Detoxification of A Harmful Textile Azo

42 | P a g e International Standard Serial Number (ISSN): 2319-8141
International Journal of Universal Pharmacy and Bio Sciences 3(3): May-June 2014
INTERNATIONAL JOURNAL OF UNIVERSAL

PHARMACY AND BIO SCIENCES
IMPACT FACTOR 1.89***
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Bio Sciences Research Article ……!!!
DETOXIFICATION OF A HARMFUL TEXTILE AZO DYE SUNSET

YELLOW FCF BY MARINOBACTER SALSUGINIS SAY-3
1
Shertate R.S. and 2Thorat P.R.
1
Research Scholar, 2Associate Professor, P.G. Department of Microbiology and Research Center,
Shri Shivaji Mahavidyalaya, Barshi – 413411, Dist. - Solapur, MS, India.
KEYWORDS:
ABSTRACT
Azo dye, Detoxification, In the present study the textile azo dye degrading marine bacterium
Marinobacter salsuginis SAY-3 was isolated from the natural marine
GC-MS, Microbial
environment capable of degrading and detoxifying an azo dye Sunset
Toxicity. Yellow FCF at 5.0% NaCl under static condition in 24 hours. The
For Correspondence: maximum decolorization observed was 97.00% in nutrient medium
containing 5.0% NaCl at pH 8.0 when incubated at 300C temperature for
Shertate R.S.* 24 hours. Identification of the isolated strain SAY-3 was done by using
Address: Research biochemicals and 16S rRNA sequence analysis. The nucleotide
alignment and distance matrix showed the present strain with 93%
Scholar, P.G. Department
closest relation to Marinobacter salsuginis (EMBL Accession No.
of Microbiology and HF545597). The percent decolorization of the dye Sunset Yellow FCF
Research Center, was determined spectrophotometrically ( max 485nm).The efficiency of
decolorization was increased with the addition of carbon and nitrogen
Shri Shivaji
sources Viz. 1% Glucose, 1%Starch and 1% Yeast extract and was found
Mahavidyalaya, up to 98.00%, 98.55% and 100% respectively. After degradation
Barshi – 413411, products formed were analyzed by GC-MS technique and from this
Dist. - Solapur, MS, India. analysis it was found that the isolated organism degraded Sunset Yellow
FCF to the products having different molecular weights. The toxicity of
Email ID: the degraded products was checked by the microbial toxicity and was
rubinamicro13@gmail.com showed the degradation of Sunset Yellow FCF into non-toxic product by
Marinobacter salsuginis SAY-3. In percent COD reduction study, it was
observed that more than 90.00% COD was reduced. The present
research study highlights an efficient degradation and detoxification of
Sunset Yellow FCF by Marinobacter salsuginis SAY-3 from marine
environment.
Full Text Available On www.ijupbs.com

INTRODUCTION:
Sunset Yellow FCF is the textile azo dye with a structure disodium 6-hydroxy-5-{[4-
(sulfidotrioxidanyl) phenyl] diazenyl}-1, 4-dihydronaphthalene-2-sulfonate disodium salt. Azo dyes,
consist of one or more azo bond (-N=N-), account for 60-70% of all textile dyestuffs used1. The
inefficiency in dyeing processes has resulted in the discharge of 10-15% of unused dyestuff which is
directly entered in the wastewater2. The effluents having high levels of Biochemical Oxygen Demand
(BOD) and Chemical Oxygen Demand (COD) values are highly toxic to the biological life3. If such
type of colored effluent is directly discharged in the marine environment, water gets polluted which in
turn affect marine flora and fauna adversely. Although various physical-chemical methods have been
used to remove the color from effluents, but the major limitation of these methods is they are generally
expensive, produce large amounts of sludge and leads to the secondary pollution. Moreover, these
conventional treatment methods lead to the formation of some harmful side products. The presence of
the colorless aromatic amines in the aqueous ecosystems is of a serious environmental and health
concern; so that a complete removal of such compounds from aquatic ecosystem is required4. As a
better alternative, microbial biodegradation of dyes is now widely used5. Thus, bioremediation
overcome the limitations of the conventional techniques by the actual destructing many organic
contaminants at low cost. As a result, from the last two decades, bioremediation has becoming a
widespread technology that is considered for the cleaning up of a wide range of contaminants6. The
biological method of treatment is more economical and leads to less accumulation of relatively
harmless sludge. Moreover, biological treatment of dye bath effluents is ecofriendly. It converts dyes
into simpler inorganic compounds which are not lethal to aquatic life forms. The basic step in the
decolorization and degradation of azo dyes is breakdown of azo bonds, which is responsible for its
coloration, when such bond is broken the dye loses its color. Azo dyes are known to undergo reductive
cleavage whereas the resultant aromatic amines are metabolized under microaerophilic conditions7.
Some microorganisms like bacteria, fungi and algae, can degrade a wide range of dyes8. In the present
study, a potential marine bacterium Marinobacter salsuginis SAY-3 was isolated capable of completely
decolorize the textile azo dye Sunset Yellow FCF at 5.0% salinity in 24 hours.
Materials and Methods
Dyes and Chemicals
The textile dye Sunset Yellow FCF ( max-485nm) was purchased from Sigma-Aldrich (USA). All the
chemicals used were of highest purity and analytical grade.

