Experimental Design Optimization and Decolorization of an Azo Dye by Cross-Linked Peroxidase Aggregates

The immobilization of horseradish peroxidase in the form of cross-linked enzyme aggregates (HRP–CLEAs) was optimized by three parameters full factorial experimental design at two levels and the degradation of an azo dye was tested. The optimal immobilization conditions were estimated as 0.06 mg enzyme/mL (0.96 U), 3 % (v/v, Glutaraldehyde)) cross-linker ratio, and 6 mg /mL Bovine Serume Albumine amount, respectively. The effects of variables were analysed by responce surface plots and the enzyme concentration effect was insignificant for the tested levels. The maximum CLEAs activity was 0.2188 U and the kinetic parameters (Km and Vmax) were 0.0314 mM and 0.1044 mM/min while the calculated values for soluble enzyme were 0.06 mM and 0.468 mM/min, respectively. Hydrogen peroxide has inhibitory effect on enzyme aggregate activities at higher amounts than 0.012 mM. Under optimal conditions, a textile azo dye, Reactive Blue 160 was completely oxidized within 90 h and higher temperatures also showed promising results in dye degradation.  

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  • 1. Da Silva M. R., de Sà, L. R. V., Russo C., Scio E. Ferreira-Leitao V. S., Volume 2010, The Use of HRP in Decolorization of Reactive Dyes and Toxicological Evaluation of Their Products. SAGE-Hindawi Access to Research Enzyme Research, Article ID 703824, 7 pages http://dx.doi.org/10.4061/2010/703824.2. Levin L., Forchiassin F., Viale A., 2005, Ligninolytic enzyme production and dye decolorization by Trametes trogii: application of the Plackett–Burman experimental design to evaluate nutritional requirements. Process Biochemistry, 40, pp: 1381–1387. https://doi.org/10.1002/1521-3846(200105)21:2<179::AID-ABIO179>3.0.CO;2-2.3. Shaheen, R., Asghera M., Hussaina F., Bhatti H. N., 2017, Immobilized lignin peroxidase from Ganoderma lucidum IBL-05 with improved dye decolorization and cytotoxicity reduction properties. International Journal of Biological Macromolecules 103, pp: 57–64. https://doi.org/10.1016/j.ijbiomac.2017.04.040. 4. Robinson T., McMullan G., Marchant R. Nigam P., 2001, Remediation of dyes in textile effluent: a critical review on current treatment technologies with a proposed alternative. Bioresource Technology, 77, pp: 247-255. https://doi.org/10.1016/S0960-8524(00)00080-8.5. Rathnamsamy S. Singh R., Auxilia R., Vedhahari B.N., 2014, Extraction of peroxidasese from various plant sources and its biodegradation studies on phenolic compounds. BioTechnology, 9(4), pp: 160-165. 6. Pandey V. P., Awasthi M., Singh, S. Tiwari S., Dwivedi U. N., 2017, A Comprehensive Review on Function and Application of Plant Peroxidases. Biochem Anal Biochem 6, pp: 1-16. doi:10.4172/2161-1009.10003087. Kawakita H., 2012, Metal Recovery Using Polyphenols Prepared by Enzymatic Reactions of Horseradish Peroxidase. Science and Technology, 2 (1), pp:25-29 DOI: 10.5923/j.scit.20120201.05 8. Kariminiaae-Hamedaani H-R. Sakurai A. Sakakibara M., 2007, Decolorization of synthetic dyes by a new manganese peroxidase producing white rot fungus. Dyes Pigments, 72, pp: 157-162. doi:10.1016/j.dyepig.2005.08.010. 9. Hamad I. S. Ahmed A., Azim A., 2013, Biodegradation of phenols in wastewater using crude peroxidases five weed plants. J Chem & Pharmac R., 5 (4), pp: 60. 10. Bansal P., Dhir A., Prakash N. T., Sud D., July 2011, Environmental remediation of wastewater containing azo dyes with a heterostructured nanophotocatalyst. Indian Journal of Chemistry Vol. 50A, pp. 991-995. 