Abstract
A salt-tolerant exo-β-1,3-glucosidase (BGL_MK86) was cloned from the xerophilic mold Aspergillus chevalieri MK86 and heterologously expressed in A. oryzae. Phylogenetic analysis suggests that BGL_MK86 belongs to glycoside hydrolase family 5 (aryl-phospho-β-D-glucosidase, BglC), and exhibits D-glucose tolerance. Recombinant BGL_MK86 (rBGL_MK86) exhibited 100-fold higher expression than native BGL_MK86. rBGL_MK86 was active over a wide range of NaCl concentrations [0%–18% (w/v)] and showed increased substrate affinity for p-nitrophenyl-β-D-glucopyranoside (pNPBG) and turnover number (kcat) in the presence of NaCl. The enzyme was stable over a broad pH range (5.5–9.5). The optimum reaction pH and temperature for hydrolysis of pNPBG were 5.5 and 45 °C, respectively. rBGL_MK86 acted on the β-1,3-linked glucose dimer laminaribiose, but not β-1,4-linked or β-1,6-linked glucose dimers (cellobiose or gentiobiose). It showed tenfold higher activity toward laminarin (a linear polymer of β-1,3 glucan) from Laminaria digitata than laminarin (β-1,3/β-1,6 glucan) from Eisenia bicyclis, likely due to its inability to act on β-1,6-linked glucose residues. The β-glucosidase retained hydrolytic activity toward crude laminarin preparations from marine biomass in moderately high salt concentrations. These properties indicate wide potential applications of this enzyme in saccharification of salt-bearing marine biomass.
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Acknowledgements
This work was supported by the Japan Society for the Promotion of Science KAKENHI [grant number, 20K05808]. We thank James Allen, DPhil, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.
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Funding was supported by Japan Society for the Promotion of Science KAKENHI (20K05808).
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HS and DS: Conceptualization, Molecular Biology, Data collection, Writing–original draft preparation. YK, ST, and MD: Conceptualization, Reviewing–original draft. ST: Conceptualization, Bioinformatics, Writing –Reviewing and Editing. All authors reviewed the final version of the manuscript.
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Senba, H., Saito, D., Kimura, Y. et al. Heterologous expression and characterization of salt-tolerant β-glucosidase from xerophilic Aspergillus chevalieri for hydrolysis of marine biomass. Arch Microbiol 205, 310 (2023). https://doi.org/10.1007/s00203-023-03648-z
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DOI: https://doi.org/10.1007/s00203-023-03648-z