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• W2896677240 abstract "In this study, computation-aided design on the basis of structural analysis was employed to rationally identify a highly dynamic C-terminal region that regulates the stability, expression level, and activity of a GH2 fungal glucuronidase from Aspergillus oryzae Li-3 (PGUS). Then, four mutants with a precisely truncated C-terminal region in different lengths were constructed; among them, mutant D591–604 with a 3.8-fold increase in half-life at 65 °C and a 6.8 kJ/mol increase in Gibbs free energy showed obviously improved kinetic and thermodynamic stability in comparison to PGUS. Mutants D590–604 and D591–604 both showed approximately 2.4-fold increases in the catalytic efficiency kcat/Km and 1.8-fold increases in the expression level. Additionally, the expression level of PGUS was doubled through a C-terminal region swap with bacterial GUS from E. coli (EGUS). Finally, the robust PGUS mutants D590–604 and D591–604 were applied in the preparation of glycyrrhetinic acid with 4.0- and 4.4-fold increases in concentration through glycyrrhizin hydrolysis by a fed-batch process." @default.
• W2896677240 created "2018-10-26" @default.
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• W2896677240 date "2018-10-08" @default.
• W2896677240 modified "2023-10-14" @default.
• W2896677240 title "Computation-Aided Rational Deletion of C-Terminal Region Improved the Stability, Activity, and Expression Level of GH2 β-Glucuronidase" @default.
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• W2896677240 doi "https://doi.org/10.1021/acs.jafc.8b03449" @default.
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• W2896677240 hasPublicationYear "2018" @default.
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