Biotechnology & Biotechnological Equipment (Jan 2020)
Molecular cloning and bioinformatics analyses of a GH3 beta-glucosidase from Auricularia heimuer
Abstract
The present study reports the isolation and analysis of a beta-glucosidase gene (accession number MN816859) from the fungus Auricularia heimuer. The Aa-bgl gene comprises 2583 bp and encodes a putative polypeptide of 860 amino acids. BlastP analysis revealed conserved glycoside hydrolase family 3 domains. Recombinant plasmid pET-32a-bgl was constructed to overexpress the target protein in Escherichia coli BL21, and sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis revealed target bands between 100 kDa and 135 kDa, consistent with the expected molecular weight. Additionally, the transcription levels of Aa-bgl at different developmental stages were investigated, and the results showed that the expression levels increased with the maturation of the fruiting bodies. In mature fruiting bodies, the levels were 6.69-fold higher than in controls (mycelium at 6 days). We therefore speculate that Aa-bgl may be correlated with development in A. heimuer. The findings provide a theoretical basis for studying the function of this beta-glucosidase in future work.
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