Trametes villosa lignin peroxidase (TvLiP): genetic and molecular characterization.

White-rot basidiomycetes are the organisms that decompose lignin most efficiently and Trametes villosa is a promising species for the ligninolytic enzymes production. There are several publications on Trametes villosa applications for lignin degradation regarding the expression and secretion of laccase (Lac) and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breaking down 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 a strain of Trametes villosa CCMB561 from the Brazilian semi-arid 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 RBBR. 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 medium that best 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.