Figure 1 Structure and properties of the dye Sunset Yellow FCF
Structure
HO
N
N
NaSO3 SO3Na
Properties
Molecular Formula = C16H10N2Na2O7S2
Formula Weight = 452
Composition = C(40.82%) H(2.51%)
N(6.35%) Na(10.42%)
O(25.37%) S(14.53%)
max = 485nm
Acclimatization and isolation of microorganisms

Saline Soil and Marine water was used as a source of microorganisms to isolate morphologically
distinct bacterial colonies capable for decolorizing the dye. The soil samples from nearby salterns
(Saltpan), areas nearby waste disposal sites of the textile industry, Marine water, sewage, sludge,
effluent treatment plants and compost as the source of microorganisms were collected and
homogenized properly. The soil samples were subjected to enrichment using nutrient broth amended
with increasing concentrations of NaCl (0.5% to 18.0%) and dye (250µg ml-1 to 10,000µg ml-1) with
incubation time of 24 hours at 30°C. Repeated transfers were carried out to isolate stable dye
decolorizing organism and the isolated organism was stored at 5°C for further use. The high
decolorizing bacteria were screened by performing a decolorization assay with the dye using UV-VIS
spectrophotometer (Systronics-106 model) at its respective max 485 nm and designated isolate SAY-
3 and used for further studies.
Optimization of Parameters
Effect of Static and Shaking Condition
The promising isolate was examined for their ability to decolorize the dye Sunset Yellow FCF in
nutrient broth medium containing (0.5%-18%) NaCl concentrations and 2000µg/ml concentration of
dye. The decolourization of Sunset Yellow FCF in static and shaking condition was studied.
Effect of Temperature
The optimization of temperature for the decolourization process was studied by inoculating the
promising organism in nutrient medium containing different NaCl concentrations and dye incubated at
different temperature ranges (250C, 300C, 370C, 400C and 500C) at pH 8.0.

Effect of pH
The optimization of for the decolourization process was also studied by inoculating the promising
organism in nutrient medium containing different NaCl concentrations and dye at different pH values
(5.0, 6.0, 7.0, 8.0, 9.0 and 11.0) keeping temperature constant 300C.
Effect of carbon and nitrogen sources on percent decolorization
The promising isolate was further supplemented with carbon and nitrogen sources Viz. 1% glucose,
1% starch and 1%yeast extract. The effect of these sources on the dye decolorization was also studied.
Determination of biodegradation activity
The bacterial strain which give maximum decolorization values was selected and used for further dye
decolorization experiments. The promising bacterium Marinobacter salsuginis SAY-3 was transferred
to fresh nutrient medium containing Sunset Yellow FCF (2000µg/ml) and 5.0% NaCl was incubated at
300C, under static condition for 24 hours. After 24 hours of incubation, aliquots (4ml) of the culture
media were withdrawn, centrifuged at 10,000rpm for 15 minutes in a cooling centrifuge at room
temperature to separate the bacterial cell mass. The supernatant was further used for the analysis of
decolorization and all the experiments were done in triplicates. Absorbance of the supernatant
withdrawn was measured at its absorbance maximum wavelength for the dye Sunset Yellow FCF
( max - 485nm) on spectrophotometer (Systronics-106). The percent dye decolorization was
calculated by measuring the difference between initial and final values of absorbance using the
following formula9.
Initial absorbance value-final absorbance value x100
% Decolorization = --------------------------------------------------------------------
Initial absorbance value
Percent Decolorization in
Percent Decolorization in Nutrient Broth and Half (½) Strength Nutrient Broth
The promising isolate SAY-3 was used to inoculate in 25ml nutrient broth containing 2000µg/ml
concentration of dye and 5.0% NaCl. Tube was then incubated at 300C for 24 hrs and observed for
decolorization of the dye. In addition, half strength nutrient broth (peptone – 0.5g, NaCl – 0.25g, Beef
Extract – 0.15g, Distilled Water – 100.0ml) was also used to test for the ability of isolate SAY-3 to
decolorize the dye Sunset Yellow FCF with same dye concentration and salinity.
Percent Decolorization by Cell Free Extract
The bacterial cells were harvested by centrifugation at 7,000 rpm, for 15 min, at 4°C and suspended in
50mM phosphate buffer pH 7.4. These cells (100 mg ml−1) were chilled properly and sonicated
(Sonics-vibracell-130 ultrasonic processor), keeping sonifier output at 40 amp and giving 7 strokes
each of 30 s, with time interval of 2 min at 4°C. The homogenate was then centrifuged at 8,000 rpm

