Professional Documents
Culture Documents
a,b,*
Doutorado em Ci^
encias Biol
ogicas, Universidade Federal de Pernambuco, 50670-420 Recife-Pernambuco, Brazil
b
Departamento de Qumica e N
ucleo de Pesquisas em Ci^
encias Ambientais,
Universidade Cat
olica de Pernambuco, 50050-590 Recife-Pernambuco, Brazil
Received 9 September 2002; received in revised form 15 May 2003; accepted 16 May 2003
Abstract
The inappropriate disposal of dyes in wastewater constitutes an environmental problem and can cause damage to the ecosystem.
Alternative treatments have been reported that fungi are particularly eective in the decolorization of textile euents. The decolorization of dyes with dierent molecular structures by Cunninghamella elegans was evaluated under several media conditions. The
decolorization procedures consisted of adding 72 h of mycelium into the culture medium containing either orange or reactive black
or reactive red or a mixture of these dyes in the presence or absence of sucrose and/or peptone. The decolorization prole was highly
dependent upon the incubation time, the molecular structure of the dye and presence or absence of co-substrates. The presence of
sucrose or both sucrose and peptone signicantly increased the decolorization of the solutions, however, the presence of only the
nitrogen source suppressed it. The ultraviolet spectra of the solutions before and after decolorization suggested the occurrence of
biodegradation in addition to the biosorption of the dyes. All tested dyes, except for the reactive black, caused inhibition of respiration of Escherichia coli, which suggested that toxic metabolites were produced.
2003 Elsevier Ltd. All rights reserved.
Keywords: Zygomycetes; Biodegradation; Biosorption; Textile dyes; Toxicity
1. Introduction
The reactive azo dyes are characterized by the presence of a nitrogennitrogen double bond (N@N),
namely the azo group, which is bound to aromatic
groups. Usually during the textile processing, around
3070% of the amount of the dye used is hydrolyzed and
eliminated into the wastewater (Bumpus, 1995).
In order to meet the criteria necessary for industrial
applications, these dyes present a diversity of colors,
molecular structures and resistance to fading upon exposure to light, water and many chemical compounds
(Correia et al., 1994). These required criteria thus yield
compounds that cause serious environmental pollution
problems. As a result of the environmental legislation,
industries are being forced to treat dye contaminated
their euents (Robinson et al., 2001). Due to their genetic diversity and metabolic versatility, microorganisms
*
Corresponding author. Tel.: +55-81-32164017; fax: +55-8132714043.
E-mail address: takaki@unicap.br (G.M. Campos-Takaki).
0960-8524/$ - see front matter 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0960-8524(03)00153-6
have become a viable alternative to remediate the pollution problem caused by reactive azo dyes (Alexander,
1994). Currently, bioremediation is becoming important
because it is cost eective, environmentally friendly, and produces less sludge (Banat et al., 1996; Robinson et al., 2001).
Many bacteria are able to degrade azo reactive dyes
aerobically and anaerobically (Tan et al., 1999); however, in many cases the metabolic products, usually aromatic amines (Hu, 2001), are toxic or even more toxic
than the starting azo dyes. Phanerochaete chrysosporium
has been reported to eciently degrade azo reactive dyes
such as Orange II, Congo red e Tropaeolim (Cripps
et al., 1990). In addition, it has recently been demonstrated that peroxidases (LiP and MnP), phenoloxidases
(laccases), and dioxygenases can act on specic recalcitrant pollutants by precipitation or transforming them
into other products, thus allowing for a possible better
nal treatment (Duran and Esposito, 2000).
It has been shown that some species of Basidiomycetes such as Phlebia tremellosa (Kirby et al., 2000),
Irpex lacteus, Pleurotus ostreaus (Novotny et al., 2001)
and Trametes modesta (Nyanhongo et al., 2002), can
70
nambuco State, Brazil by Gomes et al. (2000). The isolation and identication of strain are according to
ODonnell (1979). The culture was maintained on Sabouraud dextrose agar at 4 C and deposited in Culture
Collection of Nucleus of Resource in Environmental
Sciences, Catholic University of Pernambuco, Recife,
PE, Brazil.
Selected features of the azo dyes studied are presented
in Table 1. The azo dyes C.I. reactive black-5 and C.I.
reactive red-198 were provided by the Suape T^extil Co.,
located in the City of Cabo, Pernambuco, Brazil, and
the orange II (C.I.15510) was obtained from Sigma
(Sigma-Aldrich Corporation, St. Louis, Missouri, USA).
