Vol 3 Issue 1 Bio Sci Article No

67 | P a g e International Standard Serial Number (ISSN): 2319-8141
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International J ournal of Universal Pharmacy and Bio Sciences 3(1): J anuary-February 2014
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Bio Sciences RESEARCH ARTICLE!!!

PRODUCTION, CHARACTERIZATION AND ANTIMICROBIAL ACTIVITY
OF MONASCORUBRAMINE FROM MONASCUS PURPUREUS
A. Avila Jerley
*
and Nisha Jayasankar
Post Graduate and Research Department of Microbiology, Dr. N.G.P Arts and Science College,
Coimbatore- 48.

KEYWORDS:

Monascus purpureus, red
yeast rice, wound
pathogen, antibacterial,
antifungal, pigment.
For Correspondence:
A. Avila Jerley*
Address: Post Graduate
and Research Department
of Microbiology,
Dr. N.G.P Arts and
Science College,
Coimbatore- 48.
Tamil Nadu, India.
Mail id:
ajerly4@gmail.com
Mobile No:
09943643370.

ABSTRACT
Many filamentous fungi synthesize various natural pigments which
have ecological functions and are of value to the producers. Among
them the fungi Monascus purpures, unique in nature produces
secondary metabolites like antihypercholestrol agent- Monacolin K,
hypotensive agent- amino butyric acid (GABA), antioxidants-
dimerumic acid and six pigments of polyketide structure. In this paper
the studies were focussed on Solid-state fermentation which was carried
out using rice as a substrate for the production of pigments using a
fungal culture of Monascus purpureus MTCC 1090. The pigment
complex was extracted with solvent system and purified by column
chromatography and TLC to obtain the red pigment fraction,
monascorubramine. The spectrophotometric characterization showed
max at 500 nm. FT-IR analysis was done to study the presence of their
functional groups which confirms their structure. Further the
antimicrobial effect of this pigment was investigated against wound
pathogen of some bacterial and fungal strains. They exhibited their
antibacterial activity against microorganismof the genera Bacillus,
Pseudomonas and E.coli and antifungal against Aspergillus and
Pencillium sps. The observation of bacteriostatic and antifungal effects
has lead to the consideration that the pigments of Monascus purpureus
has a therapeutical property.

INTRODUCTION:
The present trend throughout the world is shifting towards the use of eco-friendly and biodegradable
materials, the demand for natural pigment is increasing, so now microbial pigment is gaining its
popularity among the researchers. Microorganisms are known to produce a variety of pigments;
therefore they are promising source of food colorants (Ali, 2011 and Ahmad, et al., 2012).Some of
the most important natural pigments are carotenoids, flavonoids, tetrapyrroles and some xanthophylls
as astaxanthin. Many species of higher fungi (Basidiomycetes and Ascomycetes) produce brightly
colour fruiting bodies in range of pink, red, orange, yellow, purple, olive green, and grey which is
one of the important characteristics in the identification of some species like Agaricales sp. The
fungal mycelium gives rise to many secondary metablolites. The pigments produced by filamentous
fungi include quinines such as anthraquinones and naptha quinones, dihydroxy naphthalene melanin
and flavin compounds such as riboflavin (Duran et al., 2002).
The genus Monascus includes important species like Monascus purpures, Monascus ruber &
Monascus pilosus, which produces secondary metabolite like antihypercholestrol agent Monacolin
K, hypotensive agent- amino butyric acid (GABA), antioxidants-dimerumic acid and some
antibacterial compounds of polyketide structure. Among them the fungi Monascus purpures is
unique in nature which produces red pigment which is of high demand in substituting nitrites in food
products. Monascus purpureus belongs to the family Monascaceae and class Ascomycota whose
characteristic features is ability to produce secondary metabolites with strong yellow, orange or red
pigmentation. It can be easily distinguished by its ascospores. Monascus sp produces a complex
mixture of six chemically defined colored compounds, they are orange pigment: rubropunctatin
(C
21
H
26
O
5
) and monascorubrin (C
23
H
26
O
5
), yellow pigment: monascin (C
21
H
26
O
5
) and
ankaflavin(C
23
H
30
O
5
), red pigment: rubropunctamine (C
21
H
23
No
4
) and monascoruramine
(C
23
H
27
No
4
) (Wang et al., 2004).Monascus pigments are group of fungal secondary metabolites
called azaphilones, which have similar molecular structure as well as chemical properties. Monascus
purpureus is used for fermenting red yeast rice. Monascus purpureus produces intense red pigment
as well as metabolic by products when cultivated on cooked glutinous and non glutinous rice. The
fermentation and pigment production was carried out in solid culture and liquid culture with various
starchy substrates (Chairote et al., 2009). The pigment production varies greatly with the species and
cultivation conditions.This Red yeast rice (RYR) was considered as good medicine for digestion and
revitalization in ancient Chinese pharmacopoeia of medicinal food and herb. The pigments and other
bioactive substances have wide range of biological and therapeutic benefits, including anti-

