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Aroumougame, IJPSR, 2021; Vol. 12(4): 2370-2379. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 2370
IJPSR (2021), Volume 12, Issue 4 (Research Article)
Received on 03 April 2020; received in revised form, 17 August 2020; accepted, 05 November 2020; published 01 April 2021
STUDIES ON IN-VITRO ANTICANCER AND ANTIOXIDANT PROPERTIES FROM MARINE
BACTERIAL PIGMENT ISOLATED FROM THE COASTAL AREA OF MARAKANAM (TN)
Sandanakirouchenane Aroumougame
Department of Microbiology, School of Life Sciences, Pondicherry University, Puducherry- 605014, India.
ABSTRACT: Marine bacteria have the potentiality to produce diverse
bioactive molecules such as pigment. Therefore, it needs to exploit and
identifying a novel type of pigment from marine bacteria for Industrial
applications. This study aimed to investigate the marine bacterial pigment
against antioxidant and anti-cancer properties; the marine bacteria producing
pigment were isolated from water samples collected at the coastal of
Marakkanam (TN), India. The isolates were screened out based on the
growth characteristics and performance of different media and the strain
designated as MB4, which was taken as further studies. The strain MB4
characterized by SEM analysis showed that coccoid cell morphology,
nonsporulating, Gram-positive with yellow pigmentation and positive for
MR-VP, catalase, lipase, acetoin production, and hemolysis. The cells were
able to tolerate 10 percent NaCl concentration and ability to grown pH 9.
The MB4 strain was shown a higher wave-number (1395.77) cm-1 against
Raman Intensity to identify pigment production. The methanolic extracted
pigment was produced at a maximum peak at 260 nm. The yellow-pigmented
crude extract checked for anti-cancer properties using the colon cancer cell
line (HCT15), the cell viability has been reduced after treatment of the extract
(25-500 µg ml-1) and also exhibits IC50 value of 255.58 ± 43.51 mg ml-1
antioxidant DPPH radical scavenging activity. Due to their yellow pigment
productions which have antioxidant activity and anti-cancer properties, this
could be a novel pigment-producing strain for biomedical and industrial
applications.
INTRODUCTION: Synthetic pigments made up
of heavy metals and petroleum compounds are
reported to be carcinogenic, allergic, induce
hyperactivity, toxic, and organ damage, which are
unsafe for both environment and human health 1.
Due to their low cost, increased stability, and wide
range of spectra, the synthetic pigments are widely
used 2.
QUICK RESPONSE CODE
DOI:
10.13040/IJPSR.0975-8232.12(4).2370-79
This article can be accessed online on
www.ijpsr.com
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.12(4).2370-79
In recent years, natural pigments are highly
demanding for the use of colouring agents in foods,
fabrics, feed, printing ink, and cosmetics, which is
nonpolluted, eco-friendly, and less cost.
Naturally occurring colourants are safe to use
produced by microflora and fauna, which are
nontoxic, noncarcinogenic and easily degradable 3.
Generally, microbial pigments are the micro-
organisms that produce colour, which is more
attractive nowadays due to easy methods available
for cultivation; pigments are highly stable and year-
round availability 4. An alternative source for
natural pigment produces from bacteria, fungi, and
microalgae 5-9.
Keywords:
MB4 strain, Yellow pigment, DPPH
activity, Anti-cancer properties,
Raman spectroscopy, Marine water
Correspondence to Author:
Sandanakirouchenane Aroumougame
Research Scholar,
Department of Microbiology,
School of Life Sciences, Pondicherry
University, Puducherry - 605014,
India.
E-mail: sandana03@yahoo.co.in
Aroumougame, IJPSR, 2021; Vol. 12(4): 2370-2379. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 2371
Bacteria have great potential to produce various
bioproducts among the microorganisms, and one of
the physiological characters produced by the
bacteria is pigment production 10. Biomedical and
pharmaceutical industries utilize marine natural
products, and the bioactive compounds are
produced by marine bacteria 11. Microorganisms
are a promising source of natural pigments like
carotenoids, flavins, chlorophyll, quinines, and
prodigiosin are the pigments produced by coloured
microorganisms found in the different
environmental niche. In survey 12, yellow (31.3 %),
orange (15.2%), brown (9.9%), and red or pink
(5.4%) were found to be microorganisms recovered
from marine sources. Various concentrations of
minerals, a wide range of temperature, and it
should be the ability to tolerate different pH is the
good qualities of pigment producers 5. Marine
organisms produce a variety of metabolites that are
treated against antitumor, antioxidant, and
antimicrobial activities.
