Bioremediation of xenobiotics use of dead fungal biomass as biosorbent

IJRET: International Journal of Research in Engineering and Technology eISSN: 2319-1163 | pISSN: 2321-7308
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Volume: 03 Issue: 01 | Jan-2014, Available @ http://www.ijret.org 565
BIOREMEDIATION OF XENOBIOTICS: USE OF DEAD FUNGAL
BIOMASS AS BIOSORBENT
Mahmooda Takey1
, Toufique Shaikh2
, Nitin Mane3
, D. R. Majumder4
1,2,3
Student, Department of Microbiology, Abeda Inamdar Senior College, 2390-KB Hidayatullah Road, Azam
Campus, Camp, Pune, India. 4
Head, Department of Microbiology, Abeda Inamdar Senior College, 2390-KB
Hidayatullah Road, Azam Campus, Camp, Pune-411001, India
Abstract
Biosorption is bioengineering where metabolism independent adsorption of xenobiotics to living or dead cells takes
place.Microorganisms dead or alive are successfully exploited for bioremediation of xenobiotics by biosorption. In the present
study bioremediation of xenobiotics of textile industry effluent was carried out by biosorption using dead fungus biomass of
Aspergillus flavus. The dead biomass of fungus Aspergillus flavus shows maximum biosorption for three toxic components of
textile industry effluent under different parameters. Methyl orange biosorption was found to be 53.62% at room temperature, at
pH 5.5, with biomass concentration of 2g/L having contact time of 40 min and the dye concentration was 1ppm. Chromium
biosorption was 72.18%, at pH 6, at room temperature with biomass concentration of 2g/L having contact time of 10 min and
solution concentration 200ppm. Lead biosorption was found to be 76.12%, at pH 7, at room temperature with biomass
concentration 2g/L having contact time of 40 min and solution concentration 1ppm. Desorption studies were also performed and
was found that dead fungal biomass can be reused further.
Key words: Bioengineering, Bioremediation, Biosorption, Textile industry effluent, Methyl Orange, Chromium, Lead,
Aspergillus flavus .
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1. INTRODUCTION
Textile industry is the second largest industry in India. It
makes use of natural and synthetic dyes, heavy metals,
mordants for manufacturing of textile fabrics. The
compositions of the effluent are heavy metals (chromium,
copper, lead, and arsenic), dyes (azo, reactive), surfactants,
etc [1, 2]. It is characterized by high values of COD, BOD,
suspended and dissolved solids [1]. Textile industry effluent
causes ground water pollution which is of major concern as
they exert harmful effect to environment and life forms by
causing carcinogenicity, skin irritation, multi-organ failure,
neurotoxicity etc. The conventional physical and chemical
methods are less effective, expensive and use of chemicals
leads to secondary pollution [3-5]. Activated carbon is
found to be effective for adsorption but its generation
requires high temperature which is not cost effective. An
alternative to all these treatment methods is use of
biological entities like microorganisms which can be used in
live or dead forms for biosorption .[ 3,6 and 7 ]
Azo dyes have been found to be mostly used in the textile
industries. They contain –N=N- group as chromophore
group [8 and 9].They are resistant to microbial and chemical
attack and are most recalcitrant compound which are
carcinogenic, mutagenic, triggers allergic reactions in
human. [1, and 10-15]. Heavy metal Cr is soluble in water,
carcinogenic, neurotoxic leading to brain and nerves
damage, multi-organ failure while Pb is known to interfere
with enzyme activities and formation of RBCs, anemia,
hepatitis, nephratic syndrome [16 and 17].
Adsorbates (xenobiotics) are solutes to be adsorbed while
Adsorbent (living or dead biomass) is the solid material or
support used for adsorption and hence it is not affected by
microbial physiology [18 and 19]. The dried dead microbial
biomass displays high affinity for metal ion [20].
In this study biosorption of azo dye Methyl Orange and
heavy metals such as Chromium and Lead have been studied
using dead biomass of fungal species Aspergillus flavus. Cr
is used as metallic mordant while lead (acetate salt) is
employed in textile imprinting [21]. For coloring of wools
and silk fibers azo dye MO is used [22 and 23]. Various
parameters for maximum sorption profile of these three
compounds were studied such as pH of solution, contact
time, concentration of solution, temperature, and biomass
concentration.
2. MATERIALS AND METHOD
2.1 Fungus isolation and biosorbent preparation
Fungus Aspergillus flavus (identified by ARI, Pune) was
isolated from soil. Aspergillus flavus was grown and
maintained on GPYA (Glucose Peptone Yeast Extract Agar)
medium [composition (g/L): glucose- 40; peptone- 5; yeast
extract- 5; agar- 30; pH-5.6] incubated at room temperature
for 48hrs. To obtain fungal biomass, Aspergillus flavus was
inoculated in same liquid medium and incubated at RT,
150rpm for 20 days. The biomass was autoclaved at 1210 C,
for 15min at 15lbs. Then it was filtered through Whattman
filter paper No.1, dried at 550 C for one week and then
crushed in mortar and pestle. This dried dead powdered

