Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 30172
Extraction in Two-Phase Systems and Some Properties of Laccase from Lentinus polychrous

Authors: K. Ratanapongleka, J. Phetsom

Abstract:

Extraction of laccase produced by L. polychrous in an aqueous two-phase system, composed of polyethylene glycol and phosphate salt at pH 7.0 and 250C was investigated. The effect of PEG molecular weight, PEG concentration and phosphate concentration was determined. Laccase preferentially partitioned to the top phase. Good extraction of laccase to the top phase was observed with PEG 4000. The optimum system was found in the system containing 12% w/w PEG 4000 and 16% w/w phosphate salt with KE of 88.3, purification factor of 3.0-fold and 99.1% yield. Some properties of the enzyme such as thermal stability, effect of heavy metal ions and kinetic constants were also presented in this work. The thermal stability decreased sharply with high temperature above 60 0C. The enzyme was inhibited by Cd2+, Pb2+, Zn2+ and Cu2+. The Vmax and Km values of the enzyme were 74.70 μmol/min/ml and 9.066 mM respectively.

Keywords: Aqueous Two Phase System, Laccase, Lentinuspolychrous,

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1077651

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1573

References:


[1] S.R. Couto, Decolouration of industrial azo dyes by crude laccase from Trametes hirsuta, Journal of Hazardous Materials 148(2007) 768-770.
[2] R. Khlifi, L. Belbahri, S. Woodward, M. Ellouz, A. Dhouib, S. Sayadi, T. Mechichi, Decolourization and detoxification of textile industry wastewater by the laccase-mediator system, Journal of Hazardous Materials 175(2010) 802-808.
[3] T. Saito, K. Kato, Y. Yokogawa, M. Nishida, N. Yamashita, Detoxification of bisphenol A and nonylphenol by purified extracellular laccase from a fungus isolated from soil, Journal of Bioscience and Bioengineering 98(2004) 64-66.
[4] M. Lund, C. Felby, Wet strength improvement of unbleached kraft pulp through laccase catalyzed oxidation, Enzyme and Microbial Technology 28(2001) 760-765.
[5] PazarlIog, N.K. lu, M. Sariisik, A. Telefoncu, Laccase: production by Trametes versicolor and application to denim washing, Process Biochemistry 40(2005) 1673-1678.
[6] E. Selinheimo, K. Kruus, J. Buchert, A. Hopia, K. Autio, Effects of laccase, xylanase and their combination on the rheological properties of wheat doughs, Journal of Cereal Science 43(2006) 152-159.
[7] R.C. Minussi, G.M. Pastore, N. DurÓ╣ün, Potential applications of laccase in the food industry, Trends in Food Science & Technology 13 205-216.
[8] R. Mustafa, L. Muniglia, B. Rovel, M. Girardin, Phenolic colorants obtained by enzymatic synthesis using a fungal laccase in a hydroorganic biphasic system, Food Research International 38(2005) 995- 1000.
[9] D. Litthauer, M.J. van Vuuren, A. van Tonder, F.W. Wolfaardt, Purification and kinetics of a thermostable laccase from Pycnoporus sanguineus (SCC 108), Enzyme and Microbial Technology 40(2007) 563-568.
[10] A. Veide, A.-L. Smeds, S.-O. Enfors, A Process for Large Scale Isolation of b-galactosidase from E. coli in an Aqueous Two- Phase System, Biotechnology and Bioengineering 25(1983) 1789-1800.
[11] H. Walter, G. Johansson, Aqueous Two-Phase Systems. Methods in Enzymology, Academic Press, New York, 1994, 228 pp.
[12] M. van Berlo, K.C.A.M. Luyben, L.A.M. van der Wielen, Poly(ethylene glycol)-salt aqueous two-phase systems with easily recyclable volatile salts, Journal of Chromatography B: Biomedical Sciences and Applications 711(1998) 61-68.
[13] M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry 72(1976) 248-254.
[14] H.S. Mohamadi, E. Omidinia, R. Dinarvand, Evaluation of recombinant phenylalanine dehydrogenase behavior in aqueous two-phase partitioning, Process Biochemistry 42(2007) 1296-1301.
[15] I. Yücekan, S. Önal, Partitioning of invertase from tomato in poly(ethylene glycol)/sodium sulfate aqueous two-phase systems, Process Biochemistry 46(2011) 226-232.
[16] S. Kalme, S. Jadhav, M. Jadhav, S. Govindwar, Textile dye degrading laccase from Pseudomonas desmolyticum NCIM 2112, Enzyme and Microbial Technology 44(2009) 65-71.
[17] S. Sadhasivam, S. Savitha, K. Swaminathan, F.-H. Lin, Production, purification and characterization of mid-redox potential laccase from a newly isolated Trichoderma harzianum WL1, Process Biochemistry 43(2008) 736-742.
[18] K. Murugesan, Y.-M. Kim, J.-R. Jeon, Y.-S. Chang, Effect of metal ions on reactive dye decolorization by laccase from Ganoderma lucidum, Journal of Hazardous Materials 168(2009) 523-529.
[19] M. Lorenzo, D. Moldes, S. RodrIguez Couto, M.A. SanromÓ╣ün, Inhibition of laccase activity from Trametes versicolor by heavy metals and organic compounds, Chemosphere 60(2005) 1124-1128.G. O. Young, "Synthetic structure of industrial plastics (Book style with paper title and editor)," in Plastics, 2nd ed. vol. 3, J. Peters, Ed. New York: McGraw-Hill, 1964, pp. 15-64.