In recent years, Raman spectroscopy is a popular
analytical tool applied for the study of
microorganisms, especially pigments and
biomolecules, through qualitative and quantitative
analysis. Strong Raman signals were exhibits when
applied to the microbial pigments for the
understanding of different pigments, even pure
cultures as well as extracted pigments 13, 14. It has
been used to monitor different types of pigments
present in the microorganisms directly through pure
cultures and environmental samples 15.
Antioxidants may reduce the risk of diseases,
particularly heart and cancer, in the health sector. It
can absorb free radicals that may oxidize lipid or
DNA, proteins, nucleic acids and cause
degenerative disease 16. Many other findings
reported that the β-carotene and other carotenoid
extracted pigments have antioxidant activities 17.
Antioxidant able to donate a hydrogen atom
reduces DPPH as a result of colour loss which
determines the scavenging capacity of a molecule
18. The carotenoid pigment producer Planococcus
sp. TRC1 was showed appreciable antioxidant
activity leading to pharmaceutical and food
applications 19.
The activation, viability, and proliferation of the
cells were measured quantitatively in calorimetric
for MTT Assay 20. The dehydrogenase enzymes
associated with the endoplasmatic reticulum and
mitochondria in the living cell convert MTT into a
purple-blue formation which is water-insoluble.
The viable cell number is directly proportionate to
product formation and inversely proportional to
cytotoxicity 21. In this study, we are investigating
the pigmented bacteria isolated from marine water
samples to evaluate the antioxidant and anti-cancer
properties of the extracted pigment.
MATERIALS AND METHODS:
Collection of MARINE WATER SAMPLE:
Samples collected from the marine water surface at
different sites along with the coastal areas of
Marakanam (TN), India. The marine water samples
from the sea surface were collected by using Teflon
plates dipped into water, lifted horizontally, and
scrapped off the adhering surface film till the
procedure repeated the total volume of 30 to 50 ml
water sample was collected. The samples were
stored at 4°C until the isolation was carried out
within 24 h 22. The salinity of the collected marine
water samples was determined by using the
protocol reported by 23.
Isolation of Yellow-Pigmented Bacterial Isolates
from Marine Water: Isolation of pigment-
producing colonies from collected seawater
samples using selective Zobell marine agar medium
24. The plates were incubated at 37°C for 48 h, the
colonies showing yellow, red, orange, and brown
pigmentation subcultured for purification.
Screening, Morphological and Biochemical
Characterisation of the Isolates: The bacterial
isolate was plated on Zobell marine agar, Luria
Bertani agar and Tryptic soy agar incubated at
37°C for three days. By using standard
Microbiological techniques, the different cultural
characteristics, cell morphological parameters, and
Gram's reaction were studied to the bacterial cells.
Scanning Electron Microscopic (SEM) Analysis
for MB4 Isolate: The yellow-pigmented bacterial
culture isolate was grown on Luria Bertani broth
were centrifuged, phosphate buffer saline (pH 7.0)
used to wash for thrice to remove salts,
glutaraldehyde (2%) solution was fixed with
sample and allowed for alcoholic dehydration at 6-
12 h. The dehydrated sample was prepared and
analysed on SEM 25.
Aroumougame, IJPSR, 2021; Vol. 12(4): 2370-2379. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 2372
Biochemical Characterisation of the Pigmented
Isolate: The biochemical characteristics of the
yellow-pigmented bacterial MB4 isolate.
Kovac's Oxidase Test: By using a sterile
toothpick, a well-isolated colony of MB4 was
picked and thoroughly rubs into an area of the
moist test disc impregnated with oxidase reagent.
After 30 sec, the inoculated area was observed for
colour change. A bluish-purple colour indicated a
positive reaction 26.
Catalase Test: Cultures grown on MB4 slants for
24 to 48 h flooded with 0.5 ml of 3 per cent
hydrogen peroxide. Rapid effervescence shows a
positive result for catalase activity 27.
