Folia Microbiol. 47 (5), 639–642 (2003)
http://www.biomed.cas.cz/mbu/folia/
Decolorization of Textile Dyeing Wastewater by Phanerochaete chrysosporium S. CING, D. ASMA (HAMAMCI), E. APOHAN, O. YE
ILADA*
Department of Biology, Science and Art Faculty, nonu University, 44069 Malatya, Turkey e-mail
[email protected] Received 17 October 2002 Revised version 13 June 2003
ABSTRACT. The potential use of fungal pellets for decolorization of the textile dyeing wastewater was evaluated. The live pellets of the fungus Phanerochaete chrysosporium were found to remove more than 95 % of the color of this wastewater within 1 d. The dye-removal capacity was a function of time and was proportional to the agitation rate; the optimum temperature was 30 °C. Both live and dead pellets were further examined in a repeated-batch mode for 5 d. The decolorization performance of live pellets remained high and stable for 5 d and they showed twice to thrice higher decolorization capacity than dead pellets.
Worldwide, wastewaters from textile industries are discharged in large quantities into natural water bodies on a daily base (Meehan et al. 2000). Because conventional wastewater treatment systems generally do not remove all of the dyes, wastewaters from industry and from dye manufacturers give rise to the pollution of the environment. One of the possible alternatives for the degradation of this type of compounds is the use of white-rot fungi. These fungi can degrade a wide variety of structurally diverse pollutants (Bhatt et al. 2002). In recent years many studies have demonstrated that fungi were able to decolorize and remove (biosorption) dyes (Ye ilada 1995; Knapp et al. 1997; Sani et al. 1998; Tatarko and Bumpus 1998; Swamy and Ramsay 1999; Aksu and Tezer 2000; Novotný et al. 2001; Kahraman and Ye ilada 2001; Sam and Ye ilada 2001; Eichlerová et al. 2002; Rodríguez Couto et al. 2002; Ye ilada et al. 2002; Zilly et al. 2002). The indigo dye is extensively used by textile industries and is considered a recalcitrant substance which causes environmental concern (Balan and Monteiro 2001); this dye was shown to be decolorized by ligninolytic enzymes (Campos et al. 2001; Podgornik et al. 2001; Verma and Madamwar 2002) but no study has been published on wastewater-containing indigo-dye decolorization activity of pellets from white-rot fungi. In this paper, the effect of various a culture conditions (amount of pellets, temperature, agitation and incubation time) on textile dyeing wastewater decolorization activity of Phanerochaete chrysosporium pellets was tested and longevity of the decolorization activity of live and dead pellets under optimum conditions was investigated in a repeated-batch mode process. MATERIAL AND METHODS Indigo dye and wastewater. The indigo dye and textile dyeing wastewater were obtained from GAP Textile Co. (Turkey). The principal component of this wastewater was the indigo dye. This wastewater, taken from the textile stream, was poured into several 5-L plastic containers and kept at 4 qC. The composition of wastewater is Table I. Characteristics of textile dyeing shown in Table I. wastewater Fungus. The white-rot fungus Phanerochaete chrysosporium ME446 was maintained at 4 °C, after subculturing at 30 °C every Parameter Value 2–3 weeks on Sabouraud dextrose agar (SDA). Color, A665 1.26 Pellet preparation. The fungus was cultured at 30 °C on SDA COD, mg/L 2089 slants. After 1 week, spore suspension was prepared and used for Total solid, mg/L 13 the cultivation of inoculum. Five mL of the suspension was transSuspended solid, mg/L 6.45 ferred into 100 mL Sabouraud dextrose broth (SDB) in a 250-mL pH 11.1 a incubator (2.5 Hz) at flask. Incubations were done in a shaking *Corresponding author.
