Decolorization of Indigosol Blue Dye Using Trametes versicolor F200 and Aspergillus sp

Diterima : 29/08/2019 Disetujui : 23/07/2020 Abstract The dyeing process of batik eventually produces much of wastewater. The difficult degradation and the dangers posed within the synthetic dyes are the main concerns in finding efficient wastewater treatment. Biological treatment has been known to be an effective technique of reducing or eliminating color intensity in wastewater. Fungi is one organism that can decompose many environmental pollutants. The aims of this research were to determine the ability of fungal isolates in decolorizing the synthetic dyes and analyzed which treatment has the highest decolorization percentage. Fungal isolates of Trametes versicolor F200 and Aspergillus sp. were used as a biological agent to decolorize of Indigosol Blue dye. The decolorization percentage was analyzed by spectrophotometer method. The result showed that T. versicolor F200 and Aspergillus sp. able to decolorize Indigosol Blue dye. The decolorization treatment of Indigosol Blue dyes using T. versicolor F200 showed the highest decolorization percentage reaching 97.21%. Keyword: Aspergillus sp., Decolorization, Synthetic dyes, Trametes versicolor F200


INTRODUCTION
Batik is a traditional art that has wisdom value and has been a part of Indonesian culture for a long time. Nowadays, batik industries are dominantly used the synthetic dye due to has various types of color, easier to be found, used in large scale production, and more simply practical use. Indigoid dye is one of the synthetic dyes that are often used in industrial batik.
The dyeing process eventually produces much wastewater, which contains a lot of organic compounds, smelly, and stained. The direct disposal of wastewater into the waters without any treatment will cause many environmental problems, such as generates lethal conditions for the aquatic organism and potentially affected to human health. Various methods of wastewater treatment to remove dye have been done, for example, chemical, physical and biological treatment. However, the physical and chemical treatments have a potential high cost and produce another toxic molecule (Namboodri et al., 1994;Slokar & Le Marechal, 1998;Tadda et al., 2016). Therefore, biological treatment more recommended and effective for decolorizing dyes because it takes place naturally, low cost, and there is no residual reaction that requires further processing (Sorta, 2013;Dewi et al., 2016).
Trametes versicolor and Aspergillus sp. have been known able to decolorize several of synthetic dyes. A recent study by Yang et al. (2017) showed that T. versicolor CBR43 able to decolorize of acid (Acid Red 114 and Acid Black 172), disperse (Disperse Black 1) and reactive dyes (Reactive Red 120, Reactive Blue 4, Reactive Orange 16, and Reactive Black 5). The decolorization of Acid Red 114 up to 86.5% over 2-4 days. Moreover, Acid Black 172 were decolorized attain 91.2% by six days. The Disperse Black 1 decolorized up to 80.2% in 9 days. Furthermore, the reduction of each reactive dyes reaches 96-97% dye removal at six days. Furthermore, the study by Hemalatha et al. (2016) reported that Aspergillus sp. collected from marine soil in Bay of Bengal Kakinada can decolorize several synthetic dyes. Aspergillus sp. showed 75% dye removal in Congo Red, 60% in Malachite Green, 45% in Brilliant Green, and 45% in Methylene Blue after five days incubation time. The change and disappearance of color contained in synthetic dyes indicate the degradation process carried out by fungal isolate.
The decolorization mechanism consists of adsorption, enzymatic degradation, and utilization as a carbon source. Adsorption is the primary step that occurs in the separation of dye molecular substance from the wastewater. Furthermore, the degradation process is the breakdown of complex molecules into simple compounds by ligninolytic enzymes (MnP, LiP, and laccase) produced by mycelial fungi (Setiadi, 2002). These enzymes have a role in degrading synthetic dye molecule through redox reaction, which completely oxidized become water, carbon dioxide, and or any other inorganic compound (Bennet & Fasion, 1994). Finally, the cleaved molecules of dye are utilized by fungi as a carbon source (Singh, 2006). This research aimed to determine the ability of T. versicolor F200 and Aspergillus sp. to decolorize of Indigosol Blue dyes and analyzed which treatment that has the highest decolorization percentage.

MATERIALS AND METHOD Chemical
Chemical compounds used in this research were obtained from The Research Center for Chemistry -Indonesian Institute of Science (LIPI) Puspiptek area, Serpong Tangerang. The Indigosol Blue dye was obtained from the batik industry in Banyumas, Central Java.

Fungal isolate
The fungal isolates used in this research are Trametes versicolor F200 obtained from Indonesia Culture Collection Research Center (InaCC) -LIPI, and Aspergillus sp. which obtained from the isolation process of Indigosol Blue effluent (Dewi et al., 2018).

