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KMID : 0545120170270010179
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Journal of Microbiology and Biotechnology
2017 Volume.27 No. 1 p.179 ~ p.188
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Trametes villosa Lignin Peroxidase (TvLiP): Genetic and Molecular Characterization
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De Oliveira Carneiro Rita Terezinha
Lopes Maiza Alves
Silva Marilia Lordelo Cardoso
Da Silva Santos Veronica
De Souza Volnei Brito
De Sousa Aurizangela Oliveira
Pirovani Carlos Priminho
Koblitz Maria Gabriela Bello
Benevides Raquel Guimaraes
Goes-Neto Aristoteles
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Abstract
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White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the 7th day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.
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KEYWORD
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Lignin, ligninolytic enzymes, white-rot basidiomycetes
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