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30 °C for 4 d. After incubation, the culture was gently homogenized and used for the inoculation of fresh SDB. Fungal pellets were harvested by filtration from culture media and wet pellets were used in the decolorization for further experiments. Besides the use of these live pellets, dead pellets (autoclaved at 121 qC for 20 min) were also used for decolorization. Decolorization experiments. The pellets were used at three different concentrations (127, 255 and 462 mg per 50 mL) by inoculating into 50 mL wastewater in 250-mL flasks. Unless otherwise stated, the agitation rate and temperature were 2.5 Hz and 30 °C, respectively. The effect of temperature, agitation and the amount of live pellets on decolorization was studied. Also, the capacity of live pellets on decolorization was examined using only indigo dye in the water, given that this dye is the main component of wastewater used in our study. In order to test if there was any color change during the incubation period, a control experiment was carried out without pellets. Repeated-batch experiments. Live and dead pellets were used in a repeated-batch mode under optimum conditions five times with a residence time of 1 d. A fresh test solution was first inoculated with live or dead pellets (462 mg per 50mL). The decolorized medium was discharged and replaced with fresh test solution for the next cycle of decolorization by the same pellets after 1 d. Assays. The degree of decolorization was quantified by measuring changes of absorbance at its maximum wavelength (A ( 665) by using a Shimadzu 1601 UV/VIS spectrophotometer. The chemical oxygen demand (COD), total solid and suspended solid were determined according to Standard Methods (1979). Dry mass of pellets was obtained by filtering cultures through filter paper and drying them to constant mass at 65 °C. Results are the means rSD of at least three replicates. RESULTS AND DISCUSSION The main component of the wastewater used in this study was the indigo dye. We first studied the capacity of live P. chrysosporium pellets in color removal in indigo-containing water. Color removal percentages by the pellets determined within 1 d of incubation with dye concentrations 100, 200 and 400 mg/L were 96.5 r 0.1, 97.5 r 0.3 and 95 r 0.9 %. Balan and Monteiro (2001) reported a 75 % decolorization of indigo dye by P. chrysosporium within 4 d of incubation while Yang and You (1996) reported 30 % decolorization by the growing cells of P. chrysosporium after 9 d and suggested that ligninase was the major enzyme catalyzing dye decolorization. In further experiments, the live pellets showed a rapid wastewater decolorization activity. Color removal of 61 and 95 % was observed after 2-h and 1-d incubations, respectively (Table II). The pellets were Table II. Decolorization (%; after 2–24-h cultivation) of textile dyeing wastewater by live pellets Cultivation period, h %
2
4
6
8
24
61.1 r 3.2
63.5 r 0.6
65.3 r 1.8
68.5 r 1.3
94.9 r 3.1
dark blue after 2 h and pale blue after 1-d incubation. Intact pellets adsorbing dyes were deeply colored whereas those subject to degradation remained pale blue. This observation shows that decolorization by live pellets involves microbial metabolism. To test the effect of agitation on the decolorization process, live pellets were used under both static and agitated conditions. The highest decolorization of 95 % was obtained at an agitation frequency 2.5 Hz. A similar study by Knapp et al. (1997) showed that the decolorization of Orange II was a mere 45 % after a 23-h incubation in static conditions and 98 % in agitated conditions. In experiments done at various temperatures, the highest decolorization was obtained at 30 °C. Previous studies showed that the pH of the medium
Table III. Effect of the amount of live pellets, agitation and temperature on decolorization (%) of textile dyeing wastewater by live pellets Parameter
Value
Decolorization, %
Agitation frequency Hz
0a 0.8 1.6 2.5
48.9 r 2.1 54.6 r 2.1 85.8 r 6.3 94.9 r 3.1
Concentration of pellets g/L
2.54 5.10 9.24
85.5 r 1.8 95.3 r 2.0 94.9 r 3.1
Temperature qC
aStatic cultivation.