Methyl Red-Voges Proskauer Test (MR-VP
test): Methyl Red-Voges Proskauer tests were used
to differentiate acid producers from those
producing a neutral product, acetoin. The isolate
was inoculated in 5 ml MRVP tubes and incubated
at 35 °C for 48 h. Methyl red positive tubes were
observed by the change of the colour of the media
from yellow to red. Voges-Proskauer test was
recorded positive by the development of red colour
due to the addition of Barritt's reagent-I and
Barritt's reagent-II 28.
Lipase Activity: The isolate MB4 was inoculated
on egg-yolk agar and incubated at 37°C for 48 h.
The positive reaction of the lipase activity indicated
that the development of opalescent precipitates 29.
Urease Test: It was performed on 5 ml urea broth
in test tubes containing phenol red (pH 6.8) as the
pH indicator. The urea broth tubes inoculated with
isolates were incubated for 24 h. The positive tubes
were developed on red colour 30.
Assay of Phosphatase Activity: The loopful of
bacterial growth of strain was collected and
deposited on the surface of the tangible medium
containing the phosphatase substrate (para-
nitrophenyl phosphate) to a final concentration of 1
mg ml-1. The plates were then incubated at 37°C as
described by 31.
Coagulase and DNAse Production: It was
detected by the method using ethylenediamine-
tetraacetic acid (EDTA) treated coagulase plasma
by the formation of a clot after 1, 2, 4 or 24 h
recorded as positive. During the investigation, 0-1
% DNA (BDH) was added to this medium to
enhance the detection of DNase 32.
Physiological Characterisation of the MB4
Isolate: The physiological characteristics such as
pH and NaCl tolerance were determined. The effect
of pH was determined by the preparation of
nutrient broth with incremental pH values ranging
from 4 to 14. Strain MB4 was inoculated and
incubated for 48 h at 37°C, the growth in the
culture broth read at 620 nm. The pigments were
extracted and observed for maximum pigment
production 33. The inoculation of MB4 also
examined the effect of NaCl tolerance in nutrient
broth in different concentrations ranging from 6 to
12% of NaCl. The flasks were incubated at 37°C
for 48 h, and the results were observed for growth
and pigment production.
Extraction of pigments from bacterial isolates:
The yellow-pigmented MB4 isolate was grown in
LB broth for seven days kept in a rotary shaker at
160 rpm at 37°C. The broth was centrifuged at
8,000 rpm for 15 min, and the cells were harvested
and poured off the supernatant. The cell pellet was
rewashed with sterile distilled water centrifuged.
The pellets were resuspended with 5 ml methanol
and sonicate the mixture until all visible pigments
were extracted. The solvent mixtures were
centrifuged at 4,000 rpm for 15 min; the pigment
supernatant was separated and filtered through a
Whatman no. 1 filter paper and analyzed by
scanning the absorbance in the wavelength region
from 300-700 nm using a UV-VIS
spectrophotometer 34.
Raman Spectroscopy Analysis: The MB4 strain
was grown on LB agar medium after 48 h of
incubation and the pigmented pure culture to
determine the strong Raman signalling for Raman
spectroscopy 15.
Determination of DPPH Scavenging Assay: 35
was reported the method of determination of DPPH
radical scavenging activity. An aliquot of 0.5 ml of
the extracted pigment sample solution in methanol
was mixed with 2.5 ml of 0.5 mM methanolic
solution of DPPH. The sample mixture was shaken
vigorously and incubated for 30 min in the dark at
room temperature. The absorbance was measured
Aroumougame, IJPSR, 2021; Vol. 12(4): 2370-2379. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 2373
at 517 nm using a UV spectrophotometer. Ascorbic
acid was used as a positive control. DPPH free
radical scavenging ability (%) was calculated by
using the formula.
% of inhibition = absorbance of control – absorbance of
sample/absorbance of control ×100
Cell Culture and MTT Assay: The MB4
pigmented extract was carried out with MTT assay.
The HCT15 colon cancer cell line was plated
separately using 96 well plates with the
concentration of 1×104cells/well in DMEM media
with 10% fetal bovine serum and 1X Antibiotic
Antimycotic Solution in a CO2 incubator at 37°C
with 5% CO2. The 200 μl of 1X PBS cells were
washed, and then the cells were treated with
various concentrations of crude extract of the
pigmented compound in serum-free media and
incubated for 24 h. At the end of the treatment
period, the medium was aspirated from cells.
