.g., tyrosine) in their structures.
Distribution

Among higher plants the occurrence of betalains is restricted to the

Caryophyllalesand those found in certain higher fungi such as Amanita, Hygrocybe,
and Hygrosporus . Betalains of higher plants are in different organs. They produce
red, yellow, pink, and orange colors in Aizoaceae and Portulacaceae flowers, and
purple pigmentation in Cactaceae fruits and in red-beet root (Chenopodiaceae).
Betalains are in bracts, for example, Bougainvillea (Nyctagynaceae) possesses a
wide range of colors; they are also in seeds of Amaranthus, in leaves of Teloxis and
in stems.

Common names and classification of different betacyanins and betaxanthins are

standardized, and they are usually assigned in agreement with their botanical
genus. In the betacyanin group, amaranthin-I was obtained from Amaranthus
tricolor, betanin from Beta vulgaris , and gomphrenin-I from Gomphrena globosa .
While in the betaxanthin group, miraxanthin occurs in flowers of Mirabilis jalapa,
vulgaxanthinI and II have been found in root of Beta vulgaris, and portulaxanthin
has been isolated from the petals of Portulaca grandiflora .

Up to date more than structures of naturally occurring betalains have been

identified. A considerable number of different betacyanins may be derived from two
basic compounds, betanidin (2S, 15S) and isobetanidin (2S, 15R) by glycosidation of
one of the hydroxyl groups located at position 5, for example, betanin, which occurs
as the 5- O-glucoside, and the less-occurring position 6, for example, gomphrenin-II,
which is a 6- O-glucoside. No betacyanin is known to have both positions substituted
with sugar residues. A few biosides are known: amaranthin the betanidin 5- O-[2– O
- (β - D-glucopyranosyluronic acid) β - D-glucopyranoside], and its epimer
isoamaranthin is in the leaves of A. caudatus . Moreover, two epimeric betacyanins,
bougainvillein-r-I and isobougainvillein-r-I, have been isolated from B. glabra .

The isomeric 5- O - β-cellobiosides have been produced by deacylation of the

pigments aleracinI and II from P. oleracea. The only known 6-(2 G-
glucosylrutinosides) of betanidin and isobetanidin have been obtained by
deacylation of bougainvillein-V. Betalain glycosides can be esterified with
hydroxycinnamic acids (ferulic and pcoumaric acids), for example, celosianin-I 4-
coumaroylamaranthin) in Chenopodium rubrum and Lampranthus sociorum ; in
addition, sulfuryl and malonyl betacyanins are known, for example, rivianin
(betanin-3 ′-sulfate) from Riviana humilis and phyllocactin (malonic acid 6 ′-half-
ester of betanin) from Phyllocactus hybridus . The decarboxybetanidin, isolated
from Carpobrotus acinaciformis, is unique among betacyanins in containing a
modified aglycone moiety.

There are about 15 naturally occurring betaxanthins; the indicaxanthin from

Opuntia ficus-indica was the first crystallized. In total, eight of the naturally
occurring betaxanthins contain non-protein amino acids.
Physical Properties

Properties Betalaines absorb light strongly. The absorptivity value(A1% 1cm) is

1120 for betanin and 750 for vulgaxanthine, suggesting high tinctorial strength in
the pure state. The color of betanin solutions is influenced by pH. The spectra of
betanine solutions at pH values between 3.5 and 7.0 do not change, and they
exhibit maximum light absorbance at 537–538 nm. No change in hue occurs
between these pH values. Below pH 3.0, the absorption maximum shifts toward a
slightly shorter wavelength (535 nm at pH 2.0) and the color shifts toward violet.
Above pH 7.0, the absorption maximum shifts toward a longer wavelength (544 nm
at pH 9.0) and the color shifts toward blue. Von Elbe et al. (1974) found that the
color of betanin is most stable between pH 4.0 and 6.0. The thermostability is
greatest between pH 4.0 and 5.0. Light and air have a degrading effect on betanin,
and the effect is cumulative.
Biosynthesis: Biochemistry

1. Biochemistry

Initial stages- The determination of the chemical structure of betalains and of their
biosynthetic intermediates contributed to the establishment the corresponding
biosynthetic pathway; in addition, feeding experiments with isotopically labeled
precursors and in vitro cell cultures were important tools in such discovering.
However, very few enzymes involved in betalain synthesis have been purified and
characterized.

