Purification of a thermostable laccase from Leucaena leucocephala using a copper alginate entrapment approach and the application of the laccase in dye decolorization |
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| Institution: | 1. Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 570-752, South Korea;2. Bionano Systems Engineering Department, Chonbuk National University, Jeonju 561-756, South Korea;3. Department of Crop Science and Biotechnology, College of Agricultural and Life Science, Chonbuk National University, Jeonju 561-756, South Korea;4. Department of Molecular Biology, College of Natural Science, Chonbuk National University, Jeonju 561-756, South Korea;1. Strategic Water Infrastructure Lab, School of Civil, Mining and Environmental Engineering, University of Wollongong, Wollongong, NSW 2522, Australia;2. Smart Water Research Centre, Australian Rivers Institute, School of Environment, Griffith University, QLD 4222, Australia;3. Strategic Water Infrastructure Lab, School of Chemistry, University of Wollongong, Wollongong, NSW 2522, Australia;4. School of Civil, Environmental and Chemical Engineering, RMIT University, Melbourne, VIC 3001, Australia;5. Department of Civil Engineering, King Abdul Aziz University, Jeddah 21589, Saudi Arabia;1. Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, Belgrade, Serbia;2. Institute of Chemistry, Technology and Metallurgy-Center of Chemistry, University of Belgrade, Studentski trg 12-16, Belgrade, Serbia;1. Laboratory of Enzymology and Recombinant DNA Technology, Department of Microbiology, Maharshi Dayanand University, Rohtak 124001, Haryana, India;2. School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India;1. Industrial Biotechnology Laboratory, Department of Biochemistry, University of Agriculture, Faisalabad 38040, Pakistan;2. Department of Biochemistry, Government College Women University, Faisalabad, Pakistan;3. State Key Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;1. Institute of Sciences of Food Production, National Research Council of Italy (ISPA-CNR), Via G. Amendola 122/O, 70126, Bari, Italy;2. Institute of Biomembranes, Bioenergetics and Molecular Biotechnologies (IBIOM-CNR) Via Giovanni Amendola, 165/A, 70126, Bari, Italy |
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| Abstract: | Laccase from a tree legume, Leucaena leucocephala, was purified to homogeneity using a quick two-step procedure: alginate bead entrapment and celite adsorption chromatography. Laccase was purified 110.6-fold with an overall recovery of 51.0% and a specific activity of 58.5 units/mg. The purified laccase was found to be a heterodimer (∼220 kDa), containing two subunits of 100 and 120 kDa. The affinity of laccase was found to be highest for catechol and lowest for hydroquinone, however, highest Kcat and Kcat/Km were obtained for hydroquinone. Purified laccase exhibited pH and temperature optima of 7.0 and 80 °C, respectively. Mn2+, Cd2+, Fe2+, Cu2+ and Na+ activated laccase while Ca2+ treatment increased laccase activity up to 3 mM, beyond which it inhibited laccase. Co2+, Hg2+, DTT, SDS and EDTA showed an inhibition of laccase activity. The Leucaena laccase was found to be fairly tolerant to organic solvents; upon exposure for 1 h individually to 50% (v/v) each of ethanol, DMF, DMSO and benzene, more than 50% of the activity was retained, while in the presence of 50% (v/v) each of methanol, isopropanol and chloroform, a 40% residual activity was observed. The purified laccase efficiently decolorized synthetic dyes such as indigocarmine and congo red in the absence of any redox mediator. |
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| Keywords: | Celite chromatography Copper alginate Dye decolorization Laccase |
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