Prepare 0.5 mg ml-1 MTT mixed in 1X PBS and
incubated at 37°C for 4 h using CO2 incubator.
After the incubation period, the medium containing
MTT was discarded from the cells and washed
using 200 μl of PBS. The formed crystals were
dissolved with 100 μl of DMSO and thoroughly
mixed. The formazan dye turns to purple-blue
colour. The colour intensity was measured at 570
nm using a microplate reader 36.
Statistical Analysis: Statistical analysis was
performed with Mean and Standard Deviation (SD)
in excel, and all analyses were carried out in
triplicates.
RESULTS AND DISCUSSION:
Isolation of Pigmented Bacteria from Marine
Water: The bacteria producing pigment were
isolated from the marine water sample by using
Zobell Marine Agar media.
TABLE 1: SCREENING OF PIGMENT-PRODUCING
BACTERIAL ISOLATES
Location
Total no. of
isolates
Pigment producing
strains
Marakanam
coastal region
(12.1899° N,
79.9249° E)
MB1
MB2, MB4, MMB8
& SB2
MB2
MB3
MB4
MMB5
MMB8
MBS7
MBS8
SB2
SB10
Ten isolated strains were purified and named viz.,
MB1, MB2, MB3, MB4, MMB5, MMB8, MBS7,
MBS8, SB2 and SB10; the results were shown in
Table 1. Based on the growth performance and
pigment production, the strain MB4 was screened
for further studies.
Morphological Characters of Bacterial Isolates:
Pigment-producing bacteria are isolated from
marine samples, which are ubiquitous 37. A result
of Fig. 1, the isolated colony was round, smooth
with yellow-pigmented, non-motile, non-
sporulating, and gram-positive bacteria. 38 were
reported that the carotenoid-producing microbes
were isolated from the extreme environmental
niche. The sixty marine species of yellow (19),
orange (5), pink or salmon colour (5), brown (5)
and red (1) were described by 39. The marine
organism isolated in the present study was yellow-
colored. All bacterial pigmented isolates need not
be carotenogenic 40. Some pigmented bacteria from
marine are Alteromonas (yellow, violet) 41-43,
Flavobacterium (yellow) 44, Deleya, Marinomonas,
Pseudomonas and Shewanella (yellow) 45,
Erythrobacter (yellow) 46, Pseudoal-teromonas
(purple, red, or yellow pigments), Xanthomonas
(yellow) 47, Exiguobacterium (yellow, orange) 48, 34,
and Bacteroidetes (yellow, orange, pink or red) 49
were reported to possess antagonistic activities.
The SEM image of Fig. 2 was clearly shown that
the isolate belongs to Cocci with irregularly
arranged cells.
FIG. 1: GROWTH OF MB4 STRAIN ON LB AGAR MEDIUM
Biochemical Reaction of the Strain: The yellow-
pigmented bacterium from deep-sea sediment was
oxidase, DNase, methyl red, urease negative, and
catalase, Voges–Proskauer positive exhibited by
Croceicoccus marinus gen. nov., sp. nov., 50. The
present investigation of the MB4 strain was shown
in Table 2, which is positive for MR-VP, catalase,
lipase, acetoin production, and hemolysis and
negative for oxidase, coagulase, urease, and
phosphatase.
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International Journal of Pharmaceutical Sciences and Research 2374
TABLE 2: BIOCHEMICAL CHARACTERS OF THE
PIGMENTED STRAIN
S. no.
Biochemical reaction
MB4 strain
1
Oxidase
-
2
Catalase
+
3
Coagulase
-
4
Acetoin production
+
5
Phosphatase
-
6
Hemolysis
+
7
Urease
-
8
MR-VP
+
9
Lipase
+
Positive (+), Negative (-)
FIG. 2: SEM IMAGES OF PIGMENTED MARINE
BACTERIAL MB4 STRAIN
Physiological Characterisation of the Isolates:
Studies were reported the yellow-pigmented
Polaribacter butkevichii sp. nov. were grown in pH
between 7.6 and 8.2 isolated from marine water
samples 51. The MB4 strain exhibits maximum
growth at pH nine as compared to pH from 4 to 14
Fig. 3. The extreme halophiles were unable to grow
in the presence of NaCl less than 12%, and it can
also be able to grow in saturated NaCl, including
halobacteria and Halocooci. The isolate MB4 was
the ability to tolerate salt concentration of 10%
NaCl which was shown in Fig. 4. It has been
reported that the growth was exhibited up to 8.5%
NaCl concentration by P. balearica strain 52. The
KMM 1447T yellow-orange pigmented strain
isolated from marine ascidian was the ability to
grow in 8% NaCl 53.
