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SPIRULINA PLATENSIS – FOOD FOR FUTURE: A REVIEW

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Abstract

Spirulina can play an important role in human and animal nutrition, environmental protection through wastewater recycling and energy conservation. The present review was focused on the various characteristics of Spirulina platensis. Spirulina is rich in proteins (60-70%), vitamins and minerals used as protein supplement in diets of undernourished poor children in developing countries. One gram of Spirulina protein is equivalent to one kilogram of assorted vegetables. The amino acid composition of Spirulina protein ranks among the best in the plant world, more than that of soya bean. The mass cultivation of Spirulina is achieved both in fresh water and waste water. Spirulina grown in clean waters and under strictly controlled conditions could be used for human nutrition. The micro alga grown in waste water is used as animal feed and provide a source of the fine chemicals and fuels. The waste water system is highly applicable in populated countries like India where wastes are generated in high quantities and pose environmental problem. The present review focused the following topics: Spirulina platensis, Isolation and occurrence of Spirulina platensis, newly formulated media for Spirulina cultivation, Phycocyanin and Medicinal properties of Spirulina platensis.
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Asian Journal of Pharmaceutical Science & Technology
e-ISSN: 2248 9185
www.ajpst.com Print ISSN: 2248 9177
SPIRULINA PLATENSIS FOOD FOR FUTURE: A REVIEW
P. Saranraj* and S. Sivasakthi
Department of Microbiology, Annamalai University, Annamalai Nagar, Chidambaram-608 002, Tamil Nadu, India.
ABSTRACT
Spirulina can play an important role in human and animal nutrition, environmental protection through wastewater
recycling and energy conservation. The present review was focused on the various characteristics of Spirulina platensis. Spirulina
is rich in proteins (60-70%), vitamins and minerals used as protein supplement in diets of undernourished poor children in
developing countries. One gram of Spirulina protein is equivalent to one kilogram of assorted vegetables. The amino acid
composition of Spirulina protein ranks among the best in the plant world, more than that of soya bean. The mass cultivation of
Spirulina is achieved both in fresh water and waste water. Spirulina grown in clean waters and under strictly controlled
conditions could be used for human nutrition. The micro alga grown in waste water is used as animal feed and provide a source of
the fine chemicals and fuels. The waste water system is highly applicable in populated countries like India where wastes are
generated in high quantities and pose environmental problem. The present review focused the following topics: Spirulina
platensis, Isolation and occurrence of Spirulina platensis, newly formulated media for Spirulina cultivation, Phycocyanin and
Medicinal properties of Spirulina platensis.
Key words: Spirulina platensis, Single cell protein, Protein content and Phycocyanin.
INTRODUCTION
Blue-green algae (cyanobacteria) are among the
most primitive life forms on Earth. Their cellular structure
is a simple prokaryote. They share features with plants, as
they have the ability to perform photosynthesis. They share
features with primitive bacteria because they lack a plant
cell wall. Interestingly, they also share characteristics of the
animal kingdom as they contain on their cellular membrane
complex sugars similar to glycogen. Among blue-green
algae, both edible and toxic species adapted to almost any of
the most extreme habitats on earth. Edible blue-green algae,
including Nostoc, Spirulina, and Aphanizomenon species
have been used for food for thousands of years [1].
The current environmental conditions deteriorations, mental
and physical stress, changes in the diet have been serious
risk factors for the humans, increased the death rate and
civilization diseases. These are the obvious reasons why
new progressive trends are being extensively developed in
modern medicine, pharmacology and biotechnology and
more effective harmless medicaments are being sought for
to treat and prevent various diseases. One of the trends in
biotechnology is associated with Blue green microalgae
Spirulina platensis which have been widely employed as
food and feed additives in agriculture, food industry,
pharmaceuticals, perfume making, medicine and science
[1].
Spirulina platensis
Spirulina sp. has been used as food for centuries by
different populations and only rediscovered in recent years.
