Available online at http://ajol.info/index.php/ijbcs Int. J. Biol. Chem. Sci. 6(2): 782-791, April 2012 ISSN 1991-8631

Original Paper

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Purification and partial characterization of laccase from Lachnocladium sp.
P. A. WUYEP1, O. N. UME 2, M. T. BAKARE-ODUNOLA 2*, A. J. NOK 3, H. M. INUWA 3 and N. B. AFOLABI-BALOGUN 4
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Department of Biological Sciences, Ahmadu Bello University, Zaria, Nigeria. Department of Pharmaceutical and Medicinal Chemistry, Ahmadu Bello University, Zaria, Nigeria. 3 Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria. 4 Center for Biotechnology Research and Traininig. Ahmadu Bello University, Zaria, Nigeria. * Corresponding author, E-mail: mojitaibat@yahoo.com; Tel.:+2348035896043

1

ABSTRACT
Laccase, a multicopper oxidase that catalyzes the oxidation of various aromatics, particularly phenolic substrates, e.g. hydroquinones guaiacol, 2,6-dimethoxyphenol or phenylene diamine, was purified and partially characterised from culture filtrates of a white rot fungus, Lachnocladium sp. This enzyme was purified by anion exchange and gel filtration chromatography. Laccase activity was determined using ABTS (2, 2’-azinobis-(3-ethylbenzthiazoline)-6-sulphonic acid) substrate. The culture filtrate had maximum laccase activity of 1.62 U/ml after 14 days of incubation. The purified laccase had an optimum temperature of 50 oC and its optimum pH was 6 for ABTS. The activity of this enzyme was enhanced by Fe2+, Cu2+, Zn2+and Ca2+, and was inhibited by EDTA and sodium iodide. Laccase from Lachnocladium sp. had a Km of 0.119 mM and a Vmax of 0.313 U. © 2012 International Formulae Group. All rights reserved. Keywords: Lachnocladium sp., anion exchange chromatography, gel filtration chromatography, ABTS, DMP.

INTRODUCTION Laccases (EC 1.10.3.2, p-diphenol: dioxygen oxidoreductase) belong to the