In detail, betalains are considered secondary metabolites; they derive from shikimic
acid and from the tyrosine aminoacid. In their basic structure, the phenyl group is
bonded to a lateral n-propyl chain giving place to a C6-C3 unit. Biogenesis of
betalains from tyrosine has not been thoroughly understood, and only a few
enzymes involved in the biosynthetic route have been identified.
Pathway proposed for betalain biosynthesis. (A) Intial stage, (B) betacyanin biosynthesis. Betacyanins
may be glycosylated at position R1 and R6 and or acyglycosylated at postion R3 or R4. (B) Betaxanthin
biosynthesis. R5 and R6 represent lateral chains of amine compounds. Some of the enzymatic activities
involved in betalain biosynthesis are (1) DOPA 4,5-dioxygenase, (2) glycosyltransferase, and (3)
acyltransferase
Two molecules of tyrosine are required in the biosynthesis of one molecule of
betacyanin or betaxanthin. Initially, two molecules of 3-hydroxyL-phenylalanine (L-
DOPA) are formed. The hydroxylation of tyrosine to L-DOPA is recognized as the first
step in the biogenesis of betalains because experiments with radioactive precursors
have shown good incorporation of labeled [C14]- tyrosine into amaranthin and
betanin, the major betacyanins in Amaranthus tricolor seedlings and red-beet (Beta
vulgaris), respectively. The assumption has been that the first enzyme is a phenol-
oxidase complex catalyzing both the conversion of tyrosine to L-DOPA by a
monophenol oxidase and the dehydrogenation of the latter to O-quinone by a
diphenol oxidase.

Betacyanin biosynthesis- Betacyanins are formed through the reaction of cyclo-

DOPA with betalamic acid followed by glycosylation, or by condensation of the
cycloDOPA glycosides with betalamic acid. In general, it could be mentioned that
condensation of betalamic acid with cyclo-DOPA to betanin formation and the
subsequent glucosylation reaction to betanidin 5-O-β-glucoside, main compound of
red-beet, remain unknown. Complete glycosylation takes place with cyclo-DOPA
followed by condensation with betalamic acid. On the other hand, free betanidin can
be stored as the main component in betalain-producing cells, and it has been shown
that betanidin can be the main receptor of UDP-glucose from glucosyl-transferase
during betanin biosynthesis. It may be possible that the sequence of reactions
depends on the plant genus.

Betanin glucosylation catalyzed by uridine 5′-diphospho-glucose: betanidin 5-O-

βglucosyltransferase (5-GT) was demonstrated by Heuer and Strack,208 who
described the occurrence of one of the two proposed pathways of betacyanin
formation, the transfer of glucose to 5-hydroxy group of betanidin in the formation
of betanin.

Betaxanthin biosynthesis- Little is known about the betaxanthin biosynthesis.

However, it has been suggested the interchange of basic compounds as one of the
main routes. A spontaneous condensation between betalamic acid and an amine
group inside the vacuole has been based on genetic and biochemical studies on
clones of P. grandiflora. In a recent work, Hempel and found two new betaxanthins
in hairy root cultures of Beta vulgaris var. Lutea, vulgaxanthin III, and IV, when the
culture media was supplemented with the corresponding L-amino acids. This
feeding experiment provides arguments for a spontaneous condensation of
betalamic acid with amino acids or amines in the course of betaxanthin biosynthesis

2. Biosynthesis Regulation

In plants, betalain biosynthesis is subject to complex regulation. Pigments are

accumulated only in certain tissues and at specific stages of development. Their
synthesis has been shown to be regulated by light and cytokinins. It has been
shown that effects of DOPA, light, and kinetin exposure are different between
models producing betacyanins or betaxanthins. The betanin synthesis was only
induced under the influence of DOPA or with the combination of kinetin and light.
On the other hand, betaxanthins were synthesized only when the seedlings were
fed with DOPA, and a significant increment of this production was obtained by light
and kinetin supplementation.

The presence of free betalamic acid in plants, which produce betaxanthins or

betacyanins and betaxanthins, and its absence in plants, which produce only
betacyanins, suggest a regulatory mechanism during its biosynthesis. A coordinated
condensation mechanism of betalamic acid with cyclo-DOPA or glycolized-cyclo-
DOPA has been suggested in plants that produce betacyanins; then the
accumulation of betalamic acid is arrested. On the other hand, control mechanisms
in betaxanthin-producing plants must be different to allow the accumulation of
betalamic acid. Regulatory mechanisms on betalain formation are largely unknown,
but the involvement of photoregulation and hormone control has been suggested.