for 15 min and the supernatant obtained was used as crude enzyme source. The supernatant containing
the crude enzyme was then added with 2000µg/ml concentration of dye solution and observed for dye
decolorization. The percent decolorization studies were monitored by using spectrophotometer.
Determination of Chemical Oxygen Demand (COD)
Percent COD reduction value of the dye decolorized by isolate SAY-3 was calculated by COD
analysis using K2Cr2O7 as a strong oxidizing agent under reflux conditions.
Extraction and analysis of biotransformed products
Biodegradation of Sunset Yellow FCF was monitored by GC-MS analysis. After decolorization, 100ml
sample was taken. Centrifugation was carried out at 10,000 rpm and the metabolites were extracted
from supernatant using equal volume of Dichloromethane. The treated Sunset Yellow FCF dye was
characterized by Gas Chromatography and Mass Spectroscopy.
Confirmation of Dye Degradation by GC-MS Technique
To study the products formed after degradation of azo dye Sunset Yellow FCF, decolorized samples
were analyzed by GCMS. The promising isolate was inoculated in 100mL of sterile nutrient broth
containing 2000 g/ml of dye Sunset Yellow FCF and 5.0% NaCl. The broth was then incubated at
ambient temperature for 24 hrs in separate flasks. The decolorized broth was then centrifuged at
10,000 rpm for 15 minutes in cooling centrifuge. Centrifugate was mixed with equal amount of
dichloromethane in separating funnel. Samples were shaken vigorously for 15 minutes and kept for 10-
15 minutes to separate solvent and liquid phases. After separation, liquid phase was discarded and
solvent phase allowed for partial evaporation. Partially evaporated samples were analyzed by GCMS
technique.
Detection of Degraded Products
The degraded products were detected by GC-MS technique.
Prediction of Dye Degradative Metabolic Pathway
The metabolic pathway of dye degradation was then predicted with the help of GC-MS analysis
technique.
Microbial Toxicity Testing of Dye Degraded Products
It becomes very important to know that whether the biodegradation of a harmful textile dye leads to
detoxification of the dye or not. Agar well bioassay is the most common technique used for the
evaluation of the microbial toxicity. This can be achieved by microbial toxicity tests with the original
dye and its biodegradation products. The microbial toxicity was tested on three test microorganisms
Viz. Azotobacter sp., Pseudomonas sp. Rhizobium sp. The 24 hours old culture of the test organisms
were used for the toxicity testing. After the confirmation of dye degradation, the degraded solution

(decolorized broth) obtained after treatment with the present organism was poured in the wells
prepared in nutrient agar previously spreaded with the test organisms. These plates were incubated at
ambient temperatures for 24 hours. The zone of inhibition around the wells proved the toxicity of the
dye and their degraded products.
Results
Isolation and Identification of organism
The promising bacterium was isolated from the acclimatized soil on nutrient agar containing 5.0%
NaCl and was identified by using biochemical observations and 16s rRNA analysis technique. From
the analysis the isolate was identified as Marinobacter salsuginis SAY-3. Biochemical results are given
in Table 1. The phylogenetic tree was developed by using Neighbor joining method by Kimura-2-
parameter with 1000 replicates in MEGA 4.0.10 (Figure 2).
99 DQ325514 Marinobacter koreensi