Rejuvenation of Fungal Isolate
T. versicolor F200 and Aspergillus sp. were maintained in potato dextrose agar (PDA) medium in the Petri dish at 4°C until use. Fungal isolates are rejuvenated in PDA medium on the Petri dishes for seven days at room temperature.
Spectrophotometrically UV-vis analyzed the treatments and control at 0 hours and 24 hours. The change in absorbance value of Indigosol Blue was determined at 604.5 nm. The percentage of decolorization (D%) was calculated as follows: x 100 Detail: D% : Decolorization percentage A0 : The initial dye absorbance At : The absorbance at the t incubation period

RESULT AND DISCUSSION
The Trametes versicolor F200 and Aspergillus sp. were able to decolorize Indigosol Blue dyes. The average of decolorization percentage on Indigosol Blue dyes using T. versicolor F200 showed 97.21% dye removal whereas using Aspergillus sp. 60.63% and also control 1.15%. It is accordance with the study by Young & Yu (1997), T. versicolor able to decolorize indigo dye. 40-50 mg/L concentration of Indigosol Carmine dye was removed up to 92.8% at nine days incubation period. Furthermore, six different types of Aspergillus spp. in the study by Dewi et al. (2018) show decolorization activity against the batik dye effluent containing Indigosol Blue 04B. The highest decolorization percentages reach 99.05, 98.50, and 99.90%.

Trametes versicolor F200
Aspergillus sp. Control T. versicolor F200 showed higher ability than Aspergillus sp. in decolorizing of Indigosol Blue dyes. It is indicated that fungal strain affects the decolorization rate. The research by Park et al. (2004) reported that the decolorization rate of synthetic dyes was depended on fungal specificity towards synthetic dyes. Based on the previous description, it also indicates T. versicolor F200 from Basidiomycota has a greater ability than Aspergillus sp. (Ascomycota). According to the study by Claus et al. (2002), Basidiomycota has higher oxidative capacities and more efficient to decolorize of high concentration of synthetic dyes rather than Ascomycota. Decolorization rate by Polyporus pinistus and T.
versicolor from Basidiomycota were significantly higher than Myceliophthora thermophila (Ascomycota). At 0-100 ppm of dye concentration, P. pinisitus and T. versicolor are able decolorize indigoid Acid Blue 24 reach 65-80%, and anthraquinone Reactive Blue 19 up to 70-85% in 4 hours incubation period while M. thermophila exhibit slow decolorization rate and progressively decrease as long as the increasing of dye concentration.
The mechanism of decolorization can be occurred by non-enzymatically and enzymatically (Singh et al. 2006). Non-enzymatic mechanism occurs through the adsorption process, which is defined as the accumulation of toxins through the cells (Ngo et al. 2016). The color changes of fungal mycelium evidence this. The isolate of T. versicolor F200 showed the color change of mycelium from initial white to bluish. Similarly, Aspergillus sp. was turned into dark. It is in accordance with the statement of Yulisna (2000), the color changes of fungal mycelium indicates the decolorization process caused by the mechanism of dye adsorption by fungi. The color changes of T. versicolor F200 and Aspergillus sp. can be seen in  The enzymatic mechanism involves the secretion of the ligninolytic enzyme. Ligninolytic enzymes are an extracellular oxidative enzyme which responsible for the degradation of the aromatics compound. Ligninolytic enzymes consist of Manganese Peroxidase (MnP), Lignin Peroxidase (LiP) and laccase (Mester & Tien, 2000). These enzymes can degrade substrate nonspecifically. Therefore, fungi can decolorize various synthetic dyes. Ligninolytic enzymes are responsible for the breakdown of aromatic compounds into water, CO2, and other inorganic product (Hammel, 1997;Bennett & Faison, 1997).
The decolorization activity of T. versicolor F200 towards Indigosol Blue dye showed the highest decolorization percentage, reach 97.21% dye removal. Benito et al. (1997), T. versicolor can decolorize of synthetic dyes. As much as 10% of total color removed by adsorption and 90% removed by enzymatically of fungal mycellium. Study by Floudas et al. (2012), T. versicolor genetically showed the presence of 26 class II peroxidases (peroxidase) and ten multicopper oxidase (laccase) genes. It is indicated that T. versicolor able to produce MnP, LiP, and laccase.

CONCLUSION
Trametes versicolor F200 and Aspergillus sp. can decolorize of Indigosol Blue dye. The decolorization treatment of Indigosol Blue dyes using T. versicolor F200 show the highest decolorization percentage, reach 97.21%.