carcinogenic, anti-oxidative, and hypolipidemic activities., RYR are used to reduce the risks of
oxidative stress in diabetes mellitus and hyperlipidemia (Rajasekaran and Kalaivani, 2011).
This work focuses the natural pigment production by cultivating Monascus purpureus under SSF.
The pigment production were measured, purified and analysed for the presence of functional groups
to elucidate the structure. Finally, the pigments were tested for its antimicrobial activity against
various pathogens.
Materials and Methods:
Collection of Fungal Strain: Fungal strain Monascus purpureus MTCC-1090 (Microbial Type
Culture Collection) was obtained from the Institute of Microbial Technology, Chandigargh.
Cultural Maintenance of Fungal Strain:
The fungal strain was enriched and maintained in Potato Dextrose Agar medium (PDA) at room
temperature for 10 days. After full growth the spore and their hyphae structures were identified
microscopically and it was subcultured every 30 days.
Microscopic Observations of Fungal Strain
Slide Culture Technique: Slide culture technique was used to identify the spore and hyphae
structures of fungal strain by lactophenol cotton blue staining method. Aseptically a circular sheet of
filter paper was placed on the bottom of the petri dish. The filter paper was moistened with sterile
water and a sterile glass slide was placed on it. A 5mm square block was cut from the aseptically
prepared PDA medium using a sterile scalpel and placed on to the sterile petri dish which was
already prepared. The fungal mycelium was placed on the four corners of the medium. Then a sterile
cover slip was placed over the agar blocks. A small sterile wet cotton balls was kept inside the
petridish to maintain the moisture condition and incubated at room temperature for 3- 4 days. After
incubation, the hyphal structures were visualized by lactophenol cotton blue staining.
Preparation of Monascus Fermented Rice
Indian rice was obtained from local market of Madurai, Tamil Nadu ,India and tested for its
use as a substrate for preparation of Monascus fermented rice with high concentration of
statin related compounds. Hundred grams of rice was soaked in distilled water (1L) at room
temperature for 8 hours and excess water was removed after soaking. The soaked rice was
autoclaved for 20 min at 121
C in an autoclave. Substrates were cooled and then inoculated with 10

days old precultured Monascus purpureus inoculam. The inoculated rice was incubated at 30
C for 2
weeks. The end products were dried in the oven at 65
C for 6 hours to obtain the dried Monascus

Fermented Rice (MFR).The dried MFR was pulverinzed and stored for further analysis. The yield of

MFR was evaluated as percentage yield. The percentage yield were obtained by using the following
formula
Weight of dried end product
Percentage of yield = --------------------------------------- x 100
Weight of steamed Rice used

Extraction of Red Pigment from MFR
The Red pigment was extracted with 95% of ethanol with the proportion of 5ml ethanol/ gm of dry
MFR. This mixture was kept on a rotary shaker at 200rpm for one hour. Then it was centrifuged for
15 minutes at 10,000rpm and the supernatant was collected for pigment analysis.
Purification of Pigments using Column Chromatography
Silica gel of 60-120 mesh size adsorbent used for the purification of the pigment was packed in a
glass column of dimension 10 x 210 mm . The column was packed with silica gel by slurry method.
The column was pre eluted using the solvent chloroform: ethanol (9:1, v/v). 0.1gm of MFR was
loaded and eluted using the same solvents. The different fractions of the pigments were collected for
further analysis.
Determination of Red Pigment
Pigment estimation was done by method described by Tseng, (2000) in which the optical density at
its absorbance maxima were expressed as the concentration of pigment produced. The absorbance
maxima of the pigment extract was measured by spectral analysis at 510 nm using double beam
spectrophotometer (Shimadzu, UV), taking into consideration the dilution factor of the sample OD at
its maxima, per gram of dry fermented matter (Johns and Stuart, 1991).
AU/gm of Pigment = O.D x Dilution x Volume of extracts/ Amount of sample (ml)
Analysis by Thin layer Chromatography
The microbial pigment extracts were subjected to thin layer chromatography (TLC) to separate the
bioactive compounds. The TLC plates were prepared using silica gel G and distilled water. Silica gel
G (20g) was added to 40ml of distilled water and thick slurry was made. Using a microcapillary tube,
a small drop of pigment extract of selected microorganism was placed on the TLC plate, 3cm above
the bottom. The spot was allowed to dry and the TLC plate was placed into the TLC chamber which
was saturated with Benzene:methanol:water (30:10:10) solvent mixture. When the solvent reached 2
cm below the top, the plates were taken out of the chamber and detected for the spots and Rf value
was calculated.


