Trametes villosa Lignin Peroxidase (TvLiP): Genetic and Molecular Characterization

Abstract

White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and is a promising species for ligninolytic enzyme production. There are several publications on 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 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 7 day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the gene median region covering approximately 50% of the gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the mRNA; and the close phylogenetic relationship between TvLiP and LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

Draft genome sequence of Trametes villosa (Sw.) Kreisel CCMB561, a tropical white-rot Basidiomycota from the semiarid region of Brazil

“…Trametes villosa (Sw.) Kreisel is a common species in the Brazilian semiarid region [3] . It is a good producer of the three important ligninolytic enzymes: Laccase (Lac) [4] , Manganese Peroxidase (MnP) [5] and Lignin Peroxidase (LiP) [6] , demonstrating its high potential for biotechnological applications. However, little is known about the function and structure of T. villosa genes, which requires detailed investigation.…”

“…Nowadays, ligninolytic enzymes of white-rot fungi have been broadly studied for their potential applications in a wide range of industrial bioprocesses such as decolorization of industrial dyes, the pulp bleaching of paper, textile industry and the degradation of organopollutants [9] . Furthermore, T. villosa simultaneously produces LiP, MnP and Lac [5] , [6] , [10] whereas other lignin decay fungi produce only one or two of these ligninolytic enzymes simultaneously [11] , [12] . Thus, a species able to produce the three ligninolytic enzymes in the same bath culture is highly desirable for biotechnological applications [6] .…”

“…Furthermore, T. villosa simultaneously produces LiP, MnP and Lac [5] , [6] , [10] whereas other lignin decay fungi produce only one or two of these ligninolytic enzymes simultaneously [11] , [12] . Thus, a species able to produce the three ligninolytic enzymes in the same bath culture is highly desirable for biotechnological applications [6] .…”

“…Although T. villosa CCMB561 commonly exists as an efficient wood decomposer, with a lignocellulolytic enzyme system mainly comprising laccases, lignin peroxidases and Mn-dependent peroxidases as well as a series of CAZymes [5] , [6] , [16] , there are limited data about their synthesis, genetic coding and regulation. In the context of basic research, the genome sequencing of T. villosa CCMB561 presented herein will enrich the portfolio of potential genes, enzymes and pathways involved in the lignin degradation processes of the white-rot fungi.…”

Hybrid Assembly Improves Genome Quality and Completeness of Trametes villosa CCMB561 and Reveals a Huge Potential for Lignocellulose Breakdown

“…Trametes villosa is an important species of the genus Trametes, and its potential to produce laccases, peroxidases, and cellulases in different lignocellulosic substrates has already been demonstrated at the small-scale biochemical level [29][30][31][32]. Regarding the strain T. villosa CCMB561, biochemical assays have shown its potential to produce ligninolytic enzymes using sugarcane bagasse as a substrate in different temperatures (from 20 • C to 33 • C) and pHs (from 4.60 to 9.38) [29,32].…”