Once classified as the ―blue-green algae’’, it does not
strictly speaking belong to the algae, even though for
convenience it continues to be referred to in that way. It
grows naturally in the alkaline waters of lakes in warm
regions. Measuring about 0.1mm across, it generally takes
the form of tiny green filaments coiled in spirals of varying
tightness and number, depending on the strain. Its
impressive protein content and its rapid growth in entirely
mineral environments have attracted the attention of both
researchers and industrialists alike.
Spirulina are unicellular and filamentous blue-
green algae that has gained considerable popularity in the
health food industry and increasingly as a protein and
vitamin supplement to aquaculture diets. It has long been
used as a dietary supplement by people living close to the
alkaline lakes where it is naturally found. Spirulina has been
used as a complementary dietary ingredient of feed for fish,
shrimp and poultry. Among the various species of Spirulina,
the blue green alga Spirulina platensis has drawn more
attention because it shows an high nutritional content
Corresponding Author: P. Saranraj E-mail: microsaranraj@gmail.com
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characterized by a 70% protein content and by the presence
of minerals, vitamins, amino acids, essential fatty acids etc
[2]. P. J. Turpin in the year 1827 isolated Spirulina
from a fresh water sample. In 1884, Wittrock and Nordstedt
reported the presence near the city of Montevideo of a
helical, septal and green-blue microalgae called Spirulina
platensis. In 1852, Latter Stizenberger gave this new genes
the name Arthrospira based on the septa presence, helical
form and multicellular structure. Because of the common
helical morphology, reunified the members of the two
genera under the designation Spirulina without considering
the septum, only morphological similarity. In 1989, these
microorganisms were separately classified into two genera
Spirulina and Arthrospira; this classification is currently
accepted.
According to the classification in Bergey’s Manual
of Determinative Bacteriology, Spirulina belongs to the
oxygenic photosynthetic bacteria that cover the groups
Cyanobacteria and Prochlorales, which are, by phylogeny,
related to the sequence of the rRNA (ribosomal ribonucleic
acid) sub-unit 16s. As a function of the sequence data of this
sub-unit and the rRNA sub-unit 5s, these prokaryotes are
classified within the Eubacteria group.
The dried cells of microorganisms such as bacteria,
fungi, yeasts and algae that are grown in large scale culture
systems as proteins, for human or animal consumption are
collectively known as single cell protein. SCPs are
characterized by; fast growth rate; high protein content (43-
85%) compared to field crops; require less water and land
and independent of climate; grow on wastewater; can be
genetically modified for desirable characters such as amino
acid composition and temperature tolerance. Among the
various microorganisms used as sources of SCP, the blue
green algae, Spirulina is considered as the best source. The
composition of the biomass, including the high protein
content, low content in nucleic acids, occurrence of high
concentrations of vitamins and other growth factors and the
presence of cell wall that is more easily digestible than that
of other microbes indicate that Spirulina is a promising
source of food or feed.
Spirulina platensis is naturally found in tropical
regions inhabiting alkaline lakes (pH 11) with high
concentration of NaCl and bicarbonates. These limiting
conditions for other microorganism allow cultivation of
microalgae in opened reactors [3]. In Cyanobacteria, the
light harvesting pigments include chlorlphyll-a, carotenoids
and phycobiliproteins. The later are proteins with linear
tetrapyrrole prosthetic groups called according to their
structure: phycocyanin, phycoerythrin and allophycocyanin
[4].
3. ISOLATION AND OCCURENCE OF Spirulina
platensis
Spirulina platensis is naturally found in tropical
regions inhabiting alkaline lakes (pH 11) with high
concentration of NaCl and bicarbonates. These limiting
conditions for other microorganism allow cultivation of
microalgae in opened reactors [5]. In Cyanobacteria, the
light harvesting pigments include chlorophyll-a, carotenoids
and phycobiliproteins. The later are proteins with linear
tetrapyrrole prosthetic groups called according to their
structure: phycocyanin, phycoerythrin and allophycocyanin
[6]. Spirulina is commonly found in aquatic
ecosystems like lakes, ponds and tanks. It is one of the
nature’s first photosynthetic organisms capable of
converting light directly for complex metabolic processes.