Cell lines produced from red beet showed a range of cell colors via specific
induction methods. Phenotypic color ranged from white/green through yellow,
orange and red to deep violet, representing all types of pigments found in redbeet
plant. Differences in callus phenotypes were associated with cells of markedly
different morphologies; cells were classified into two groups: white, orange, and
violet phenotypes, and green yellow and red phenotypes. The selective expression
of betaxanthin and betacyanin appeared to occur through a limited number of
discrete, stable, and differentiated states, because only four colored phenotypes
were isolated.

In absence of specific information, it is only possible to speculate about the

regulatory mechanism affecting the gene expression during phenotypic transitions
of plant cell cultures. It has been proposed that DNA transposition or specific
rearrangements such as translocations, inversions, breakage, and fusion are
involved. It has been also suggested that such rearrangements could result in the
relocation of particular genes, either within the same chromosome or to another
chromosome. Moreover, it is possible that regulation of pigment synthesis is so
tightly coupled to cellular morphology that an enhancement of gene expression
never occurs in the yellow and red cells.
Functions

a. Taxonomic Markers

Even before the structure of betalains was evident, the importance of betalain
pigments in plant taxonomy and systematic distribution was clear. Betalains are in
eight families: Amaranthaceae, Aizoaceae, Basellaceae, Chenopodiaceae,
Cactaceae, Nyctaginaceae, Phytolaccaceae, and Portulacaceae. Nowadays, it is
known that 9 of 11 families of the Caryophyllales order contain betalains. The
recent addition to the list of betalain families is Didieraceae, a small family from
Madagascar.

Only two families of Caryophyllales, viz., Caryophyllaceae and Molluginaceae, lack

betalains and possess anthocyanins. This suggests an early differentiation of the
Caryophyllales into groups with different kinds of pigments.
RosendalJensen et al. constructed a phenogram that shows the relationship
between betalains, glucosinolates, and polyacetylenes in flowering plants. This
remarkable correlation between chemical and morphological characters has led to
propose that the order Centrospermae, including Cactaceae, must be reserved for
betalain-containing families, while the anthocyanin-containing ones
(Caryophyllaceae and Molluginaceae) must be separated into the related but
distinct order Caryophyllales.

b. Ecological and Physiological

Aspects As in the case of other secondary metabolites, it is impossible to assign a
definite function to betalains in the economy of the organisms that produce them.
When pigments are in flowers or fruits they may have a role as attractants for
vectors (insects or birds) in the pollination process and in seed dispersal by animals,
such as anthocyanins.
The occurrence in other plant parts (e.g., leaves, stem, root) may be devoid of
immediate function. However, it has been suggested that betalain accumulation in
red beet root is related to the storage of carbohydrates as a physiological response
under stress conditions. In addition, the transient coloration of many seedlings and
the reddening of senescent leaves of several plants of Caryophyllales order (e.g.,
Kochia scoparia) have no obvious physiological or ecological reasons. Whatever its
significance, the process the analogous phenomenon observed in anthocyanin-
producing species. Betalains are also produced in injured tissues, normally not
pigmented, possibly as a defense mechanism against infection. This physiological
response was only observed in plants possessing specific factors that have been
associated with two novel antifugal proteins.
Interestingly, it must be mentioned that betalains from Beta vulgaris (betanin and
vulgaxanthin) are effective inhibitors of indoleacetic acid (IAA) oxidase and that
betanin counteracts the inhibitory effect of IAA on wheat root elongation. It means
that betalains could be considered as potential modifiers of auxin metabolism;
notwithstanding, there is no evidence that supports the betalains role as in vivo
regulators of IAA activity

c. Pharmacological Effects
Although structurally related to alkaloids, betalains have no toxic effects in the
human body, as can be deduced from the fact than they are present in considerably
high amounts in certain foodstuffs, such as red-beet, prickly pear fruits, and
Amaranthus seeds. Therefore, betalains represent a safe natural alternative to
some synthetic color additives that are currently in use. Interestingly, there is no
upper limit to the recommended daily intake.
Betanin has shown antiviral and antimicrobial activities (e.g., Pythium debaryum, a
pathogenic fungi in red-beet). In some places in Mexico, an infusion of Bougainvillea
bracts mixed with honey is used widely for a cough.
Finally, in a recent work, the importance of some natural pigments as nutraceutical
ingredients was reviewed. It was suggested that betalains like anthocyanins, β-
carotene, and various vegetable and fruit extracts must be used for their potential
health benefits. For example, yellow betaxanthins, in addition to their potential role
as natural food colorant, may be used as a means of introducing essential dietary
amino acids into foodstuffs, giving rise to an “essential dietary colorant”. Another
new interesting area includes foods that can generate their own light (“biolume
products”) in which betalains could play an important role, suggesting how exciting
the area of natural colorants has become.
Methodological Aspects

