ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
Purification and characterization of a laccase from the basidiomycete Funalia trogii (Berk.) isolated in Tanzania
Ferdinandi Patrick1, Godliving Mtui1*, Anthony Manoni Mshandete1, Gunnar Johansson2 and Amelia Kivaisi1
1Department of Molecular Biology and Biotechnology, University of Dar es Salaam, P. O. Box 35179 Dar es Salaam, Tanzania.
2Department of Biochemistry and Organic Chemistry, Uppsala University, P. O. Box 576, BMC SE75123, Uppsala, Sweden.
Accepted 25 February, 2017
Abstract
A lignolytic basidiomycete fungus, Funalia trogii (Berk.), was isolated from decayed wood in coastal Tanzania and cultivated in submerged culture. Initially screened crude enzyme filtrate showed complete rhemazol brilliant blue - R (RBBR) decolorization 2,2’-azino-bis (3-ethylbenzthiazoline)-6-sulfonate and guaiacol oxidation after 7 days of incubation. The fungal filtrate had maximum laccase activity of 593 U/ml after 15 days of incubation. A laccase was purified by anion exchange and size exclusion chromatography to good purity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF). The isolated main component had a molecular weight of ca 58 kDa as determined by MS and an isoelectric point (pI) of 3.8. The optimal pH and temperature range for the purified laccase were 4.0 - 5.0 and 50 - 70 ºC, respectively, using 2, 6-dimethoxyphenol as a substrate.
Key words: Funalia trogii, laccase, anion exchange chromatography, size exclusion chromatography, electrophoresis, isoelectric focusing.