Abstract
Two chimeric genes, XynA-Bs-Glu-1 and XynA-Bs-Glu-2, encoding Aspergillus sulphureus β-xylanase (XynA, 26 kDa) and Bacillus subtilis β-1,3-1,4-glucanase (Bs-Glu, 30 kDa), were constructed via in-fusion by different linkers and expressed successfully in Pichia pastoris. The fusion protein (50 kDa) exhibited both β-xylanase and β-1,3-1,4-glucanase activities. Compared with parental enzymes, the moiety activities were decreased in fermentation supernatants. Parental XynA and Bs-Glu were superior to corresponding moieties in each fusion enzymes because of lower Kn higher kcat. Despite some variations, common optima were generally 50°C and pH 3.4 for the XynA moiety and parent, and 40°C and pH 6.4 for the Bs-Glu counterparts. Thus, the fusion enzyme XynA-Bs-Glu-1 and XynA-Bs-Glu-2 were bifunctional.
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Abbreviations
- BMGY:
-
buffered glycerol-complex medium
- BMMY:
-
buffered methanol-complex medium
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Qiao, J., Cao, Y. In-fusion expression and characterization of β-xylanase and β-1,3-1,4-glucanase in Pichia pastoris . Biologia 67, 649–653 (2012). https://doi.org/10.2478/s11756-012-0056-3
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DOI: https://doi.org/10.2478/s11756-012-0056-3