a. Extraction

Betalains-containing material (raw plant or cell culture) are generally macerated or

ground. Pigments can be extracted with pure water, cold, or at room temperature,
although in most cases the use of methanol or ethanol solutions (20 to 50% v/v) is
necessary to achieve complete extraction. Sometimes, the necessity of an aerobic
juice fermentation (e.g., Saccharomyces cerevisiae, Aspergillus niger) in order to
reduce free sugars and then to increase the betacyanin content has been reported.
In both procedures, the inactivation of degradative enzymes by a short heat
treatment of the extract (70ºC, 2 min) could be desirable, although this may destroy
some of the pigments. Betacyanins can be precipitated by a slight acidification with
hydrochloric acid or with acidified ethanol (0.4 to 1% HCl); subsequently, by the
addition of 95% aqueous ethanol yields betaxanthins.
Degradation of betanin may occur very fast and destruction of complex pigments
should also be avoided, because acid-acylated betacyanins are rapidly deacylated
and such pigments can be over looked. In such a situation, extraction should be
carried out with cold water for long-term and darkness conditions.

b. Separation
 Ion-exchange and column chromatography.
Ionic-exchangers are the most widely used adsorbents in fractionation, as much
as in separation; then gel filtration is used. In a simple and rapid procedure,
plant extract must be stirred with the ion-exchanger resin (e.g., Dowex 50W-X2,
Merck I, DEAE-Sephadex A25, etc.), which adsorbs the betalains (nonionic
interaction). Subsequently, resin is washed with aqueous HCl (0.1% v/v) and
pigments are eluted with water followed by final separation on a
chromatographic column (e.g. Polyamide, Polyclarc-AT, or polyvinylpyrrolidone,
Sephadex G-15 and G-25).
 Electrophoresis and thin layer chromatography (TLC).
Paper electrophoresis using pyridine and formic or acetic acid as solvents or in
cellulose are common and reliable methods for betacyanin detection, because
they migrate first as immobile zwitterions (pH 2), followed as monoanions (pH 2
to 3.5), and finally as bisanions (pH 3.5, 7.0). In the case of betaxanthins, the
mobility may be related to indicaxanthin, and betacyanins are related with the
mobility of betanin. Electrophoresis can be carried out using pyridine-citric acid
solvent, voltage gradient of 5.6 volts/cm, and a temperature of 4ºC.
 High-performance liquid chromatography (HPLC).
The HPLC technique has become the method of choice for chromatographic
separation, rapid quantification, and tentative identification of betalains.
 Chemical properties

a. Stability
When betalains are used as food colorants, color stability is a major concern. There
are several factors that have been recognized to affect the stability of these
pigments:
 pH. The hue of betalains is unaffected at the pH between 3.5 to 7; the values
of most foods are in this range. Betalain solutions in this pH range showed a
similar visible for betacyanins and betaxanthins. Betacyanin maximum is in
the wavelength (λ) range 537 to 538 nm, while betaxanthin maximum is
between 475 to 477.497 Below pH 3.5, λ shifts toward a lower wavelength,
and above 7 the change is toward a longer wavelength; out of the pH range
3.5 to 7 the intensity of the visible spectra decreases. Stability of betanin
solutions is pH dependent. Optimal pH for maximum betanin stability in the
presence of oxygen is between 5.5 to 5.8. Red beet solutions showed their
maximum stability at pH 5.5, the normal pH for beets.
 Temperature. Temperature also shows a clear effect on betalain stability.
Thermal kinetic degradation of betanin has been evaluated by several
authors. It has been reported that thermostability of betanin solutions is pH
dependent and partially reversible. Heating of betanin solutions produces a
gradual reduction of red color, and eventually the appearance of a light
brown color.
 Light. Von Elbe et al.495 found that rate of betanin degradation increased
15.6% after pigment daylight exposure at 15ºC. Degradation of light-exposed
betalains followed a first-order kinetic. In addition, it was observed that
degradation was higher at pH 3.0 (k = 0.35 days-1) than at pH 5.0 (k = 0.11
days-1), when betacyanins were exposed to fluorescent light. On the other
hand, at darkness conditions betacyanins were most stable ( k = 0.07 days^-
1). Attoe and von Elbe showed an inverse relationship between betalain
stability and light intensity in the range 2200 to 4400 lux). It is explained that
visible light absorption excites π electrons of the pigment chromophore to a
more energetic state (π*). This would cause a higher reactivity or a lowered
activation energy for the molecule (E A = 25 Kcal·mol^-1 in darkness and
19.2 in illumination)
 Water activity. The degradation reaction involves water, the greatest
stability of betalains has been reported in foods or model systems of low
moisture and a w .Pigment degradation follows first-order kinetics, and
stability increases with decreasing aw. It has been established that a w has a
pronounced exponential effect on pigment stability. Pigment stability
decreases in one order of magnitude when aw was increased from 0.32 to
0.75.
 Oxygen . Oxygen causes a product darkening and loss of color. Betanin
reacts with molecular oxygen, producing pigment degradation in air-
saturated solutions. Degradation kinetic under air atmosphere follows a first-
order model, but deviates from first-order in the absence of oxygen. Betanin
degradation is a partially reversible reaction, and it has been reported that in
order to increase the recovering of pigment it is necessary to have the
samples under low levels of oxygen. Thus, heated betanin solutions (pH 4.75,
130 min, 15ºC) under low oxygen levels showed an increased betanin
retention from 54 to 92%.
 Several methods have been reported to prevent the destruction or to
improve the stability of pigments, including degassing, addition of
antioxidants and stabilizers, control of pH, minimal heath treatment, among
others, and these efforts have been directed to their application in food
products.

