Bioresource Technology 81 (2002) 155157

Short communication

Decolourisation of the textile dye Astrazon Red FBL

by Funalia trogii pellets
O. Yesilada *, S. Cing, D. Asma
Art and Science Faculty, Department of Biology, Inonu University, 44069 Malatya, Turkey
Received 11 April 2001; received in revised form 28 June 2001; accepted 4 July 2001

Abstract
The eects of various conditions such as initial pH, dye concentrations, amount of pellet, temperature and agitation on deco-
lourising activity of Funalia trogii were investigated. These, except initial pH, were all found to be important for dye decolourising
activity of F. trogii. The decolourisation of the dye involved adsorption of the dye compound by fungal pellets at the initial stage,
followed by the decolourisation through microbial metabolism. Heat-killed pellets were also tested for their ability to decolourise
Astrazon Red dye. These pellets adsorbed the dye and 55% decolourisation was obtained in 24 h. But at the second cycle there was
only 24% decolourisation. Our observation showed that Astrazon Red dye decolourisation by heat-killed pellets was mainly due to
biosorption. The longevity of the decolourisation activity of F. trogii pellets was also investigated in repeated batch mode. Variations
in the amount of pellet increased % decolourisation and stability of pellets. 2001 Elsevier Science Ltd. All rights reserved.

Keywords: Textile dye; Dye; White rot fungi; Funalia trogii; Decolourisation

1. Introduction (Kumar et al., 1998; Yesilada et al., 1999; Kapdan et al.,

Synthetic dyes are released into the environment in
industrial euents from textiles and the dyestu indus-
tries (Chang and Kuo, 2000). The removal of colour
from this type of wastewater is often more important
than the removal of the soluble colourless organic sub-
stances which usually contribute the major fraction of
the chemical and biochemical oxygen demand (Banat
et al., 1996). The elimination of coloured substances in
wastewater is based mainly on physical or chemical
methods. All of these methods have shortcomings. An
eective and inexpensive alternative would be of great
value (Zhang and Yu, 2000).
Textile dyes are also relatively resistant to microbial
degradation. Anaerobic microorganisms are able to
degrade some dyes. But aromatic amines produced by
these organisms may be toxic and carcinogenic (Meyer,
1981). Because conventional wastewater treatment sys-
tems generally do not remove all of the dyes, wastewa-
ters from industry and from dye manufacturers result in
pollution of the environment. White rot fungi can de-
grade a wide variety of structurally diverse pollutants
2000; Robles et al., 2000). In recent years many studies
have demonstrated that the white rot fungi were able to
decolourise dyes (Yesilada, 1995; Yesilada and Ozcan,
1998; Sani et al., 1998; Swamy and Ramsay, 1999). Most
researches on dye decolourisation involve two fungi
Phanerochaete chrysosporium and Coriolus versicolor.
Funalia trogii degrades and decolourises recalcitrant ef-
uents such as olive oil mill and alcohol factory waste-
water (Yesilada et al., 1998) and is therefore a good
candidate for treatment of dyes and textile wastewater.
There is almost no study on textile dye decolourising
ability of F. trogii and to our knowledge, Astrazon dyes,
that are widely used basic textile dyes, have not been
investigated for decolorisation by white rot fungi before.
In this paper we reported studies on the utilisation of
white rot fungus, F. trogii ATTC 200800, pellets for the
decolourisation of Astrazon Red FBL.
2. Methods
2.1. Pellet preparation

*
Corresponding author. Tel.: +90-42234-10010x3729; fax: +90-422-
F. trogii ATCC 200800 was cultured at 30 C on slant
3410037. SDA. After 1 week, a conidial suspension was prepared
E-mail address: oyesilada@inonu.edu.tr (O. Yesilada). and used for the cultivation of inoculum. Five ml of the

0960-8524/02/$ - see front matter 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 6 0 - 8 5 2 4 ( 0 1 ) 0 0 1 1 7 - 1
156 O. Yesilada et al. / Bioresource Technology 81 (2002) 155157