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biomass was used for biosorption study. SEM analysis of
dried fungi was studied.
2.2 Reagents Preparation and lambda max
determination
Three components methyl orange, chromium and lead were
used in this study. Lambda max of aqueous solution of
methyl orange and chromium was determined by using
spectrophotometer Visican 167 while for lead UV/Vis
spectrophotometer was used. The lambda max of methyl
orange, chromium and lead was found to be 460nm, 420nm
and 210nm respectively. Aqueous stock solution of 10ppm
for Pb and MO and 1000ppm for Cr were prepared in
distilled water.
2.3 Study of sorption profile
Sorption profile was studied by using 0.1g of biosorbent in
50ml of each solution in 100ml Erlenmeyer flask. The
contents were kept at static conditions, centrifuged at
4000rpm after 10min time interval to completely sediment
biosorbent, supernatant was used for estimation of
biosorption efficiency. Initial and final absorbance was
determined for each solution which was used for estimation
of biosorption. Biosorbent was dried and used for desorption
studies.
Effects of different parameters on sorption process were
studied including biosorbent concentration, initial
concentration of dye / heavy metal solution and pH of
solution. To observe initial concentration at which
maximum biosorption was achieved, concentration of MO
in range of 1 to 8 ppm, Pb in range of 1 to 5ppm and Cr
from 100 to 600ppm were studied. Similarly to see the
effects of pH various pH were studied for each of the
solution. Biosorbent concentration was investigated in the
range of 1 to 3g of biosorbent per liter of solution.
2.4 Calculation of % Biosorption by dead biomass
Biosorption efficiency of biosorbent was calculated by using
following formula:
% Biosorption = Initial Absorbance – Final Absorbance X100
Initial absorbance
2.5 Desorption and recycling of biosorbent (dead
biomass)
After determining the biosorption profile, the reuse and
recycling possibility of biosorbent was investigated. For
desorption of biosorbent 0.1M HCl was used for MO, Cr
and Pb. MO being soluble in water, 0.1M HCl was used for
desorption of heavy metals as well as MO.
3. RESULTS
3.1 SEM Analysis
SEM of dried dead Aspergillus flavus biomass was
performed to investigate the mesh size, shape and particle
size of the biosorbent (Fig. 1). It has been found that particle
size of dead Aspergillus flavus biosorbent varies between
1µm to 10µm.
(a) 1µm
(b) 5µm
© 10µm
Fig 1 - SEM images of dead Aspergillus flavus biosorbent
3.2 Effect of initial concentration
The effect of initial concentration of solution of MO, Cr and
Pb on biosorption was studied. It was seen that Aspergillus
flavus shows maximum biosorption for MO, Pb and Cr at
concentration of 1ppm, 1ppm and 200 ppm within 40min,
40min and 30min respectively (Fig. 2, 3, 4). Percent
biosorption was found to be 18.90% for MO, 52.57% for Pb
while for Cr 39.19%.

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Fig. 2: Biosorption of Pb by Aspergillus flavus
0
5
10
15
20
0 10 20 30 40
%Biosorption
Contact time (Min)
1 ppm
2 ppm
3 ppm
4 ppm
5 ppm
6 ppm
7 ppm
8 ppm
Fig. 3: Biosorption of Methyl Orange by Aspergillus flavus
Fig. 4: Biosorption of Cr by Aspergillus flavus
3.3 Effect of pH of solution
Maximum biosorption was recorded at pH 5.5, 6 and 7
within 40 ,10 and 40 for MO, Cr and Pb respectively. At pH
5.5 MO was adsorbed 53.62%, while at pH 6 Cr was found
to be 72.18% and 73.35% Pb was adsorbed (Fig. 5, 6 and 7).
It has been found that increase in pH leads to decrease in
biosorption of MO and other dyes.
Fig. 5: Effect of pH on biosorption of MO by Aspergillus
flavus
Fig. 6: Effect of pH on Biosorption of Cr by Aspergillus
flavus
Fig. 7: Effect of pH on Biosorption of Pb by Aspergillus
flavus
3.4 Effect of Biomass concentration
In our study we have found that 2g biosorbent shows
maximum biosorption for 1L of all three component
solutions. That is 2g/L biomass concentration is found to be
effective (Fig. 8, 9, 10).