46 EU496088 Marinobacter santorin
56
EU047505 Marinobacter lacisals
54
DQ458821 Marinobacter pelagius
66
EF157832 Marinobacter segnicre
15 DQ414419 Marinobacter gudaonen
AY517632 Marinobacter flavimar
44 EF486354 Marinobacter salicamp

52 AJ609271 Marinobacter bryozoor
EU293413 Marinobacter zhejiang
49 SAY-3
93 EF028328 Marinobacter salsugin
EF660754 Marinobacter goseonge
AM503093 Marinobacter guineae
42
AY147906 Marinobacter lipolyti
35
DQ235263 Marinobacter vinifirm
50
89 AF479689 Marinobacter litorali
89 AY180101 Marinobacter excellen
0.005
Figure 2- Phylogenic tree of Marinobacter salsuginis SAY-3 (EMBL Accession No. HF545597).
Phylogenetic analysis of 16s rRNA gene sequence of Marinobacter salsuginis SAY-3. The percent
numbers at the nodes indicate the levels of bootstrap support based on neighbor-joining analyses of
1,000 replicates. The scale bar (0.005) indicates the genetic distance.

Table 1 Biochemicals of the isolate

Characteristics of isolate SAY-3 Marinobacter salsuginis SAY-3
Gram nature and Motility Gram negative motile rods
Optimal growth temperature (0C) 300C
Optimal NaCl Concentration (%) 5.0%
Utilization of -
D-Glucose +
Sucrose -
Mannitol +
Fructose +
Lactose -
Hydrolysis of -
Starch +
Gelatin -
Enzyme activity -
Amylase +
Oxidase +
Catalase +
Urease -
Nitrate reduction +
Optimization of Conditions
Effect of Static and Shaking Condition
The promising isolate was evaluated for its ability to decolorize the dye Sunset Yellow FCF in nutrient
broth medium containing (0.5%-18%) NaCl concentrations and 2000µg/ml concentration of dye. The
decolourization of Sunset Yellow FCF in static and shaking condition is given in Figure 3. The results
showed that static condition favored the maximum decolourization and degradation of the dye when
compared to shaking condition.
100
90
80
% Decolorization
70
60
50
40
30
20
Series1 Shaking Static
50 97
Figure 3 Percent Decolorization Sunset Yellow FCF in static and shaking condition in 24 hrs at max-
485nm
Effect of Temperature
The promising isolate showed best decolourization at 300C. It was found that the rate of
decolourization increased from 250C, 300C, 370C, 400C and 500C but it has been significantly affected
when temperature increased above the 400C (Figure 4). In the present study, the promising isolate
showed significant decolourization of Sunset Yellow FCF at 300C and increase in temperature above
370C decreases the decolourization activity of the strain.
95 95
85 85
% Decolorization
75 75
65 65
55 55
45 45
pH-5 pH-6 pH-7 pH-8 pH-9 pH-11
pH 50 80 90 97 80 45
Temp. 55 97 89 75 60
Figure 4 Percent Decolorization of Sunset Yellow FCF at different pH and temperatures in 24

hrs at max-485nm
Effect of pH
The effect of pH on the decolorization of the dye was also studied by inoculating the promising
organisms in a medium containing (0.5%-18%) NaCl concentrations and dye (2000µg/ml) at different
pH (5.0, 6.0, 7.0, 8.0, 9.0 and 11.0) and decolourization were measured at 485nm. The optimum pH for
maximum dye decolourization was found to be 8.0 (Figure 4). It was observed that the decolourization
of Sunset Yellow FCF increased with increase in pH but has been drastically affected after pH 8.0.
Percent Decolorization in presence of different 1% carbon and nitrogen sources
The selected bacterial isolate was further used for the decolorization assay in nutrient medium
containing (0.5%-15%) NaCl concentrations and 1% Glucose, 1% Starch and 1% Yeast extract as
carbon and nitrogen sources and 2000 g/ml concentration of dye. The results of percent decolorization
of dye Sunset Yellow FCF in presence of different carbon and nitrogen sources is given in Figure 5.