Spirulina is used for food from time immemorial by tribes
living around Chad Lake in Africa. The predominant
species of phytoplankton of the lake is Spirulina platensis.
The algae Spirulina was eaten in Mexico under the names
‘Tecuitlatl’ [7]. Spirulina grows optimally in pH range of 9-
11 and there is least chance of contamination of other
microbes.
Orio Ciferri [8] studied that Spirulina was a
ubiquitous organism. After the first isolation by Turpin in
1827 from a fresh water stream, species of Spirulina have
been found in a variety of environments: soil, sand,
marshes, brakish water, sea water, and fresh water. Species
of Spirulina have been isolated, for instance, from tropical
waters to the North sea, thermal springs, salt pans, warm
waters from power plants, fish ponds, etc. Thus, the
organism appears to be capable of adaptation to very
different habitats and colonizes certain environments in
which life for other microorganisms is, if not possible, very
difficult; typical is the population by alkalophylic Spirulina
platensis of certain alkaline lakes in Africa and by Spirulina
maxima of lake Texcoco in Mexico. In some of these lakes
Spirulina grows as a quasimonoculture.
Susan Spiller et al. [9] examined the fine structure
of Cyanobacteria, Spirulina platensis and Spirulina subsala
as viewed by X-ray microscope, XM-1, beamline 6.1.2. in
the initial stages of a project to obtain high resolution
images of these spiral Cyanobacteria and the spiral
chloroplasts of Spirogyra species. These high resolution
images will allow a close comparison between
morphological features of prokaryotic cells and the
eukaryotic photosynthetic organelle. Cryo-technique are
being developed with the cryo-stage XM-1, the X-ray
microscope at beam line 6.1.2 of the advanced tight source.
The bright blue-green cylindrical filaments of Spirulina
platensis had a lazy spiral turn, filament width 5-6 µm. Cell
walls are clearly visible crossing the filament at intervals of
about 2-3µm.
Martha et al. [10] studied that before Columbus,
Mexicans (Aztecs) exploited this microorganism as human
food; presently, African tribes (Kanembu) use it for the
same purpose. Its chemical composition includes proteins
(55%-70%), carbohydrates(15%-25%), essential fatty
acids(18%) vitamins, minerals and pigment like chlorophyll
a and phycocyanin. The last one is used in food and
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cosmetic industries. Spirulina is considered as an excellent
food, lacking toxicity and having corrective properties
against viral attacks, anemia, tumor growth and
malnutrition. It has been reported in literature that the use of
these microalgae as animal food supplement implies
enhancement of the yellow coloration of skin and eggs yolk
in poultry and flamingos, growth acceleration, sexual
maturation and increase of fertility in cattle.
NEWLY FORMULATED MEDIA FOR SPIRULINA
CULTIVATION
Basirath Raoof et al. [11] formulated a new
medium for mass production of Spirulina sp. by
incorporating selected nutrients of the standard Zarrouk’s
medium. This newly formulated medium contains single
super phosphate, sodium nitrate, muriate of potash, sodium
chloride, magnesium sulphate, calcium chloride and sodium
bicarbonate (commercial grade). Maximum growth rate in
terms of dry biomass, chlorophyll and proteins in SM was
recorded between 6 and 9 days 0f growth and values were
0.114, 0,003, and 0.068 as compared to 0.112, 0.003 and
0,069 mg/ml significant differences were observed in the
protein profiles of Spirulina sp. grown in both the media.
From the scale up point of view, the revised medium was
found to be highly economical, since it is five times cheaper
than Zarrouk’s medium.
