Engineering the cytokinin-glucoside specificity of the maize β-d-glucosidase Zm-p60.1 using site-directed random mutagenesis

► Zm-p60.1 β-d-glucosidase has been engineered to discriminate between cZOG and tZOG. ► The highest preference for cZOG was shown by P372T/W373K/M376L. ► cZOG is a more favorable substrate for wild-type Zm-p60.1. ► Site-directed random mutagenesis could be used to modulate cytokinin metabolism.