Chemical Structural Analysis of Purified Pigments
The chemical structure of the purified pigment was analyzed by UV-Visible Spectrophotometer and
Infrared Analysis
Spectral Analysis by UV- Spectroscopy
A UV-Visible spectrophotometric analysis was done to detect the presence of UV-absorbing
compounds in all the pigments using Shimadzu (Japan) model UV- 1700 series Spectrophotometer.
The spectral range for Monascorubramine is around 350 to 500nm.
Spectral Analysis by Fourier Transfer Infrared Spectral (FTIR)
IR spectral analysis of three isolated pigments was done by keeping the samples in vacuum
desiccators over solid KOH for 48 hours and then IR spectral Analysis was done with 1mg sample in
a Fourier transfer Infrared spectrophotometer (Shimadzu, japan).
Antimicrobial Activity of pigment against Pathogens
Test Microorganisms
Seven bacterial strains and the four fungal strains used in the present study were the clinical isolates
obtained from Aravind Eye Hospitals, Madurai. The bacteria used were Escherichia coli,
Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus
epidermis,Salmonella typhi and Acenitobacter sp.
The fungal strains used were Aspergillus niger, Alternaria sp,Penicillum sp,Fusarium oxysporum.
Antibacterial Assay
The effect of fungal pigment extracts on the several bacterial strains was assayed by Agar well
diffusion method and by Disc diffusion method. The plates were then incubated at 37C for 24
hours. The antibacterial activity was assayed by measuring the diameter of the inhibit ion zone
formed around the well.
Antifungal Assay
The Antifungal activity was analyzed by preparing the experimental PDA plates supplemented with
1% of fungal pigment and only PDA plates served as control dish. The antifungal properties were
estimated against several fungal strains by inoculating with 2l suspension of conidiophores in the
central zone of the Petri dishes. After incubation the diameter of the cultures were measured and
compared with control. Finally the incubation ratio was calculated by the formula
Inhibition ratio (%) = C-E/C x 100
where C is diameter of the mold in control plate and E is the diameter of the mold in experimental
Plate.


Result and Discussion
Results
Cultural & Microscopic Examination of Monascus purpureus:
The morphocolonical aspects of Monascus purpureus was characterized for its pigment production
on PDA medium. Monascus purpureus formed a typical fungal colony with white fluffy growth,
with red or orange pigmentation at the centre (Figure 1 a, b) and after 15 days of incubation, there
was improved growth with slight pink to red coloured pigmented colonies which were orange on the
ventral view (Figure 1c, d). Throughout the growth period the plates were maintained at 30
o
C.
On microscopic observations by lactophenol cotton blue staining method showed (Figure 2,e) the
cells appeared as septate hypha with hyaline walls of 3-5 m diameters. This hypha has sexuate
asca with ascospores, closed in ascocarp and posses asexuate conidiospore. The ascocarpe were
globular in shape containing 20-70 m of diameter of ascospore. This ascospore was oval ellipsoidal
shape with smooth hyaline walls (5-6 x 3-4 m). In asexuate reproduction, the conidiospores were
chained with basipetal succession having conidia of 7.5 x 5-6.2 m in dimensions, present with in
hyaline, subglobose, thick and smooth walled structure.
Microbial Conversion of rice to Monascus Fermented Rice
Generally, Pigment production was carried out using submerged fermentations. However solid state
fermentation and solid state fermentation system appeared to be promising. Here in this study rice
was used as substrate, a potential source of carbohydrates and also one of the best carbon source for
the pigment production. (Caralho et al., 2003). Monascus purpureus turned white rice to reddish
coloured within 3 days of fermentation, a sign of pigment production and continued to diffuse and
accumulated throughout the fermentation period, to its highest level at 10
th
day of incubation (Figure
2,a,b).Temperature plays a major role in cell synthesis and the red pigment was maximum at 30
o
C
(Sandipan et al., 2009).
The percentage of yield of MFR after 2 week cultivation was determined and it was found to be
47.15%, this was similar with the results of Chairote et al., (2007). The red rice production varied
from pale red to dark red depending up on the kind of rice used and the rice was broken and stucked
together, the texture was not so hard with pleasant odour.
Extraction, Purification and Estimation of Red pigment from Monascus purpureus
The selection of extraction methods depends on the cell structure, fermentation condition, substrates
and solvents used. The mixture of pigments synthesized by the Monascus mold grown on rice culture
medium was extracted using 95% ethyl alcohol (Figure 3).The extracted red powdered pigment was
further purified using silica gel column chromatography and the purified fractions were estimated for

