Metabolite Identification from Biodegradation of Congo Red by Pichia sp.
Abstract
Azo dyes are commonly used in textile and paper industries. However, its improper disposal often results in polluting water bodies. Azo dyes can cause adverse health effects because of its carcinogenic properties. Various methods to remove azo dyes from water have been proposed, including biological methods such as biosorption and biodegradation. Biosorption and biodegradation were done by using bacteria, yeast or mold. In general, yeasts have some advantages for azo dyes degradation due to its faster growth compared to mold and better resistance against unfavorable environment compared to bacteria. Previously, we observed that yeast Pichia sp. have the ability to degrade Congo red, an azo dye. However, information regarding biodegradation of azo dyes by Pichia sp. are still limited. Therefore, in this study, we showed degradation of Congo red by Pichia sp. crude enzyme extract obtained from separating Pichia cells from medium by centrifugation, followed by identification of its biodegradation products. Biodegradation product was separated from enzyme by ethyl acetate and then Gas Chromatography-Mass Spectroscopy (GC-MS) method was employed to identify biodegradation product. Chromatogram results of GC-MS showed that Congo red were degraded into various products such as biphenyl, naphthalene and smaller molecules with 94 m/z and 51 m/z. These results suggest involvement of azo reductase and laccase-like enzymes which cleaves azo bonds and oxidize the dye molecules to smaller molecules. This study implies the use of Pichia sp. as a bioremediation agent for the removal of azo dyes.
Keywords: Biodegradation, Congo red, Pichia sp., metabolite identification, GC-MS
References
[1] Benkhaya, S., El Harfi, S., and El Harfi, A. “Classifications, Properties and Applications of Textile Dyes: A Review.” Applied Journal of Environmental Engineering Science, Vol. 3, No. 3, 2017, pp. 311–320.
[2] Saroj, S., Kumar, K., Pareek, N., Prasad, R., and Singh, R. P. “Biodegradation of Azo Dyes Acid Red 183, Direct Blue 15 and Direct Red 75 by the Isolate Penicillium Oxalicum SAR-3.” Chemosphere, Vol. 107, 2014, pp. 240–248.
[3] Lade, H., Govindwar, S., and Paul, D. “Mineralization and Detoxification of the Carcinogenic Azo Dye Congo Red and Real Textile Effluent by a Polyurethane Foam Immobilized Microbial Consortium in an Upflow Column Bioreactor.” International Journal of Environmental Research and Public Health, Vol. 12, No. 6, 2015, pp. 6894–6918. doi:10.3390/ijerph120606894.
[4] Shah, K. “Biodegradation of Azo Dye Compounds.” International Research Journal of Biochemistry and Biotechnology, Vol. 1, No. 2, 2014, pp. 5–13.
[5] Hassaan, M. A., and Nemr, A. E. “Health and Environmental Impacts of Dyes: Mini Review.” American Journal of Environmental Science and Engineering, Vol. 1, No. 3, 2017, pp. 64–67.
[6] Garg, S. K., and Tripathi, M. “Microbial Strategies for Discoloration and Detoxification of Azo Dyes from Textile Effluents.” Research Journal of Microbiology, Vol. 12, No. 1, 2017, pp. 1–19. doi:10.3923/jm.2017.1.19.
[7] Kaushik, P., and Malik, A. “Fungal Dye Decolourization: Recent Advances and Future Potential.” Environment International, Vol. 35, No. 1, 2009, pp. 127–141. doi:10.1016/j.envint.2008.05.010.
[8] Srinivasan, A., and Viraraghavan, T. “Decolorization of Dye Wastewaters by Biosorbents: A Review.” Journal of Environmental Management, Vol. 91, No. 10, 2010, pp. 1915–1929. doi:10.1016/j.jenvman.2010.05.003.
[9] Singh, R. L., Singh, P. K., and Singh, R. P. “Enzymatic Decolorization and Degradation of Azo Dyes – A Review.” International Biodeterioration & Biodegradation, Vol. 104, 2015, pp. 21–31. doi:10.1016/j.ibiod.2015.04.027.
[10] Olukanni, O. D. “Decolorization of Dyehouse Effluent and Biodegradation of Congo Red by Bacillus Thuringiensis RUN1.” Journal of Microbiology and Biotechnology, Vol. 23, No. 6, 2013, pp. 843–849. doi:10.4014/jmb.1211.11077.
[11] Mukherjee, T., and Das, M. “Degradation of Malachite Green by Enterobacter Asburiae Strain XJUHX- 4TM.” CLEAN – Soil, Air, Water, Vol. 42, No. 6, 2014, pp. 849–856. doi:10.1002/clen.201200246.