100
99.5
99
% Decolorization
98.5
98
97.5
97
NM 1% G 1%S 1%Y
Series1 97 98 98.55 100
Figure 5 Percent Decolorization in presence of different carbon and nitrogen sources (1%)
in 24 hrs at max- 485nm
Decolorization Assay
Percent Decolorization in Nutrient Broth
The promising bacterial isolate was used for to examine the decolorization in nutrient broth having
5.0% NaCl concentrations and 2000µg/ml concentration of dye. The results of percent decolorization
of dye Sunset Yellow FCF by the selected isolate is given in Figure 6.
Percent Decolorization in Half (½) Strength Nutrient Broth
The promising bacterial isolate was also examined for the decolorization in half strength nutrient broth
a having 5.0% NaCl concentrations and 2000µg/ml concentration of dye. The results of percent
decolorization of dye Sunset Yellow FCF in half strength nutrient broth is given in Figure 6.
Percent Decolorization by Cell-Free Extract
Further going, the promising isolate was examined for the decolorization in Cell-Free extract. The
results of percent decolorization of dye Sunset Yellow FCF in cell-free extract is given in Figure 6.
Percent COD reduction
After decolorization of the dye by the promising bacterial isolate, the percent COD reduction was
calculated and given in Figure 6.

98
96
% Decolorization
94
92
90
88
86
Series1 90 91 96 97
%COD R CFE HSNM NM
Figure 6 Percent Decolorization of Sunset Yellow FCF in Nutrient Broth, half Strength Nutrient Broth,
by Cell-Free extract and percent COD reduction in 24 hrs at max-485nm
Legend : %COD R= % COD Reduction, CFE = Cell Free Extract, HSNM = Half Strength Nutrient
Medium, NM = Nutrient Medium
Confirmation of Dye Degradation by GC-MS Technique
The GCMS analysis report showed that the dye Sunset Yellow FCF was degraded into smaller
fragments having molecular weight and are given in Figure 7 and Table 2 by Marinobacter salsuginis
SAY-3.

Figure 7 GCMS analysis report of degraded products of Sunset Yellow FCF dye by Marinobacter
salsuginis SAY-3
Detection of the Dye Degraded Products
The degraded products were detected by GC-MS technique and are given in Table 2.
Table 2- Detection of the Dye Degraded Products
Name of the Degraded Products Formed Molecular Weights of the Degraded
Products
sodium 3-(4-aminophenyl)trioxidane-1-thiolate 195.171509
sodium 5-amino-6-hydroxy-1,4-dihydronaphthalene-2- 263.245469
sulfonate
Aniline 93.12648
1-amino-5,8-dihydronaphthalen-2-ol 161.20044
5,8-dihydronaphthalen-1-amine 145.20104
Benzene 78.11184
(3Z)-penta-1,3-diene 68.11702
1,4-dihydronaphthalene 130.1864
(2Z)-but-2-ene 56.10632
(2Z,4E,7Z)-nona-2,4,7-triene 122.20746
(4E,6Z)-octa-1,4,6-triene 108.18088
Prediction of Dye Degradative Metabolic Pathway
The degradation products of Sunset Yellow FCF were analyzed by GC-MS and the results indicated
that the reductive cleavage (Figure 8) may be responsible for azo dye degradation by Marinobacter
salsuginis SAY-3.

HO
N N
O 3SNa
Sunset Yellow FCF SO 3Na

disodium
6-hydroxy-5-{[4-(sulfidotrioxidanyl)phenyl]di
azenyl}-1,4-dihydronaphthalene-2-sulfonate
Molecular Formula = C16H12N2Na2O7S2
Formula Weight = 454.385218
HO
NH2 H2N
O 3SNa
sodium 3-(4-aminophenyl)trioxidane-1-thiolate
Molecular Formula = C6H6NNaO3S sodium SO 3Na
Formula Weight = 195.171509 5-amino-6-hydroxy-1,4-dihydronaphthalene-2-sulfo
nate
NaSO 3 Molecular Formula = C10H10NNaO4S
NH2
HO SO 3Na
H2N
aniline
Molecular Formula = C6H7N
1-amino-5,8-dihydronaphthalen-2-ol
Molecular Formula = C10H11NO
benzene
Molecular Formula = C6H6 H2N
CH3
5,8-dihydronaphthalen-1-amine
CH2 Molecular Formula = C10H11N
(3Z)-penta-1,3-diene Formula Weight = 145.20104
Molecular Formula = C5H8
Formula Weight = 68.11702 NH3
CH3
CH3
(2Z)-but-2-ene 1,4-dihydronaphthalene
Molecular Formula = C4H8 Molecular Formula = C10H10
Formula Weight = 56.10632 Formula Weight = 130.1864
CH2 CH3
CH3 CH3
prop-1-ene
Formula Weight = 42.07974 (2Z,4E,7Z)-nona-2,4,7-triene
CH3
CH2
(4E,6Z)-octa-1,4,6-triene
CH2
CH2
(3E)-hepta-1,3,6-triene
H3C
CH2
(4E)-hexa-1,4-diene
Figure 8 - Probable Degradation Pathway of Sunset Yellow FCF by Marinobacter salsuginis