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Fig. 8: Effect of biomass concentration on biosorption of
Methyl orange by Aspergillus flavus
Fig. 9: Effect of biomass concentration on biosorption of Cr
by Aspergillus flavus
Fig. 10: Effect of biomass on biosorption of Pb
byAspergillus flavus
3.5 Desorption experiments
In this study, desorption of MO, Cr and Pb were carried out
by using 0.1M HCl solution where complete desorption for
MO, Pb and Cr at 50 min, 30 min and 50 min were seen
respectively(Fig. 11)
Fig. 11: Desorption of MO, Cr and Pb from Aspergillus
flavus
4. DISCUSSION AND CONCLUSION
Increase in pH leads to decrease in biosorption of MO and
other dyes due to characteristics of biomass and MO [21].It
has been found that increase in biomass concentration
decreases the biosorption efficiency [16, 22]. This occurs
due to decrease in surface area on biosorbent for binding of
adsorbent [16]. Our findings are in sync with the previous
findings.
Humicola fuscoatra adsorbs methyl orange, though it shows
biosorption of 62.5% it requires a long contact time of 6 hrs
[26 and 28] whereas dead fungal biomass of Aspergillus
flavus adsorb 53.62 % MO with contact time of only 40
min.. Percentage of adsorption by Aspergillus niger of Cr
was found to be more by base treated as compared to acid
treated and untreated Cr solution [29].While untreated dead
fungal biomass of Aspergillus.flavus adsorb 72.18%, of Cr
with contact time of 10min. Aspergillus terreus have shown
biosorption of Pb upto 100mg/g FFB (Free Fungal biomass)
in 10 min contact time [30]. Our study shows biosorption of
76.12% of Pb which is comparable.
It has been found that 0.1M HCl is effective in desorption of
heavy metals [16]. The reuse of dead fungal biomass can be
done for 4-5 cycles of biosorption and desorption without
significant loss in biosorption capacity. Aspergillus terreus
is known to show biosorption potential 78% of Pb after five
continous cycles of desorption [30]. Aspergillus niger shows
biosorption of Cr upto 40% after five cycles of desorption
[29]. Comparitively in our study dead fungal biomass of
Aspergillus flavus shows biosorption capacity of Pb, Cr and
MO at 76%, 72% and 53% respectively after 5 cycles of
desorption processes. This dead fungal biomass of
Aspergillus flavus can be used for biosorption upto 5-6
times and after desorption it can be discarded into the soil to
increase the fertility of the soil and it is not harmful to soil
flora.
Biosorption capacity of the dead biosorbent is due to high
surface to volume ratio [24]. Being metabolic independent
process, there are no restrictions of enzymatic activities of
adsorption [25]. It is harmless, cheap, eco-friendly, highly

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efficient [26 and 27]. Recover and reuse of bioadsorbent is
possible [18]. Biomass generated at fermentation industries
can be used as biosorbent after killing it thereby making it a
best practice for GLP of fermentation industries [29]. Thus
better alternative for conventional methods of adsorption is
biosorption and it is necessary to make use of biosorption at
large scale for treatment of wastewater.
ACKNOWLEDGEMENT
We would like to thank Dr. E.M.Khan, Principal, Abeda
Inamdar Senior College, for provding us the necessary
infrastructure conducive for research.
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Immobilized in Natural Matrix. Polish Journal of
Microbiology, 2010, Vol. 59, No. 1, 37-44.
BIOGRAPHIES
Postgraduate student in Microbiology
Working as Associate Professor in the
Department of Microbiology, Abeda
Inamdar Senior College, Pune
Maharashtra, India since 1991.
Teaching Graduate and Post Graduate
classes. Actively involved in research in
two areas viz.
a) Biotransformation
b) Nanobiotechnology
c) Environmental Microbiology

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