[12] Ilyas, S., Bukhari, D. A., and Rehman, A. “Decolorization of Synozol Red 6HBN by Yeast, Candida Tropicalis 4S, Isolated from Industrial Wastewater.” Pakistan Journal of Zoology, Vol. 47, No. 4, 2015, pp. 1181–1185.
[13] Acikgoz, C., Gül, Ü. D., Özan, K., and Borazan, A. A. “Degradation of Reactive Blue by the Mixed Culture of Aspergillus Versicolor and Rhizopus Arrhizus in Membrane Bioreactor (MBR) System.” Desalination and Water Treatment, Vol. 57, No. 8, 2016, pp. 3750–3756. doi:10.1080/19443994.2014.987173.
[14] Das, R., and De, A. “Decolorization Of Selective Textile Dyes Using Waterborne Pathogenic Bacterial Strains.” 2016. doi:10.5281/zenodo.225190.
[15] Laulund, S., Wind, A., Derkx, P., and Zuliani, V. “Regulatory and Safety Requirements for Food Cultures.”VMicroorganisms, Vol. 5, No. 2, 2017, pp. 28–41. doi:10.3390/microorganisms5020028.
[16] Tamang, J. P., Watanabe, K., and Holzapfel, W. H. “Review: Diversity of Microorganisms in Global Fer- mented Foods and Beverages.” Frontiers in Microbiology, Vol. 7, 2016. doi:10.3389/fmicb.2016.00377.
[17] Tati Barus, and Steffysia. “Genetic Diversity of Yeasts from Ragi Tape ‘Starter for Cassava and Glutinous Rice Fermentation from Indonesia’ Internal Transcribed Spacer (ITS) Region.” Merit Research Journal of Food Science and Technology, Vol. 1, No. 3, 2013, pp. 31–35.
[18] Qu, Y., Cao, X., Ma, Q., Shi, S., Tan, L., Li, X., Zhou, H., Zhang, X., and Zhou, J. “Aerobic Decolorization and Degradation of Acid Red B by a Newly Isolated Pichia Sp. TCL.” Journal of Hazardous Materials, Vol. 223–224, 2012, pp. 31–38. doi:10.1016/j.jhazmat.2012.04.034.
[19] Feng, C., Fang-yan, C., and Yu-bin, T. “Isolation, Identification of a Halotolerant Acid Red B Degrading Strain and Its Decolorization Performance.” APCBEE Procedia, Vol. 9, 2014, pp. 131–139. doi:10.1016/j.apcbee.2014.01.024.
[20] Rosu, C. M., Avadanei, M., Gherghel, D., Mihasan, M., Mihai, C., Trifan, A., Miron, A., and Vochita, G. “Biodegradation and Detoxification Efficiency of Azo-Dye Reactive Orange 16 by Pichia Kudriavzevii CR-Y103.” Water, Air, & Soil Pollution, Vol. 229, No. 1, 2018, p. 15. doi:10.1007/s11270-017-3668-y.
[21] Roșu, C. M., Vochița, G., Mihășan, M., Avădanei, M., Mihai, C. T., and Gherghel, D. “Performances of Pichia Kudriavzevii in Decolorization, Biodegradation, and Detoxification of C.I. Basic Blue 41 under Optimized Cultural Conditions.” Environmental Science and Pollution Research, Vol. 26, No. 1, 2019, pp. 431–445. doi:10.1007/s11356-018-3651-1.
[22] Sladewski, T. E., Shafer, A. M., and Hoag, C. M. “The Effect of Ionic Strength on the UV–Vis Spectrum of Congo Red in Aqueous Solution.” Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, Vol. 65, No. 3–4, 2006, pp. 985–987. doi:10.1016/j.saa.2006.02.003.
[23] Iwunze, M. O. “Aqueous Photophysical Parameters of Congo Red.” Spectroscopy Letters, Vol. 43, No. 1, 2010, pp. 16–21. doi:10.1080/00387010903278226.
[24] Ng, I.-S., Chen, T., Lin, R., Zhang, X., Ni, C., and Sun, D. “Decolorization of Textile Azo Dye and Congo Red by an Isolated Strain of the Dissimilatory Manganese-Reducing Bacterium Shewanella Xiamenensis BC01.” Applied Microbiology and Biotechnology, Vol. 98, No. 5, 2014, pp. 2297–2308. doi:10.1007/s00253-013-5151-z.