b. Stability during Processing operations and Stability in Foods

The sensitivity of betalains to different factors suggests that their application as

food colorants is limited. Based on these properties, betalains can be used in
foods with a short shelf-life, produced by a minimum heat treatment, and
packaged and marketed in a dry state under reduced levels of light, oxygen, and
humidity.
Betalains have several applications in foods, such as gelatins desserts,
confectioneries, dry mixes, poultry, dairy, and meat products.
The amount of pure pigment required in these foods groups to obtain the desired
hue is relatively small and for most applications does not exceed 50 ppm of
betalains, calculated as betanin. Problems associated with betalain degradation
and pigment recovery during the processing operations are of economic
importance and must be solved to betalains displace the application of synthetic
dyes in some food products. The effectiveness of commercial betalains depends
largely on a continuous availability of highly pigmented sources, the use of cold
and modified storage atmospheres prior to processing, efficient enzymatic
control, handling practices, extraction procedures, purification, concentration,
and finishing operations e.g., freeze, spray, and vacuum drying).
Nowadays, beet roots represent the main commercial source of betalains
(concentrates or powders). Many factors during the pre- and postharvest period
and during processing influence the recovery of these natural beet colorants. In
addition, recent efforts are centered around the betalain content in red beets
through selective breeding. Initially, high pigment content is very important. The
average pigment content of beets is approximately 130 mg/100 g fresh weight,
but new red beet varieties produce around 450 to 500 mg/100 g fresh weight.
Furthermore, this value is increasing as advanced selection is developed.
Commercial preparations of beet pigment for use as food colorants are available
as either juice concentrates (produced by concentrating juice under vacuum to
60 to 65% total solids) or powders (produced by freeze or spray drying). These
preparations contain from 0.3 to 1% of pigment. They show a variety of colors,
depending on their content of yellow pigments, and may have a beet-like odor
and flavor. The remainder of the solids is mainly sugars (75 to 80%), ash (8 to
10%), and protein (10%). On a laboratory scale, betalains can be obtained by
employing reverse osmosis, ultrafiltration, solid -liquid extraction, and dif fusion.
These processes have been shown to be efficient on the recovery of betalains
from raw beet tissue when compared with conventional hydraulic techniques. As
approximately 80% of beet juice solids consist of fermentable carbohydrates and
nitrogenous compounds, a fermentation process to remove these materials has
been widely employed. The yeast Candida utilis and Saccharomyces cerevisiae
have been used in the fermentative process, whereas a strain of Aspergillus
niger not only destroyed free sugars but also enriched the colorant in betanin.
The powder obtained from fermented juice contained five to seven times as
 much as the betacyanin obtained in the powder from raw juice (on a dry weight
basis).

c. Production of Betalains by Plant Tissue Culture

Cell tissue culture has been a very useful tool in the study of various aspects of
biochemistry, enzymology, genetics, and biosynthesis of betalains, and,
interestingly, betalain production by plant cell culture will represent an excellent
option in the future; it has a number of advantages over conventional
procedures. Mainly with this methodology, it is possible to control quality and
availability of pigments independently of environmental changes

BIBLIOGRAPHY
1. Food Chemistry: Third Edition Edited by Owen R. Fennema
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Chang Yong Lee
3. Natural Pigments: Carotenoids, Anthocyanins, and Betalains — Characteristics, Biosynthesis,
Processing, and Stability Review article by F. Delgado-Vargas, A. R. Jiménez,and O. Paredes-
López