FIG. 3: PIGMENTED MB4 STRAIN WAS GROWN AT
DIFFERENT pH
FIG. 4: THE BACTERIAL PIGMENTED MB4 STRAIN WAS
GROWN AT DIFFERENT NACL CONCENTRATION
Extraction of Yellow Pigment from the Strain
MB4: The isolated autotrophic cell cultures were
extracted yellow coloured pigments using methanol
solvent results much efficient in the quantification
was better than acetone 54. The yellow pigments
were separated from cell pellets by methanol
extraction from the bacterial broth Fig. 5. Based on
the absorption spectrum, the characterisation of
crude extracted yellow pigment exhibited a
maximum at 260 nm Fig. 6. As compared to water,
PBS or acetone extraction, pigments recovered
even better by using methanol extraction 34, 55, 56.
The lemon yellow colour pigment is extracted by
methanol from Halomonas aquamarina MB598,
also reported by 2.
FIG. 5: PELLETS ARE SEPARATED FROM THE
PIGMENTED MB4 STRAIN
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International Journal of Pharmaceutical Sciences and Research 2375
FIG. 6: METHANOLIC EXTRACTION OF YELLOW-
PIGMENTED BACTERIAL ISOLATES
Raman Spectroscopy Analysis for Pigmented
Strain: Raman spectroscopy, one of the highly
sophisticated instruments applied in Microbiology
research in recent years for the characterisation of
microbial pigments. The pure cultured MB4 strain
was analyzed by using Raman spectroscopy for
pigment production, and the strain was showed that
maximum wavenumber (1395.77) cm-1 against
Raman intensity Fig. 7. The excitation of electronic
absorption of the spectrum which produces a
specific enhancement of certain Raman intensity
(bands) correspondingly the portion of the
molecule electronic transition occurs indicates
moving of atoms in the chromophore when
vibration takes place 57, 58.
FIG. 7: RAMAN SPECTROSCOPY ANALYSIS OF
PIGMENTED MB4 BACTERIAL ISOLATE
Antioxidant Activity of Pigment Extract: Natural
products like pigments, frequently used to evaluate
antioxidant activity, the ascorbic acid, glutathione,
cysteine, and tocopherols, were used as reducing
agents which are decolorizing involving in DPPH
radicals 59.
The reducing activity of free radical owing to
antioxidant depending on the reduction of one
electron which exhibits scavenging of DPPH; as a
result development of antioxidant properties due to
reducing power 60. The anticarcinogenic and
antioxidants properties are present in carotenoid
pigments 61.
FIG. 8A: GROWTH PERFORMANCE OF A PIGMENTED EXTRACT OF MB4 AND ASCORBIC ACID
FIG. 8B: SCAVENGING ACTIVITY OF MB4 PIGMENTED EXTRACT AND ASCORBIC ACID
In-vitro, the potential of antioxidant activity of
yellow-pigmented crude extract (YPCE) was scavenged due to free radicals in 500 µg ml-1
concentrations, and the percentage of inhibition for
Aroumougame, IJPSR, 2021; Vol. 12(4): 2370-2379. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 2376
DPPH showed 68.30 percent Fig. 8A with IC50
255.58 ± 43.51 among different concentrations
from 25 to 500 µg ml-1 Fig. 8B.
The carotenoid pigment is the biological compound
available for human beings that have properties of
ulcer prevention 62. The DPPH free radical
scavenging activity was shown in different
concentrations by using the methanol extract 59.
The pigment production by halophilic bacteria and
their relation to radical scavenging property was
reported by 63. The yellow pigment was extracted
from Kocuria flava SIF3 discovered to have DPPH
radical scavenging activity with an IC50 value of
1.25 mg ml-1 in-vitro antioxidant assay 64.