its pigment content. The contents of red pigment in ethanolic extracts was measured by UV spectral
analysis and the as absorbance unit per gram was found to be 2.41 AU/g.
FIGURE 1:
STEPWISE GROWTH OF MONASCUS PURPUREUS ON POTATO DEXTROSE AGAR

(a) Growth after 3 days (b) Formation of red pigmentation

(c) Dorsal View of Monascus purpureus (d) Ventral View of Monascus purpureus

MONASCUS MYCELIUM AND ITS SPORES ON LACTOPHENOL COTTON BLUE STAINING
(e)


FIGURE 2:
PREPARATION OF MONASCUS FERMENTED RICE BY SOLID SUBSTRATE
FERMENTATION
(a) (b)

FIGURE 3
EXTRACTION OF RED PIGMENT FROM MONASCUS PURPUREUS

Charcterization of Microbial Pigments
Analysis by Thin layer Chromatography
The plate developed in benzene: methanol: chloroform showed visible bands of red orange and
yellow (Figure 4). The various pigments present in Monascus purpureus were distinguished based on
their Rf values such as 0.23, 0.73, 0.81 that corresponds to the red, orange and yellow pigments.
These bands were found to be monascorubramine and monascin of Monascus purpureus.
(Vidyalakshmi et al., 2009).

Figure 4: Chromatogram of biopigment from MonascusPurpureus

Spectroscopic Measurements
The absorbance properties of the microbial pigments facilitate both qualitative and quantitative
analysis. Here in this study the UV-Visible absorption spectra of microbial pigments are of immense
importance, since they aid in a great deal in determining the structure of the microbial pigments.
The Red pigment from Monascus purpureus was completely extracted in ethanol and this purified
pigment showed max at 510nm.The absorbance of the red pigment was presented in the Figure 5.
This was identical to the findings of Camelia &Mariana (2010) and was confirmed as
monascorubaramine.
Figure 5: Spectra of Fungal Pigment Monascorubramine

Fourier Transfer Infrared Spectral (FTIR) Analysis
IR spectral analysis of three isolated pigments was done and their spectrum has the following
functional groups.
The FTIR Spectrum of the red pigment (3500-350cm
-1
) is shown in the Figure 6 and the Table 1
represents the functional groups present in the compounds.

Table 1. Functional Groups of Red Pigment from Monascus purpureus
Wavenumber (cm
-1
) Functional groups
802.33 Aromatic C-H bend(meta group)
881.41 Aromatic C-H bend(para group)
1380.94 Alkanes with CH
3
bending
1414.65 C-H bending in alkene group
1656.74 C=O stretching in amide group
2891.1, 2927.74 C-H stretching in CH
2
groups
2974.03 C-H stretching in CH
3
groups
3346.27 Strong band for N-H stretching in secondary amine.

From the IR spectrum there was a strong peak for amine and aromatic groups which confirms that
this red pigment is monascorubramine (Sandipan et al., 2009). The FTIR Spectrum of the yellow
pigment (3500-500cm
-1
) is shown in the Figure 5.1.5.2c and the Table 5.1.5.2c represents the
functional groups present in the compounds.
FIGURE 6. IR Spectra of Fungal Pigment Monascorubramine

Potential Antimicrobial Therapy and their Medicinal Properties
Antibacterial Assay
The in vitro antibacterial activity of the microbial pigments was tested against the several test
organisms by well and disc diffusion methods and ampicillin was used as control (10g/ml). The
antibacterial activity potentials were assessed by the presence or absence of inhibition zone in
diameters.
The red pigment monascorubramine from Monascus purpureus showed highest antibacterial activity
against Pseudomonas aeruginosa, with inhibition zone of 24 mm and 21 mm followed by