Dao-Lun et al. [12] attempted to culture Spirulina
platensis in human urine directly to achieve biomass
production and O2 evolution, for potential application to
nutrient regeneration and air revitalization in life support
system. The culture results have showed that Spirulina
platensis in diluted human urine was lighter than that in
Zarrouk’s medium.
Raoof et al. [13] investigated the use of a new
medium formulated for mass production of Spirulina sp. by
incorporating selected nutrients of the standard Zarrouk’s
medium (SM) and other cost-effective alternative
chemicals. This newly formulated medium (RM6) contains
single super phosphate (1.25 g/litre), sodium nitrate (2.50
g/litre), muriate of potash (0.98 g/litre), sodium chloride
(0.50 g/litre), magnesium sulfate (0.15 g/litre), calcium
chloride (0.04 g/litre), and sodium bicarbonate (commercial
grade) 8 g/litre. The alga was grown in an illuminated (50
mmol photons/m²/s white light) growth room at 30 ± 1 °C.
Maximum growth rate in terms of dry biomass, chlorophyll
and proteins in SM was recorded between 6 and 9 days of
growth and values were 0.114, 0.003 and 0.068 as compared
to 0.112, 0.003 and 0.069 mg/ml/day in RM6. No
significant differences were observed in the protein profiles
of Spirulina sp. grown in both the media.
Harriet Volkmann et al. [14] cultivated the
Spirulina platensis in laboratory under controlled conditions
(30ºC, photoperiod of 12 hours light/dark provided by
fluorescent lamps at a light intensity of 140 μ mol
photons.m-2.s-1 and constant bubbling air) in three different
culture media: (1) Paoletti medium (control), (2) Paoletti
supplemented with 1 g L-1 NaCl (salinated water) and (3)
Paoletti medium prepared with desalinator wastewater. The
effects of these treatments on growth, protein content and
amino acid profile were measured. Maximum cell
concentrations observed in Paoletti medium, Paoletti
supplemented with salinated water or with desalinator
wastewater were 2.587, 3.545 and 4.954 g L-1, respectively.
Biomass in medium 3 presented the highest protein content
(56.17%), while biomass in medium 2 presented 48.59%
protein. All essential amino acids, except lysine and
tryptophan, were found in concentrations higher than those
required by FAO.
Bohra [15] investigated growth pattern of Spirulina
platensis in standard and modified media based on
seawater-chemicals and seawater fertilizers. During the
cultivation, the cell concentrations were analyzed at 540nm
along with protein and chlorophyll a estimation. Growth
patterns of different species and strains were monitored for
25 days and specific growth rate, mean daily division rate
and doubling time were calculated. Spirulina platensis was
observed to have different specific growth characteristics in
different media at same environmental parameters
(Temperature, pH and light intensity). Even though,
Spirulina platensis in standard media exhibited good growth
patterns, biomass, protein content and chlorophyll content
than other seawater based media, the experiment discusses
the feasibility of seawater based media.
Usharani et al. [16] collected the water samples
for the isolation of strains of alga Spirulina platensis from
three different locations and the strain Spirulina platensis
was isolated and it was, designated as ANS - 1 strain. The
characteristics of Spirulina platensis ANS -1 were
compared with reference CAS -10. The waste water rice
mill effluent was collected and its pH was adjusted to 9-11
by using sodium bicarbonate @ 800 mg l-1and it was used as
a medium. The isolated strain ANS -1 and reference strain
CAS 10 were grown in substrates 1/6 diluted Zarrouk’s
medium (control) and rice mill effluent. The well performed
strain CAS 10 under in vitro condition was selected as
efficient one. The growth of Spirulina platensis was
measured both in laboratory and outdoor condition by using
the parameters viz., optical density, population, dry weight,
protein and chlorophyll content. The high growth and dry
weight were recorded in 1/6 diluted Zarrouk’s medium
when compared to rice mill effluent medium. Maximum
protein and chlorophyll content were noticed in 1/6 diluted
Zarrouk’s medium than rice mill effluent [17].
