Effects of lignocellulolytic enzymes activities under different culture conditions from Wolfiporia cocos
Abstract
Objective: In order to explore the degradation mechanism of Wolfiporia cocos fungus lignocellulose, reveal the main enzymes of poria cocos lignocellulose system and their relationship with culture methods, and explore the production and application of lignocellulose-related enzyme resources. Method: Poria cocos strain YX2 collected in the field was observed microscopically to understand its culture characteristics. Its DNA was extracted by fungal kit and then amplified by PCR. After the obtained products were compared by BLAST, the phylogenetic tree of brown rot fungus was constructed by using biological analysis software ClustalX and building Phylogenetic Trees from Neighbor-Joining with MEGA. The activities of cellulase, hemicellulase and lignin-degrading enzymes were determined by MicroplateReader, and the magnitude of the nine enzyme activities were calculated. The maximum secretion of exo- -glucanase, endo- -glucanase and -glucosidase in the cellulase group was 16-17 U/mL, 32-35 U/mL and 36-37 U/mL, respectively, and the maximum secretion of xylanase, mannanase and α-glucosidase in the hemicellulase group was 28-38 U/mL, 280-342 U/mL and 9-11 U/mL, respectively, under the conditions of treatment with or without the addition of pine wood chips. 280-342U/mL, and 9-11U/mL, respectively.The maximum secretion of MnP, Laccase and LiP, which are lignin degrading enzymes, in four different culture solutions: A. without were 0.015U/mL, 0.031U/mL and 0.017U/mL, respectively; B. with Mn2+ were 0.081U/mL, 0.032U/mL and 0.109U/mL, respectively; C. with The highest secretion of wood chips was 0.026U/mL, 0.025U/mL, 0.105U/mL, respectively; D. The highest secretion of 2,6DMP was 0.025U/mL, 0.029U/mL, 0.067U/mL, respectively. Conclusion: Through the combination of morphological and molecular biological identification of Poria cocos, the taxonomic status of Poria YX2 was clarified, and the brown rot fungus in There is both a connection and a genetic gap in the affinity. The size of the enzymatic activity in lignocellulase in the order of mannanase > xylanase > -glucosidase > endo- -glucanase > exo- -glucanase > -glucosidase > LiP > MnP > Laccase, and to provide a basic enzymatic reference for the study of the mechanism of action of the lignocellulase system produced by Porphyromonas.
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