The structures of the GH13_36 amylases from Eubacterium rectale and Ruminococcus bromii reveal subsite architectures that favor maltose production

• Published in: Amylase Vol. 4; no. 1; pp. 24 - 44
• Main Authors: Cockburn, Darrell W., Cerqueira, Filipe M., Bahr, Constance, Koropatkin, Nicole M
• Format: Journal Article
• Language: English
• Published: De Gruyter 31.12.2020
• Subjects: amylase; glycoside hydrolase family 13; microbiota; starch
• ISSN: 2450-9728; 2450-9728
• DOI: 10.1515/amylase-2020-0003

Bacteria in the human gut including and encode starch-active enzymes that dictate how these bacteria interact with starch to initiate a metabolic cascade that leads to increased butyrate. Here, we determined the structures of two predicted secreted glycoside hydrolase 13 subfamily 36 (GH13_36) enzymes: ErAmy13B complexed with maltotetraose from and RbAmy5 from The structures show a limited binding pocket extending from –2 through +2 subsites with limited possibilities for substrate interaction beyond this, which contributes to the propensity for members of this family to produce maltose as their main product. The enzyme structures reveal subtle differences in the +1/+2 subsites that may restrict the recognition of larger starch polymers by ErAmy13B. Our bioinformatic analysis of the biochemically characterized members of the GH13_36 subfamily, which includes the cell-surface GH13 SusG from , suggests that these maltogenic amylases (EC 3.2.1.133) are usually localized to the outside of the cell, display a range of substrate preferences, and most likely contribute to maltose liberation at the cell surface during growth on starch. A broader comparison between GH13_36 and other maltogenic amylase subfamilies explain how the activity profiles of these enzymes are influenced by their structures.