Staphylococcus aureus of 17 mm and 14 mm and Streptococcus epidermis of 14 mm and 11 mm
respectively Figure 7,8,a,b,c. In both the diffusion methods, the highest antibacterial activity was
observed in same test organism with slight difference in measurements of inhibition zones. The
maximum antibacterial activity was observed in Gram negative organism than the Gram positive
organisms.
FIGURE 7. ANTIBACTERIAL ACTIVITY OF FUNGAL BIOPIGMENT FROM
MONASCUS PURPUREUS

FIGURE 8
DETERMINATION OF ANTIBACTERIAL ACTIVITY AGAINST VARIOUS PATHOGENS - DISC
DIFFUSION METHOD

(a) Z.O.I of Red pigment against (b) Z.O.I of Red pigment against E.coli
Pseudomonas aeuroginosa
0
5
10
15
20
25
30
Z
o
n
e

o
f

I
n
h
i
b
i
t
i
o
n

i
n

m
m
Test organisms
Disc Diffusion Method Well Diffusion method

(c) Z.O.I of red pigment against
Pseudomonas aeuroginosa
Antifungal Assay
Antifungal activity was estimated using four fungal strains as test organisms, cultivated on potato
dextrose agar medium (control plates) and on the same medium supplemented with 1% of microbial
pigments(Experimental plates).The diameter of the colonies were measured and the inhibition ratios
were calculated for each fungal strain. The red pigment monascorubarmine from the fungi showed
maximum of 64% inhibition in Penicillium sp and minimum of 31% in Alternaria sp.(Figure 9)
Moreover in the fungal strain grown in medium with microbial pigments showed change in fungal
colonies with less hyphal growth and diminished spore formations.
FIGURE 9
DETERMINATION OF ANTIFUNGAL ACTIVITY OF BIOPIGMENT FROM
MONASCUS PURPUREUS

0
10
20
30
40
50
60
70
Aspergillus sp Penicillum sp Alternaria sp
Fusarium sp
Z
o
n
e

o
f

I
n
h
i
b
i
t
i
o
n

i
n

P
e
r
c
e
n
t
a
g
e
Test Organisms

Discussion:
There is a growing interest in natural colourants which provides additional physiological benefits for
human health, other than the basic nutritional and energetic requirements. Biocolourants are mainly
derived from pigments and pigmentation is one of the important characteristic features of many
species of microorganisms. Some species of bacteria, fungi and algae produce pigments, which are
light absorbing compounds that are responsible for the colours that organisms display. Monascus
Species are known to produce well known water soluble azophilone pigments. Among the various
strains of Monascus sp, Monascus purpureus is used for the study. This forms orange to red coloured
fungal colony on PDA medium. The inorganic nitrogen compounds present in the medium enhance
the yield of red pigments. The pigment production is high and they seccrete red coloration into the
medium, when there is reaction between NH
2
group and inorganic compounds of the medium (Kaur,
et al., 2009). The Microscopic slides revealed the sexuate asca with ascopores and asexuate hypha
with conidiospores in basipetal arrangements.
High amount of red pigment production was observed in solid state fermentation where, cooked
autoclaved rice act as substrate. Due to the presence of high amylopectin and amylase there was
higher degree of starch hydrolysis, which plays a major role in microbial conversion of rice for the
pigment production. The substrate with high carbohydrate and moisture content could serve as better
fermentation media for Monascus purpureus (Carvalho et al., 2003). The levels of oxygen and CO
2

environment also significantly affect the pigment production. Hence the batch culture production of
red pigment by Monascus purpureus gives good yield of MFR. The growth of Monascus purpureus
was measured on the dry weight basis and pigment production. There was more or less steady
increase in biomass and pigment colouration as incubation period increases. The pigment production
was observed on 3
rd
day of incubation and continued to accumulate throughout the fermentation
period.Mycelial growth increased and reached maximum on 10
th
day at 30
o
C (Rashmi and
Padmavathi, 2011).The MFR yield was 47.15% after two weeks of cultivation and contents of red
pigment in 95% ethanol extracts was 2.41 Au/g (Chairote et al., 2007).
Characterization of Microbial Pigments
Thin layer chromatographic analysis of the pigments revealed the presence of their functional groups
with the specific Rf value. The separation of these biopigments was based on the differential affinity
of the compounds and their respective stationary and mobile phase. The extraction of red pigment
from Monascus purpureus gave two major spots on TLC with the Rf values same as the findings of
Vidyalakshmi et al., 2009.In this study orange and yellow spots were found to be maximum because

