Design of mutants for enhanced thermostability of β-glycosidase BglY from Thermus thermophilus

► Three rational and semi-rational design approaches were employed to engineer BglY. ► Five beneficial mutations lead to increased thermostability of BglY. ► The t1/2 of mutant HF5 increases to 4.7-fold of that of the wild-type. ► Substrate inhibition is completely released mainly due to the A285Y mutation.