11. Farias S., de Oliveira D., Ulson de Souza, A. A., Guelli S. M. A., de Souza U., Morgado A. F., 2017, Removal of reactıve blue 21 and Reactıve red 195 dyes usıng horseradısh Peroxıdase as catalyst Brazilian Journal of Chemical Engineering, July–September, Vol. 34(03), pp: 701–707. dx.doi.org/10.1590/0104-6632.20170343s20160091.12. Yinca Z., Yan L., Xueyong, G., Qiao W., and Xiaoping X., 2017, Decolorization of Color Index Acid Orange 20 buffer solution using horseradish peroxidase immobilized on modified PAN-beads. RSC Adv., 7, 18976–18986. DOI: 10.1039/c7ra01698k. 13. Grateron C., Barbosa O., Ruedaa N., Ortiz-L´opez C., Torres R., 2007, Azo dye decolorization by optimized cross linked enzyme aggregates (CLEAs) of a royal palm (Roystonea regia) peroxidase. Abstracts / Journal of Biotechnology 131S, pp: S74–S97.14. Sekuljica N. Z., Prlainovic N. Z., Jakovetic S. M., Grbavcic S. Z., Ognjanovi N. D., Knezevi-Jugovic Z. D., Mijin D. Z., 2016, Removal of Anthraquinone Dye by Cross-Linked Enzyme Aggregates From Fresh Horseradish Extract. Clean – Soil, Air, Water 44 (7), pp: 891–900. DOI: 10.1002/clen.201500766.15. Topçular C., Ayhan H., 2008, Carrier free cross-linked peroxidase aggregates Synthesis and Characterization. Hacettepe Journal of Biology & Chemistry,36(3), pp: 255-261.16. Ayhan F., Doğaç Y., Ayhan H., 2011, Cross-linked Glucose oxidase aggregates: Synthesis and Characterization. Hacettepe Journal of Biology & Chemistry, 39 (3), pp: 241-251.17. Claiborne A., Fridovich I., 1979, Chemical and Enzymatic Intermediates in the Peroxidation of o-Dianisidine by Horseradish Peroxidase. 1. Spectral Properties of the Products of Dianisidine Oxidation. Biochemistry, 18(11), pp: 2325. 18. Bania I., Mahanta R., May 2012, Evaluatıon of peroxıdases from varıous plant sources. International Journal of Scientific and Research Publications, Volume 2(5), pp: 1-5. 19. Montgomery D. C., George C. R., 2003, Applied Statistics and Probability for Engineers, 3rd edition John Wiley & Sons, Singapore.20. Engineering Statistics Handbook, Improve-Process Improvement, Full Factorial Designs, https://www.itl.nist.gov/div898/handbook/pri/section3/pri3332.htm, Date created: 6/01/2003, Last updated: 10/30/2013, (Accessed 06/08/2018) 21. Bailey J.E., Ollis D.F., Biochemical Engineering Fundamentals. McGraw-Hill Book Company, Singapore, 1986.22. Mohan S. V., Prasad K. K., Rao N. C., Sarma P.N., 2005, Acid azo dye degradation by free and immobilized horseradish peroxidase (HRP) catalyzed process. Chemosphere 58, pp: 1097–1105.23. Katheresan V. Kansedo J. Lau S. Y., 2018, Efficiency of various recent wastewater dye removal methods: A review. Journal of Environmental Chemical Engineering 6, pp: 4676–4697.24. Jin X., Li S., Long N., Zhang R., 2018, A robust and stable nano-biocatalyst by co-immobilization of chloroperoxidase and horseradish peroxidase for the decolorization of azo dyes. J Chem Technol Biotechnol , 93, pp: 489–497. 25. Yincan Z., Yan L., Xueyong G., Qiao W., Xiaoping X, 2017, Decolorization of Color Index Acid Orange 20 buffer solution using horseradish peroxidase immobilized on modified PAN-beads. RSC Adv., 7, pp: 18976. 26. Reactive Blue 160, http://www.worlddyevar_ety.com/react_ve-dyes/react_ve-blue-160.html 1/ Access time: 01 December, 2018.27. Roat C., Kadam A., Patel T., Dave S., 2016, Biodegradation of Diazo Dye, Reactive Blue 160 by Isolate Microbacterium sp. B12 Mutant: Identification of Intermediates by LC-MS. Int. J. Curr. Microbiol. App. Sci, 5(3), pp: 534-547.28. Patil A. V., Jadhav J. P., 2013, Evaluation of phytoremediation potential of Tagetes patula L. for the degradation of textile dye Reactive Blue 160 and assessment of the toxicity of degraded metabolites by cytogenotoxicity. Chemosphere 92, pp: 225–232. 29. Balapure K. H., Jain K., Chattaraj S., Bhatt N. S., Madamwar D., 2014, Co-metabolic degradation of diazo dye—Reactive blue 160 by enriched mixed cultures BDN. Journal of Hazardous Materials 279, 85–95.