SAY-3
Microbial Toxicity Testing of Dye Degraded Products
Microbial toxicity of the textile azo dye Sunset Yellow FCF was studied on microorganisms Viz.
Azotobacter sp., Pseudomonas sp. and Rhizobium sp. The toxicity of the dye and its degradation

products was studied by the agar well assay. The results showed that the wells which were poured with
decolorized broth had no zone of inhibition and wells with original dye solution had a zone of
inhibition. This confirmed that the original dye solution 2000µg/ml was toxic to the bacteria but its
degradation products were non toxic to the bacteria.
Discussion
Al- Garni et al., (2013)11 studied the decolorization of Crystal violet by P. fluorescens and
Corynebacterium sp. Of which P. fluorescens was able to decolorize the concentrations of dye
150 g/ml after λ2 hours of incubation. The isolate decolorized the dye substrates and decolorizing
efficiency was dependent on the growth of the isolate in the tubes. In the present study the
acclimatized Marinobacter salsuginis SAY-3 showed on an average 97.00% decolorization of the azo
dye Sunset Yellow FCF. Chellaram et al., (2013)12 isolated 5 marine bacteria of which Pseudomonas
sp. MSB4 was tested for its degradation efficiency using 6 dyes Viz. Malachite green, Majenta MB,
Turquoise blue H, Orange Fast Boardeux RGB, Direct blue 2B and Direct Congo Red. Among the 5
strains, MSB4 showed maximum degradations activity against all the six dyes (Malachite green –
6mm, Majenta MB, Turquoise blue Ea, Orange fast boardeux – 4mm and Direct blue 2B, Direct
Congo Red – 2mm). The percentage of dye decolorization was observed against Majanta MB
(68.25%) followed by Orange Fast boardeux (64.10%), Congo red (63.55%), Malachite green
(61.89%) and Direct blue (22.09%), while lowest decolorization was observed against Turquoise blue
(12.06%) at 100ppm concentration. At 500ppm concentration the maximum percentage of dye
decolorization was observed against direct blue (9.03%). The presence of aromatic amine in the
microaerophilic stage and its absence in the aerobic stage indicated the presence of azoreductase
activity and an oxidative biodegradation process, respectively (Marimuthu, et al., 2013)13, however in
the present study, 91.00% decolorization was observed by cell free extract having 5.0% NaCl.
However, it has been reported that ligninolytic enzymes are better expressed in static cultures and
agitation of the cultures lead to decreased enzyme activity (Kaushik and Malik, 2009)14. In the present
study, maximum decolorization of Sunset Yellow FCF was observed under static conditions than in
agitated condition. In accordance with other reports, the present study revealed the excellent
decolorization (100%) capacity of Marinobacter salsuginis SAY-3 in presence of 1% yeast extract
when compared to starch and glucose which were used in this study but this decolorization was best
when compared with the nutrient medium. Gondaliya and Parikh (2012)15 reported the highest
percentage of decolorization 97.04% of Reactive Orange–16 was obtained by Serratia marcescens
when additional supplement of glucose (1g/l) was added in nutrient broth. These observations have
been similar with our results. Similar study was carried out by Krishnaswamy V. and Namasivayam V.