20 30 40
41.0 r 1.3 94.9 r 3.1 56.6 r 1.8
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may be adjusted for effective decolorization of textile wastewater and Orange II dye (Knapp et al. 1997; Shahvali et al. 2000). Here, we obtained high decolorization without adjusting the pH of the wastewater (Ye ilada et al. 2002). Controls containing wastewater but with no pellets showed no change in color. With dead pellets,the percentage of decolorization was substantially lower (33 % during 1 d). The effect of pellet concentration on decolorization was not pronounced in the studied concentration range (yields of 86, 95 and 95 % were determined for live pellets used at 2.54, 5.10 and 9.24 g/L, respectively; Table III). The longevity of decolorization activity of live and dead pellets was further investigated in repeated-batch decolorization mode (Table IV). The live pellets showed high and stable decolorization activity during a 5-d operation. Yang and Yu (1996) reported that immobilized P. chysosporium could maintain high Table IV. Decolorization (%) of textile dyeing wastewater with live and dead pellets in repeated-batch mode Number of runs with a pellet batch 1 2 3 4 5
Live pellets
Dead pellets
94.9 r 3.1 93.0 r 3.4 92.3 r 4.0 82.1 r 14.7 88.3 r 3.5
37.6 r 3.6 36.5 r 3.7 39.2 r 1.3 36.9 r 9.1 38.7 r 8.4
decolorization activity in a long-term operation. Similarly, Knapp et al. (1997) and Zhang et al. (1999) reported high and stable decolorizing activity of unidentified wood-rotting fungus F29 pellets during the repeated use. Recently, we reported that live pellets of the white-rot fungus Funalia trogii significantly decolorized Astrazone dye during a long-term cultivation (Ye ilada et al. 2002). In order to determine whether a nonsupplemented effluent could support cell growth we used gently homogenized live pellets and inoculated them into wastewater. Neither growth nor decolorization could be determined, probably due to an inhibitory effect of the effluent. Our results suggest that P. chrysosporium pellets can contribute to the decolorization and degradation of dyes in textile-industry effluents. This study was supported by a grant (no. 2000/16) from Research Fund Unit of nonu University (Turkey). REFERENCES AKSU Z., TEZER S.: Equilibrium and kinetic modeling of biosorption of Remazol Black B by Rhizopus arrhizus in a batch system: effect of temperature. Proc.Biochem. 36, 431–439 (2000). BALAN D.S.L., MONTEIRO R.T.R.: Docolorization of textile indigo dye by ligninolytic fungi. J Biotechnol. 89, 141–145 (2001). BHATT M., CAJTHAML T., ŠAŠEK V.: Mycoremediation of PAH-contaminated soil. Folia Microbiol. 47, 255–258 (2002). CAMPOS R., KANDELBAUER A., ROBRA K.H., CAVACO-PAULO A., GUBITZ G.M.: Indigo degradation with purified laccases from Trametes hirsuta and Sclerotium rolfsii. J.Biotechnol. 89, 131–139 (2001). EICHLEROVÁ I., HOMOLKA L., NERUD F.: Decolorization of synthetic dyes by Pleurotus ostreatus isolates differing in ligninolytic properties. Folia Microbiol. 47, 691–695 (2002). KAHRAMAN S., YE ILADA O.: Industrial and agricultural wastes as substrates for laccase production by white-rot fungi. Folia Microbiol. 46, 133–136 (2001). KNAPP J.S., ZHANG F.M., TAPLEY K.N.: Decolorization of Orange II by wood-rotting fungus. J.Chem.Technol.Biotechnol. 69, 289–296 (1997). MEEHAN C., BANAT I.M., MCMULLAN G., NIGAM P., AMYTH F., MARCHANT R.: Decolorization of Remazol Black B using a thermotolerant yeast, Kluyveromyces marxianus IMB3. Environ.Internat. 26, 75–79 (2000). NOVOTNÝ C., RAWAL B., BHATT M., PATEL M., ŠAŠEK V, MOLITORIS H.P.: Capacity of Irpex lacteus and Pleurotus ostreatus for decolorization of chemically different dyes. J.Biotechnol. 89, 113–122 (2001). PODGORNIK H., POLJANSEK I., PERDIH A.: Transformation of indigo carmine by Phanerochaete chrysosporium ligninolytic enzymes. Enzyme Microb.Technol. 29, 166–172 (2001). RODRÍGUEZ COUTO S., DOMÍNGUEZ A., SANROMÁN Á.: Production of manganese-dependent peroxidase in a new solid-state bioreactor by Phanerochaete chrysosporium grown on wood shavings. Application to the decolorization of synthetic dyes. Folia Microbiol. 47, 417–422 (2002). SAM M., YE ILADA O.: Decolorization of orange II dye by white rot fungi. Folia Microbiol. 46, 143–146 (2001). SANI R.K, AZMI W., BANERJEE U.C.: Comparison of static and shaken culture in the decolorization of textile dyes and dye effluents by Phanerochaete chrysosporium. Folia Microbiol. 43, 85–88 (1998). SHAHVALI M., ASSADI M.M., ROSTAMI K.: Effect of environmental parameters on decolorization of textile wastewater using Phanerochaete chrysosporium. Bioproc.Engineer. 23, 721–726 (2000). Standard Methods for the Examination of Water and Wastewater, 14th ed., p. 1138. American Public Health Association, Washington (DC) 1979.
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