Herein we demonstrate how a refined sequenced-based approach can be used to improve the T/H ratio (i.e. enhance transglycosylation capability) in a variety of predominantly hydrolytic GHs. This strategy involves replacing an enzyme's most conserved residues by structural analogues (e.g., Tyr into Phe, Asp into Asn). The approach does not require structural or mechanistic knowledge of the GH. However, for enzymes where catalytic residues and residues distant (>10 Å) from the -1 subsite 23 are known or can be predicted, those are not mutated. Moreover, any conserved glycines and prolines are excluded . Using this strategy, we demonstrate successful application to GHs from families 2, 10, 20, 29 and 51. This approach is i) fast, requiring the generation and analysis of ≤ 10 variants per enzyme; ii) generic, being applicable to a variety of glycosidic bond-forming reactions; iii) procures highly efficient transglycosylases; and iv) allows for mutation transfer.