Dividing the α-amylase family GH57 of starch hydrolases and related enzymes into subfamilies using evolutionary, clustering and functional criteria.

In the CAZy classification, four glycoside hydrolase families - GH13, GH57, GH119 and GH126 - have been designated as α-amylase families. The present study applied to a dataset of over 5000 family GH57 sequences, delivers a functionally balanced subdivision of the family GH57 into ten subfamilies. Eight of these subfamilies bear a specific enzyme activity: GH57_1, 4-α-glucanotransferases; GH57_2, amylopullulanases; GH57_3, α-amylases; GH57_4, α-galactosidases; GH57_5, α-glucan-branching enzymes; GH57_6, non-specified amylases; GH57_7, amylopullulanases-cyclomaltodextrinases; and GH57_8, maltogenic amylases. Two subfamilies, GH57_9 and GH57_10, not including any characterized members so far despite their conserved catalytic machinery, will deserve the community attention. Each subfamily is highlighted by its sequence fingerprints, through the logo of the five GH57 conserved sequence regions. The structural features are also compared with regard to domains complementing the catalytic module composed by the (β/α)7-barrel and the succeeding α-helical bundle. Characterized members in each subfamily display a strong agreement in their functional profile, indicating that the here proposed GH57 subfamily annotation results in functionally meaningful subsets. Moreover, several small groups of sequences still lacking sufficient sequence diversity and biochemical characterization did not integrate any of the created GH57 subfamilies so far; nevertheless, they could complete the overall GH57 subfamily picture in future.