Solutions of these dyes were prepared by dissolving a
given amount of the powder in distilled water and then
added to the medium in order to yield solutions with
nal concentrations of 0.025 mM for the pure dyes and
0.034 mM for the mixture of dyes.
2. Methods
2.1. Microorganism and dyes
C. elegans UCP 542 was isolated from mangrove
sediment collected in the City of Rio Formoso, Per-
Table 1
Structure of azo dyes and their respective wavelength maximum absorption (kmax )
Dyes
kmax (nm)
Orange II
485
Chemical structure
OH
NaO3S
Reactive black 5
597
OH NH2
NaO3 SOCH2 CH 2 O2 S
N N
NaO 3 S
SO3Na
517
H 3C
NH
N
SO3Na
N
OH
N
NH
N
N
Cl
NaO 3S
SO 3 Na
Medium
I
II
III
IV
X
X
X
X
X
X
X
X
X
X
X
71
structure, culture media, incubation period, and the interaction among them. The means of the signicantly
dierent main eects were compared by the Duncans
test at the 5% level using the Statistica program (Statsoft
Inc., 1997).
72
0.65
a
1.0
Medium I
0.9
0.55
0.8
0.50
0.6
0.5
b
a
0.3
ab
a
b
ab
bb
bcc
0.35
ab
0.30
bc
0.25
c
ab
b
0.05
a
b
ab
0.20
ab
b
c
0.10
0.0
0.40
0.15
ab
b
0.1
ab
ab
0.4
b b
Decolorization (%)
Decolorization (%)
0.45
0.7
0.2
0.60
0.00
12
24
72
48
(A)
96
12
168
120
24
72
48
(B)
Time (hours)
96
Time (hours)
Medium III
1.4
Medium IV
168
120
1.0
1.2
Decolorization (%)
Decolorization (%)
0.8
a
1.0
0.8
a
ab
0.6
b
0.4
ab
b
b
bb
b
a
0.4
bb
b
ab
bc
bb
0.0
c
b
0.0
12
(C)
0.2
aa
a
c
0.6
bc
a
0.2
24
48
72
96
120
168
Time (hours)
12
(D)
24
48
72
96
120
168
Time (hours)
Fig. 1. Eect of medium composition I decolorization of the dyes orange II (j), reactive black (), reactive red ( ) and mixture ( ) by C. elegans
542. The letters indicate average values which are statistically dierent at P 0:05, using the Duncan test.
70
60
Toxicity (%)
50
40
30
20
10
0
Orange II
Reactive black
Reactive red
Mixture
Dyes
Fig. 2. Toxicity of dyes to E. coli before (j) and after () treatment
with C. elegans UCP 542.
Medium II
Medium I
Medium III
1.0
Medium IV
0.8
Control
Absorbance
73
0.6
0.4
0.2
0.0
300
350
400
450
500
550
600
650
700
750
Wavelength (nm)
Fig. 3. UVVIS spectra of orange II azo dye before and after the
treatment with C. elegans UCP 542 on dierent media: control ( ),
medium I ( ), medium II (), medium III ( ) and medium IV ( ).
74
Absorbance
0.5
0.4
Control
0.3
Mixture after 24 h
0.2
Mixture after 96 h
0.1
0.0
300
350
400
450
500
550
600
650
700
750
Wavelength (nm)
dye, despite the fact that the studied dyes had a common
stilbene-azo structural motif. In addition, it has been
shown by toxicity tests and UVVIS spectral analysis
that the decolorization process involved biodegradation,
in addition to the visually observed biosorption.
Acknowledgements
The authors gratefully acknowledge Prof. Dr. Ricardo L. Longo (DQF/UFPE) for assistance on analytical interpretation of the experimental data and
review of the manuscript; Dra. Sandra G. Moraes
(IQM/UNICAMP, SP) for the toxicity tests and Dra.
Adriana M.Y. Melo (Pesquisadora CNPq/Embrapa
rido) for helping with the statistical analyses. We
Semi-A
would like to acknowledge the Brazilian agencies CNPq,
CAPES, the programs FINEP/CTPETRO/AVINA
GROUP and PRONEX for nancial support.
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