Lignite fly ash (LFA) is the by-product of thermal
power station. In India, Neyveli Lignite Corporation (NLC)
liberates tonnes of LFA as a waste product and the liberated
LFA highly pollutes the soil and water bodies. The Blue
Green Algae Spirulina platensis grows well at alkaline pH
and the pH of the LFA is also alkaline. The LFA also
contains an array of micronutrients and macronutrients
which favours the growth of Spirulina platensis. In order to
minimize the environmental pollution and recycle the LFA
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waste, Saranraj et al. [18] was planned to utilize the LFA at
different concentrations for the laboratory cultivation of
Spirulina platensis. Spirulina platensis was cultivated in
the conical flasks containing Zarrouk’s medium alone [SP]
and Zarrouk’s medium with three different concentrations
(0.5 g/l [SP - 1], 1.0 g/l [SP - 2] and 1.5 g/l [SP - 3]) of LFA
supplementation. Among the four different
supplementations used, SP which contained 1.5 g Lignite
fly ash in one litre of Zarrouk’s medium highly induced the
growth and protein content of Spirulina platensis when
compared to other supplementations. The least growth and
protein content was recorded in SP which contains
Zarrouk’s medium alone without LFA supplementation.
PHYCOCYANIN
Phycocyanin is a water soluble blue pigment that
gives Spirulina its bluish tint. It is found in blue green algae
like Spirulina. Phycocyanin is a powerful water soluble
antioxidant, scientists in Spain showed that an extract of
Spirulina containing phycocyanin is a potent free radical
scavenger and inhibits microsomal lipid peroxidation [19].
Phycocyanin in Spirulina that is though to help protect
against renal failure caused by certain drug therapies.
Phycocyanin has also shown promise in treating cancer in
animals and stimulating the immune system [20]. A human
clinical study showed that a hot water extract of Spirulina
rich in phycocyanin increased interferon production and NK
cytotoxicity (cancer killing
Physical and chemical properties of Phycocyanin
Phycobiliproteins are a small group of highly
conserved chromoproteins that constitute the
phycobilisome, a macromolecular protein complex whose
main function is to serve as a light harvesting complex for
the photosynthetic apparatus of cyanobacteria and
eukaryotic groups. The most common classes of
phycobiliproteins are allophycocyanin, phycocyanin and
phycoerythrin all of which are formed by a and b protein
subunits and carry different isomeric linear tetrapyrrole
prosthetic groups (bilin chromophore) which differ in the
arrangement of their double bonds.
Phycocyanin is composed of two dissimilar a and b
protein subunits of 17 000 and 19 500 Da, respectively, with
one bilin chromophore attached to the a subunit (a 84) and
two to the b subunit (b 84, b 155). Pc exists as a complex
interacting mixture of trimer, hexamer and decamer
aggregates. When the secondary, tertiary and quaternary
structures of the protein are denatured, the visible
absorption band as well as the fluorescence will drop in
intensity. The chemical structure of the bilin chromophores
in Pc is very similar to bilirubin, a heme degradative
product. Bilirubin is considered to be a physiologically
important antioxidant against reactive species [22].
Biochemistry of Phycocyanin
The phycocyanins, biliproteins involved in the
light-harvesting reactions, have been resolved by gel
electrophoresis in Spirulina platensis and Spirulina maxima
and isolated from the former. Both c-phycocyanin and
allophycocyanin appear to be oligomeric complexes
composed of at least two different subunits that may be
resolved by electrophoresis under denaturing conditions.
The α- and β-subunits of c-phycocyanin showed mobilities
corresponding to molecular weights of 20,500 and 23,500,
respectively, resulting in an oligomer with a minimum
molecular weight of ca. 44,000. Allophycocyanin was found
to be composed of subunits with molecular weights of ca.