the red colour monascorubramine pigment usually mixed with rubropunctatin and gives orange
colour, when it is oxidized they form yellow colour monascin (Maria et al., 2004).
The qualitative measurements of the pigments and colourants are specifically done by observing the
max by UV-Visible spectrophotometry, the standard technique for quantifying the colourants that
is solublized in their respective solvents. The UV spectral data of a red pigment showed maximum
absorbance around 510nm.Many reports also indicates that absorbance of red pigment from
Monascus purpureus is ranges from 400- 500nm (Babitha et al., 2007).The red colour is due to the
presence of monascorubramine. The amine compounds play a main role in derivatization of red
coloured compounds by ring opening and shift rearrangements reaction (Chairote, et al., 2007).
FTIR is one of the most widely used methods to identify the chemical constituents and elucidate the
compounds structure to identify medicine in Pharmacopeia of many countries (Margarita, et al.,
2000). It is an accepted tool for the characterization of biomolecules in drug discovery research by
development of high- throughput screening and combinatorial chemistry.In this present work FTIR
spectroscopy operates in mid infrared region of 4000-400 cm
-1
which has been proved to be powerful
tool for quantitative analysis of microbial pigments. The IR absorption band of red pigments
indicates the presence of amine (N-H stretching) and for aromatic groups which confirms the
monascorubramine structure (Sandipan et al., 2009).
Among the vast number of methodologies used to relate the antimicrobial activities of microbial
pigments, the most popular methods employed are disc and well diffusion methods. In the present
study microbial pigments from selected microorganisms inhibit the growth of other microorganisms
or reduce the growth in the medium by the production of their secondary metabolites. These
metabolites produced indirectly when there is change in pH, osmotic pressure and surface tension or
directly by producing toxic components, antibiotics and other antimicrobial agents. This antagonistic
interaction among the microbiota can be used as biological attribution in food and pharmaceutical
industries.
The red pigments from Monascus purpureus showed activity against both Gram positive and Gram
negative bacteria. These results shows that pigment derivatives with phenyl ring have high
antibacterial activity. The hydrophobicity of pigment derivatives and the amount of pigment
absorbed by the cell surface also influence the antibacterial activity. Pseudomonas aeruginosa
showed maximum activity and this fact agrees with the reports of Jung et al,2003.
The microbial pigments posses antifungal activity and many fungicides exhibit lower potency under
field condition and are ineffective due to the development of resistance towards many synthetic
chemicals. These microbial pigments naturally have some defensive chemical compounds such as

alkaloids, phenolic compounds, antibiotics, terpenes, proteins etc., They diffuse into the microbial
cell membrane and penetrate into the cell where they interfere the metabolic pathway by hindering
the synthesis of glucan, chitin,ergosterol proteins and glucosamine in fungi causing inhibition in
growth of fungal colonies (Marino et al., 2001) and the monascorubramine are effective in inhibiting
the hyphal growth of all four fungal strains.
The toxicity problems caused by synthetic pigments to the environment and human health have
created an interest towards the natural pigments. Nowadays, complementary and alternative
medicine is widely available throughout the world. Several microorganisms have the capacity to
produce multifaceted secondary metabolites like pigments, which can play various roles as
biocolourants, antioxidants, antimicrobial agents and potent pharmacological drug against diseases.
From the above results summarized in this present study, the microbial pigments monascorubramine
isolated from fungi Monascus purpureus, has remarkable biological effects with therapeutic
applications in clinical and biomedical research on further level. These qualities make the microbial
pigment to use as therapeutic molecules in human health concern and eco-friendly molecules.
Currently, Commercial production of microbial pigments from microorganisms competes mainly
with synthetic manufactures by chemical synthesis. Thus, more detailed studies and scientific
investigations are needed to assess the real potential and availability of microorganisms to produce
natural pigments. Thus the present study will help to understand further therapeutic application of the
microbial pigments for food additives or pharmaceutical agents by animal model studies.
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C in an autoclave. Substrates were cooled and then inoculated with 10

C for 6 hours to obtain the dried Monascus

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