Identification of wild Wolfiporia cocos and study on its lignocellulolytic enzymes

Abstract

In order to reveal the effects of Wolfiporia cocos lignocellulolytic enzymes and culture methods on its main enzymes, the main lignocellulolytic enzymes of W. cocos were determined in this study. The microscopic observation of the culture characteristics of the wild W. cocos strains was carried out, three pairs of primers were used for PCR amplification to carry out phylogenetic identification, and the dominant strain YX1 was screened by qualitative culture and finally the activities of cellulase, hemicellulase and ligninolytic enzymes under different conditions were determined by microplate reader. The results were as follows:（1）W. cocos had mycelium, fruiting body and sclerotium three morphological characteristics.（2）PCR obtained rDNA-ITS sequence of 1 652 bp, ribosomal large subunit sequence of 660 bp and translation elongation factor sequence of 545 bp, and submitted to NCBI，accession numbers were ON129554, ON129553, and ON155840, respectively.（3）The highest secretion of exo-β-glucanase(CBH), endo-β-glucanase(EG) and β-glucosidase(BGL) in the presence or absence of sawdust was 16-17 U·mL-1, 32-35 U·mL-1, 36-37 U·mL-1；The maximum secretion of xylanase, mannanase and α-glucosidase was 28-38 U·mL-1, 280-342 U·mL-1, 9-11 U·mL-1. The three ligninolytic enzymes manganese peroxidase (MnP), laccase, and lignin peroxidase (LiP) had weak enzymatic activities in four different cultures, the magnitude of enzymatic activities in lignocellulases were in the order of mannanase > xylanase > BGL > EG > CBH >α-glucosidase > LiP > MnP > laccase, and there were significant differences between cellulase and hemicellulase enzymatic activities (P<0.05). In conclusion, this study combines morphological and molecular identification to clarify the taxonomic status of YX1, which has both relationship and genetic gap with brown rot fungi, and provides a basic enzymatic reference for the degradation mechanism of the lignocellulolytic enzymes system produced by W. cocos.

Keywords

• wolfiporia cocos
• its
• cellulase
• hemicellulase
• ligninolytic enzymes