18,000 and 20,000 to give an oligomer with a minimum
molecular weight of ca. 38,000. Absorption and
fluorescence spectra were similar to those reported for c-
phycocyanins and allophycocyanins isolated from other
cyanobacteria. A study of the denaturation and renaturation
of c-phycocyanin indicated the possibility that more than
one chromophore exists in this biliprotein [23].
Extraction and purification of Phycocyanin from
Spirulina platensis
Phycocyanin is a water soluble blue pigment that
gives Spirulina its bluish tint. It is found in blue green algae
like Spirulina. Phycocyanin is a powerful water soluble
antioxidant, scientists in Spain showed that an extract of
Spirulina containing phycocyanin is a potent free radical
scavenger and inhibits microsomal lipid peroxidation [24].
Phycocyanin in Spirulina that is though to help protect
against renal failure caused by certain drug therapies.
Phycocyanin has also shown promise in treating cancer in
animals and stimulating the immune system [25].
Phycocyanin is the bluish pigment used by blue-
green algae to photosynthesize. It accounts for as much as
20% of the protein in cyanobacteria, and attaches itself to
photosynthesizing membranes. C-phycocyanin is
fluorescent, with an extremely high absorbtivity, high
quantum efficiency, a large strokes shift and excitation and
emission bands at visible wavelengths. It is a stable protein
which can be linked to antibody and other proteins by
conventional protein cross-linking techniques without
altering its spectral characteristics.
In Japan, phycocyanin from Spirulina is used as a
natural blue-pigment for food coloring. In addition, owing
to its fluorescence property, pure phycocyanin is used as
labeling substance in immunoassays, microscopy and
cytometry. Moreover, it has been demonstrated that
Spirulina has therapeutic effects against hyperlipidemia.
Therefore, there is a potential for future pharmaceutical use
of Spirulina to produce many health related products. It may
be possible to further enhance the production of certain
compounds such as p-carotene in Spirulina through genetic
engineering. A human clinical study showed that a hot
water extract of Spirulina rich in phycocyanin increased
interferon production and Nk cytotoxicity (cancer killing
cells) when taken orally [26].
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Lorenz [27] reported that Spirulina and other blue-
green algae contain c-phycocyanin, which acts as an
accessory pigment when light energy is captured and
transferred to chlorophyll a. This is a spectrophotometry
method adapted to extract and quantify a relatively pure c-
phycocyanin fraction from Spirulina Pacifica.
Cheyla Romay and Ricardo Gonzalez [28]
experimented the antioxidative property of the phycocyanin.
They used the human erythrocytes for examine the
antioxidant property of phycocyanin. Their results provided
protective effect of phycocyanin against hemolysis induced
by peroxyl radicals in human erythrocytes, which seems to
be due to the scavenging action of the radicals in the
aqueous phase. Lipid peroxidation is inhibited similar to
trolox and ascorbic acid.
Noam Adir et al. [29] studied the crystal structure
of the light-harvesting phycobiliprotein, c-phycocyanin
from the thermophilic cyanobacterium Synechochoccus
vulcanus has been determined by molecular replacement to
2.5 resolution. The crystal belongs to space group R32
with cell parameters a = b = 188.43 A°, c =61.28 A°, α= b
=90°, γ= 120 °, with one (ab) monomer in the asymmetric
unit. The structure has been reined to a crystallographic R
factor of 20.2% (R-free factor is 24.4 %), for all data to 2.5
A°. The crystals were grown from phycocyanin (αβ)3
trimers that form (αβ)6 hexamers in the crystals, in a
fashion similar to other phycocyanins.
Minkova et al. [30] purified C-phycocyanin from
Spirulina (Arthrospira) fusisormis by a multi-step treatment
of the crude extract with rivanol in a ratio (v/v), followed by
40% saturation with ammonium sulphate. After removal of
rivonal by gel filtration on Sephadex G-25, the pigment
solution was saturated to 70% with ammonium sulphate.