Pigment Extract Evaluated for Anti-cancer
Activity: Although advanced techniques developed
in the treatment, prevention, and diagnosis of the
disease. Still, one of the most serious human health
problems in the world is cancer despite their
understanding of its biology, which causes severe
to mankind 65. Microbial pigments possess anti-
cancer activity, and prodigiosin pigment produces
cytotoxicity on U937 leukemia cells extracted from
Pseudoalteromonas sp. 1020R 66. The present
investigation was performed on a potential crude
extract of yellow pigment against colon cancer cell
line (HCT15), which showed that the different dose
concentrations of the methanolic fraction of MB4
isolated from seawater could largely inhibit cell
proliferation at a concentration of 500 μg ml-1 Fig.
9A and Fig. 9B and no cytotoxicity was detected
against the standard untreated cell. It was found to
be reported similar studies by 67.
25µg/ml Control 50µg/ml
100µg/ml 250µg/ml 500 µg/ml
FIG. 10: EFFECT OF MB4 STRAIN CRUDE EXTRACT ON COLON CANCER CELL LINE
Microbial pigments possess antimicrobial, anti-
inflammatory, anti-cancer, and antioxidant activities, which also act as colouring for various
industries like food processing and cosmetics 4. The
FIG. 9A: CELL PROLIFERATION OF CRUDE EXTRACT OF
MB4 STRAIN ON COLON CANCER (HCT15) CELL LINE
FIG. 9B: CELL VIABILITY OF MB4 PIGMENTED
EXTRACT ON COLON CANCER CELL LINE (HCT15)
Aroumougame, IJPSR, 2021; Vol. 12(4): 2370-2379. E-ISSN: 0975-8232; P-ISSN: 2320-5148
International Journal of Pharmaceutical Sciences and Research 2377
extracted crude pigment MB4 was checked against
the colon cancer cell line (HCT15) for cytotoxic
activity Fig. 10.
Many studies had reported that the inhibition of cell
cycle and apoptosis induced by microbial pigments
68, 69. It inferred that the crude pigment extract of
marine MB4 shown significant antioxidant and
potential anti-cancer activity against (HCT15) cell
lines of colon cancer. The cytotoxic activity against
cervical cancer cells (HeLa) and HepG2 were
demonstrated in yellow pigment from Streptomyces
griseoaurantiacus 70. The potential breast cancer
cell lines and lung cancer cells have experimented
with pigment carotenoid extracted from Kocuria
sp. QWT-12 71. The pigment was found to
exhibited inhibitory action against the growth of
human cancer cell lines, which develops anti-
cancer drugs from Salinicoccus sp. 72. The novel
compound from yellowish pigment produced by
Rhodococcus maris reduced the risk of breast
cancer 73. The Natrialba sp. M6 produce carotenoid
pigments that were effective against anti-cancer
and antiviral activities 74.
CONCLUSION: Bacterial pigments for different
therapeutics are an area of recent research interest.
The present study attempts to validate the anti-
cancer potential of bacterial pigment using colon
cancer cell line (HCT15) cells and antioxidant
properties. We used in-vitro cultured cells (HCT15)
to determine the anti-cancer activity of extracted
pigments from MB4 strain by using MTT assay for
measuring the cell viability, and our results depict a
dose-dependent by increasing concentration of the
extract which decreases in cell viability. The DPPH
radical scavenging activity revealed an increased
concentration of the extract with an IC50 value of
255.58 ± 43.51 mg ml-1. The anti-cancer and
antioxidant activity were induced by the extracted
marine bacterial pigments. The presence study
envisages the antioxidant and anti-cancer potential
of bacterial pigments isolated from marine water,
which can find applications in therapeutic
treatments. The organisms were isolated from
marine water, and yellow color pigment-producing
bacteria were used for pigment extraction.
Biochemical identification studies have confirmed
the MB4 organism to be gram-positive, Cocci with
a distinct yellow color, positive for MR-VP,
catalase, lipase, acetoin production, and hemolysis
which can tolerate 10 percent NaCl and it can also
grow pH at 9.
ACKNOWLEDGEMENT: We are thankful to the
Department of Microbiology, Pondicherry University,
Puducherry, for providing the facilities to carry out
this research work successfully.
DECLARATION OF INTEREST: The authors
declare no conflict of interest exists.
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Aroumougame S: Studies on in-vitro anticancer and antioxidant properties from marine bacterial pigment isolated from the coastal area of
Marakanam (TN). Int J Pharm Sci & Res 2021; 12(4): 2370-79. doi: 10.13040/IJPSR.0975-8232.12(4).2370-79.