suspension was transferred into a 250 ml ask with 100
ml Sabouraud dextrose broth (SDB). It was incubated
for 4 days at 30 C in a shaking incubator (150 rpm).
After incubation, the culture was homogenised and used
for inoculation of fresh SDB (2 ml homogenate/100 ml
SDB). The fungal pellets were harvested after cultivation
and then used in the decolourisation experiments.
2.2. Decolourisation experiments
Synthetic dye test solutions were prepared by dis-
solving the dye in distilled water at concentrations of 0
1500 mg/l. Unless otherwise stated, the agitation rate
and temperature were 150 rpm and 30 C, respectively.
The eect of temperature, pH, dye concentration, agi-
tation and amount of pellet on decolourising ability of
F. trogii was studied. To test the eect of pH change on
dye decolourising, the buered test solutions without
fungi were used. The absorbance of this dye was not
aected over the range of pH 611 and showed that the
observed decolourisation was not due to a pH change.
Flasks which contained only dye and distilled water but
no fungi were used as controls.
2.3. Repeated batch experiments
A fresh test solution was rst inoculated with pellets.
After 24 h, the decolourised medium was discharged and
replaced with fresh test solution for the next cycle of
decolourisation by the same pellets.
2.4. Assay
Decolourisation was determined through measure-
ments of culture supernatant absorption at Astrazon
Red dye kmax (530 nm) using a spectrophotometer.
Percentage decolourisation was calculated from ab-
sorption values obtained against the controls. Dry
weight of pellets was obtained by ltering cultures
through lter paper and drying it to constant weight at
65 C. Results are the mean of at least three replicates.
3. Results and discussion
as the culture aged it was degraded from the mycelia.
The decolourising activity was slightly changed by agi-
tation. The highest decolourisation was obtained at 100
150 rpm. The optimum temperature for decolourisation
was found to be about 30 C. To nd the best initial pH
for decolourisation by F. trogii pellets, shake asks ex-
periments (150 rpm at 30 C) were conducted at dierent
initial pH values. The results showed that Astrazon Red
FBL was easily decolourised in all the initial pH values
used (pH 611). This gives the advantage that it is not
necessary to adjust the initial pH of this type of dye
euent.
3.2. Repeated batch experiments
The longevity of decolourisation activity of F. trogii
pellets was investigated in repeated batch decolourisa-
tion tests. As shown in Table 1, increase in amounts of
pellet positively aected the longevity of the decolouri-
sation activity. Colour removal eciency within 5 days
rapidly increased with the amounts of pellets used. On
the other hand, a decrease in the dye decolourisation
capability of pellets during the repeated batch mode
occurred with increasing dye concentration. Therefore,
high concentration of the dye showed a toxic eect that
adversely aected the performance. There are some re-
ports on the toxicity and genotoxicity of textile dyes and
wastewater (Walthall and Stark, 1999; Al-Sabti, 2000;
Sharma and Sobti, 2000). Hu and Wu (2001) determined
the toxic eect of textile dye RP2 B on Anabaena sp. and
reported that the removal of dyes or treatment of dyeing
wastewater is an urgent task for the sound management
Table 1
Decolourisation (%) of Astrazon Red dye by F. trogii using dierent
pellet amounts and dye concentrations
Number of times pellets used Dye concentration (mg/l)
13 66 132 264
Amount of pellet: 60 mg/50 ml
1 94 95 90 76
2 98 95 85 50
3 96 91 57 14
4 92 65 0 0
5 95 22 0 0

3.1. The eect of various conditions on decolourisation Amount of pellet: 150 mg/50 ml
1 93 96 96 95
2 98 99 98 93
Colour removal was observed to occur rapidly and
3 97 98 96 73
72% colour removal had occurred within 2 h. The 4 93 95 75 24
maximum decolourisation was achieved within 24 h. 5 95 75 70 0
Macroscopic and microscopic observations showed that
Amount of pellet: 370 mg/50 ml
the decolourisation process involved initial adsorption 1 95 98 97 97
of the dye compounds, which was followed by decolo- 2 98 99 99 99
urisation through microbial metabolism. Zheng et al. 3 98 99 98 97
(1999) reported the same observation that Poly R-478 4 96 98 94 70
5 94 91 69 28
was initially adsorbed onto the Penicillium mycelia and
 O. Yesilada et al. / Bioresource Technology 81 (2002) 155157
of the environment. At pellet amount of 60 mg/50 ml
decolourisation yield was initially good but declined
sharply on reuse of pellets especially at dye concentra-
tions above 13 mg/l. When the pellet amount of 370 mg/
50 ml and dye concentration of 132 mg/l were used, the
pellets retained their decolourisation activity for 5 days.
But when pellet amount of 370 mg/50 ml and dye con-
centration of 264 mg/l were used, the pellets retained
their decolourisation performance for 4 days. For dye
concentrations lower than 264 mg/l, the decolourisation
performances were stable for 5 days. More than 90%
reduction in colour intensity was achieved after the fth
cycle by using 13 or 66 mg/l of dye (for pellet concen-
trations of 150 and 370 mg/l).
These results showed that high pellet amounts gave
good decolourisation yield during the long-term opera-
tion. It was reported that dye concentration in dyehouse
euent is 1050 mg/l (Nigam et al., 2000). It can be
completely removed by F. trogii pellets and pellets can
be used several times. Yang and Yu (1996) reported that
it was possible to decolourise Red 533 by immobilised P.
chysosporium and this immobilised fungus could main-
tain high decolourising activity for a long-term opera-
tion. It was also stated that an increase in dye content
led to a decrement in decolourisation eciency. Simi-
larly, Knapp et al. (1997) and Zhang et al. (1999) re-
ported high and stable decolourising activity of F29
pellets during the repeated reuse.
Our results showed that pellets of this fungus could
eectively decolourise this basic dye and it was also
possible to induce the dye decolourising activity of this
fungus by carefully selecting the optimal conditions.
This biotechnological process has the advantage of ef-
fectively decolourising the basic dye Astrazon Red FBL
in distilled water without any other compound. Addi-
tionally, pellets could be used several times and still
maintain high decolourisation activity. Using fungal
pellets would allow treatment of euents with varying
compositions. It is possible to decolourise high concen-
tration of dyes, normally toxic at low concentrations, by
fungal pellets.
Acknowledgements
This research was supported by Turkish Republic
Prime Ministry State Planning Organisation (project no.
97K121480).
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