After the last step of purification, C-phycocyanin had an
emission and absorption maxima at 620 and 650 nm,
respectively and absorbance ratio A620/A280 of 4.3, which
are specific for the pure biliprotein. Its homogenecity was
demonstrated by sodium dodecyl sulphate-polyacrylamide
gel electrophoresis, yielding two bands of molecular masses
19 500 and 21 500 kDa, corresponding to a and b subunits
of the pigment, respectively. The yield of C-phycocyanin
was: 46% from its content in the crude extract.
Jose et al. [31] extracted astaxantine and
phycocyanin from microalgae with supercritical extraction
and using carbondioxide. The samples of microalgae were
crushed by cutting mills and the manually ground with dry
ice (solid carbondioxide). The Haematococcus extracts were
analysed by liquid chromatography, using astaxantin (purity
of 98%) as a standard. Phycocyanin being insoluble in
carbondioxide was indirectly separated. Thus, lipid-soluble
substances from Spirulina were extracted and analysed by
liquid chromatography. The maximum total recovery of
astaxantin, calculated from its initial and residual content in
the alga (0.0147 and 0.0004, respectively) exceeds 97%, For
phycocyanin extraction, the addition of cosolvent (10 mass
% of ethanol) has a strong effect on the extraction yield of
lipidic substance. The total extraction yield of about 3
mass%. Noam Adir et al. [32] The Crystal Structure of a
Novel Unmethylated Form of C-phycocyanin, a Possible
Connector Between Cores and Rods in Phycobilisomes A
novel fraction of c-phycocyanin from the thermophilic
cyanobacterium Thermosynechcoccus vulcanus, with an
absorption maxima blue-shifted to 612 nm (PC612), has
been purified from allophycocyanin and crystallized. The
crystals belong to the P63 space group with cell dimensions
of with a single monomer in the asymmetric unit, resulting
in a solvent content of 65 %, and diffract to 2.7 . The
PC612 crystal structure has been determined by molecular
replacement and refined to a crystallographic R-factor of
20.9% (R free - 27.8%).
Zhang et al. [33] C-phycocyanin and
allophycocyanin were separated and purified from Spirulina
platensis by precipitation with ammonium sulphate, Ion
exchange chromatography and gel filtration
chromatography. Pure C-phycocyanin and allophycocyanin
were finally obtained with an A260/A280 value of 5.06 and
an A655/A280 value of 5.34, respectively.
Ying Zhang et al. [34] studied the spectral
properties of the glutaraldehyde-treated phycobilisomes.
The results showed that glutaraldehyde was effective in
preventing phycobilisomes from dilution induced
dissociation and preserving the intra-phycobilisomes energy
transfer. Badrish et al. [35] reported that the phycocyanin is
a major phycobiliprotein produced by cyanobacteria, but
only few strains for its efficient purification have been
reported. The phycocyanin was extracted by repeated
freeze-thaw cycles and purity by a three-step process:
ammonium sulphate fractionation, sephadex G-150 size
exclusion chromatography and DEAE cellulose ion
exchange chromatography.
Silveira et al. [36] experimented c-Phycocyanin
extraction from cyanobacteria Spirulina platensis was
optimized using factorial design and response surface
techniques. The effects of temperature and biomass-solvent
ratio on phycocyanin concentration and extract purity were
evaluated to determine the optimum conditions for
phycocyanin extractions. The optimum conditions for the
extraction of phycocyanin from Spirulina platensis were the
highest biomass-solvent ratio, 0.08g/mL/1, and 250C. Under
these conditions it’s possible to obtain an extract of
phycocyanin with a concentration of 3.68mg.mL/1 and
purity ratio (A615, A280) of 0.46.
