Cloning and Expression Analysis of Dye-decolorization Peroxidase Gene in Ganoderma lucidum
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Abstract:
To elucidate the potential role of dye-decolorizing peroxidase (DyP) gene in Ganoderma lucidum, G. lucidum ZJ-1 was evaluated using bioinformatics and expression analysis. To this end, the dye-decolorizing peroxidase genes GlDyP1 and GlDyP2 were cloned. The coding frame lengths were 1 488 bp and 1 461 bp, encoding 495 and 486 amino acids, respectively. The results of qRT-PCR analysis indicated that the expression levels of the two genes in the mycelium stage were significantly higher than in other stages, suggesting that GlDyP1 and GlDyP2 may be involved in the degradation of lignocellulose in the mycelium stage. The activity of GlDyP in G. lucidum was determined under different substrate cultures and dye induction conditions, and then the expression patterns of GlDyP1 and GlDyP2 were analyzed. The results attributed the highest enzyme activity of 7 330 U/L and 1,466 U/L to wood chips cultured in different substrates and methyl orange dye when induced by different dyes, respectively. GlDyP1 expression was highest in peanut shells, 6.5 times higher than that in sawdust. By contrast, GlDyP2 expression was highest in sawdust, 2 times higher than that in the other substrates. Compared with the control, the GlDyP1 and GlDyP2 genes showed the highest expression levels in reactive black 5 and trypan blue, with a 4.8-fold and 3.7-fold increase, respectively. These results indicate that GlDyP may participate in the degradation of lignocellulose and dyes by G. lucidum. These findings provide a foundation for exploring the physiological function of GlDyP in degrading lignin and dyes, and development of applications for GlDyP.