(2010)16 isolated a moderately halophilic bacterial consortium from saline environment which showed
maximum degradation of dye in the presence of yeast extract at 50g/L of NaCl and pH 7. Our results
were in strong agreement with the research conducted by Guo et al., (2008)17, in which the Halomonas
strains grew well and completely decolorized K-2BP where either yeast extract or peptone was present
in the medium; however, glucose, glycerol, sucrose, lactose and starch resulted in lower rates of
growth and decolorization of these dyes. Jagwani et al., (2013)18 reported maximum COD reduction of
67% and 83 % of ROM2R dye in 24 hours and 48 hours of inoculation respectively by Consortium
VSS. The mineralization of dye is indicated by this high rate of COD reduction (Hassan et al., 2002)19.
The present marine bacterium could reduce the COD up to 90.00% in 24 hours at 5.0% salinity.
Deshmukh et al., (2008)20 studied microbial toxicity of the undegraded and degraded dye on
agriculturally important microorganisms Azotobacter vinelandii and Pseudomonas aeruginosa. The
dye Navy blue Rx was found to be toxic whereas the dye metabolites were found to be stimulatory to
the microorganisms. Chen (2002)21 used the method in which the decolorized dye at the concentration
of 100mg/L was tested for its toxic effect on the agriculturally important soil bacterial flora Viz.
Bacillus cereus and Azotobacter sp. according to Mali et al.,(2000)22. No zone of inhibition was
observed in the treated dye, indicating that the biodegraded or decolorized products were non-toxic to
the tested beneficial bacterial flora of the soil Sidra Ilyas et al., (2012)23. In the present study, we also
have got the similar results. These results are in agreement with result obtained by Shertate and Thorat
(2013)24 who found that the metabolites produced after the biodegradation of Acid Orange 63 were non
toxic when compared with the original dye. Detoxification of the azo dyes by microorganisms
involved the conversion of azo-nitrogen to non-toxic metabolites (Chivukula and Renganathan,
(1995)25;Abadulla et al., (2000)26). In the present study, Marinobacter salsuginis SAY-3 degrade the
textile azo dye Sunset Yellow FCF at 5.0% NaCl into sodium 3-(4-aminophenyl)trioxidane-1-thiolate,
sodium 5-amino-6-hydroxy-1,4-dihydronaphthalene-2-sulfonate, aniline,1-amino-5,8-
dihydronaphthalen-2-ol, benzene, 5,8-dihydronaphthalen-1-amine, (2Z)-but-2-ene, 1,4-
dihydronaphthalene, prop-1-ene, (2Z,4E,7Z)-nona-2,4,7-triene, (4E,6Z)-octa-1,4,6-triene. Hebert and
Vreeland (1987)27 studied a halotolerant bacterium Halomonas halodurans, was capable of ortho
cleavage of aromatic compounds including benzoic acid. Sorensen et al., (2002)28 isolated a moderate
halophilic Marinobacter sp. able to completely degrade 1, 3-diphenylurea at 0.51M NaCl and 350C
and aniline was detected as an intermediate metabolite suggesting that a metabolic pathway involving
cleavage of the urea bridge between the phenyl structures. This study showed the Marinobacterium
capable of degrading the textile azo dye at high NaCl concentrations.

Conclusion
The study performed presently shown the major problem of the toxic textile dye Sunset Yellow FCF
employed in the study which is discharged from the textile industries located on the coastal area near
to the sea in the marine environment and reflecting the possible solution associated with the
degradation of the toxic dye under high saline condition. Therefore from the present study we can say
that for the treatment of the dye and dye containing marine wastewater, the biological method which
uses marine microorganisms can be useful, beneficial, inexpensive, eco-friendly and leading to a
Green Technology.
Acknowledgments: We would like to offer our sincere thanks to Principal, Shri Shivaji
Mahavidyalaya, Barshi for providing the Laboratory facilities, National Center for Cell Sciences
(NCCS), Pune for valuable support in the identification of the isolates and IIT, Powai, Mumbai for
their help in working out the samples with GC-MS analysis.
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Detoxification of A Harmful Textile Azo

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42 | P a g e International Standard Serial Number (ISSN): 2319-8141

INTERNATIONAL JOURNAL OF UNIVERSAL

DETOXIFICATION OF A HARMFUL TEXTILE AZO DYE SUNSET

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Figure 1 Structure and properties of the dye Sunset Yellow FCF

Acclimatization and isolation of microorganisms

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99 DQ325514 Marinobacter koreensi

44 EF486354 Marinobacter salicamp

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Table 1 Biochemicals of the isolate

Figure 4 Percent Decolorization of Sunset Yellow FCF at different pH and temperatures in 24

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Sunset Yellow FCF SO 3Na

Figure 8 - Probable Degradation Pathway of Sunset Yellow FCF by Marinobacter salsuginis

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8. ROBINSON, T., MCMULLAN, G., MARCHANT, R. 2001. Remediation of dyes in textile

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