Silvana et al. [37] presented the evaluation of some
important parameters for the purification of phycocyanin
using ion exchange chromatography. The influences of pH
and temperature on the equilibrium partition coefficient
were investigated to establish the best conditions for
phycocyanin adsorption. The separation of phycocyanin
using the Q-Sepharose ion exchange resin was evaluated in
terms of the pH and elution volume. The highest partition
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coefficients were obtained in the pH range from 7.5 to 8.0 at
250C. Narayan and Raghavarao [38] experiments
aqueous two phase extraction was employed directly to the
cell homogenate of Spirulina platensis for the downstream
processing of C-phycocyanin. This enables integrating the
process steps of cell removal, extraction and concentration
into a single unit operation of aqueous two phase extraction.
The effect of different parameters such as molecular weight,
tie line length, volume ration and neutral salt (NaCI) were
studied employing PEG 4000/potassium phosphate system
and PEG 4000/sodium sulphate system to select the best
system among the two.
Zhu et al. [39] performed the extraction of c-
phycocyanin from fresh Spirulina platensis by deploying a
species of non-pathogenic nitrogen-fixing bacteria, namely,
Klebsiella pneumoniae. The algal slurry was neither washed
nor centrifuged; the bacterial culture was poured into the
slurry, the vessel sealed, and crude C-PC extracted after
about 24 hrs. The extraction was clean and efficient, and the
purity and concentration of C-PC proved to be of adequate
quality. Asha Parmer et al. [40] extracted, purified and
characterized the C-phycocyanin using novel method based
on filtration and single step chromatography. The protein
was extracted by repeated freeze-thaw cycles and the crude
extract was filtered and concentrated in stirred ultra
filtration cell (UFC). The UFC concentration was then
subjected to a single Ion exchange chromatographic step. A
purity ratio of 4.15 was achieved from a starting value of
1.05. The recovery efficiency of C-phycocyanin from crude
extract was 63.50%. The purity was checked by
electrophoresis and UV spectroscopy.
Zhang et al. [33] obtained C-phycocyanin under
the best operational conditions for high C-phycocyanin
recovery and purity using the precipitation technique. Crude
C-phycocyanin from Spirulina platensis was used. The best
purification condition was ammonium sulfate fractionation
at 0-20%/20-50%, in relation to a resuspension
volume/initial volume of 0.52 in a 7.0 pH buffer. Under
these conditions, in an one-step purification only, the purity
increased 70% compared to the initial extract, with an
83.8% recovery.
MEDICINAL USES OF Spirulina platensis
Studies have shown that Spirulina consumption
during 4 weeks reduces serum cholesterol levels in human
beings by 4.5% [41] and significantly reduces body weight.
Spirulina extract induces the tumor necrosis factor in
macrophages, suggesting a possible tumor destruction
mechanism (Shklar and Schwartz, 1988) [42]. An extract of
sulfated polysaccharides, called Calcium-Spirulina (Ca-SP),
made up of rhamnose, ribose, mannose, fructose, galactose,
xylose, glucose, glucuronic acid, galacturonic acid, and
calcium sulfate, obtained from Spirulina, showed activity
against HIV, Herpes Simplex Virus, Human
Cytomegalovirus, Influenza A Virus, Mumps Virus and
Measles Virus. Cell extract of Spirulina maxima has shown
antimicrobial activity against Bacillus subtilis,
Streptococcus aureus, Saccharomyces cerevisiae, and
Candida albicans. Spirulina reduces: hepatic damage due to
drug abuse and heavy metal exposure, inflammatory
response, cells degeneration, anaphylactic reaction.
Spirulina contains vitamin A, important in preventing eye
diseases; iron and vitamin B12, useful in treating hypoferric
anemia and pernicious anemia, respectively; γ-linolenic
acid, appropriate in treatment of atopic child eczema
therapy; to alleviate premenstrual syndrome, and in immune
system stimulation [16]. Spirulina has been studied as an
animal cell-growth stimulant and in the treatment of
residual waters using alginate [18].
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platensis using factorial design. Bioresource Technology, 98, 2006, 1629-1634.
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