Insight into the substrate specificity change caused by the Y227H mutation of α-glucosidase III from the European honeybee (Apis mellifera) through molecular dynamics simulations

• Published in: PloS one Vol. 13; no. 6; p. e0198484
• Main Authors: Na Ayutthaya, Pratchaya Pramoj, Chanchao, Chanpen, Chunsrivirot, Surasak
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 04.06.2018; Public Library of Science (PLoS)
• Subjects: alpha-Glucosidases - genetics; alpha-Glucosidases - metabolism; Animals; Apis mellifera; Bees; Binding; Binding Sites; Biochemistry; Biology; Biology and Life Sciences; Catalytic Domain; Computer simulation; Enzymes; Food industry; Free energy; Glucosidase; Homology; Hydrogen Bonding; Hydrolysis; Insect Proteins - genetics; