Two potentially novel amylolytic enzyme specificities in the prokaryotic glycoside hydrolase α-amylase family GH57

Abstract

Glycoside hydrolase (GH) family 57 consists of more than 900 proteins from Archaea (roughly one-quarter) and Bacteria (roughly three-quarters), mostly from thermophiles. Fewer than 20 GH57 members have already been biochemically characterized as real, (almost exclusively) amylolytic enzymes. In addition to a recently described dual-specificity amylopullulanase-cyclomaltodextrinase, five enzyme specificities have been well established in the family--α-amylase, amylopullulanase, branching enzyme, 4-α-glucanotransferase and α-galactosidase--plus a group of the so-called α-amylase-like homologues probably without the enzyme activity. A (β/α)7-barrel succeeded by a bundle of a few α-helices forming the catalytic domain, and five conserved sequence regions (CSRs), are the main characteristics of family GH57. The main goal of the present bioinformatics study was to describe two novel groups within family GH57 that represent potential non-specified amylases (127 sequences mostly from Bacteria) and maltogenic amylases (12 sequences from Archaea). These were collected from sequence databases based on an indication of their biochemical characterization. Although both the non-specified amylases and the maltogenic amylases share the in silico identified catalytic machinery and predicted fold with the experimentally determined GH57 members, the two novel groups may define new GH57 subfamilies. They are distinguishable from the other, previously recognized, subfamilies by specific sequence features present especially in their CSRs (the so-called sequence fingerprints), also reflecting their own evolutionary histories.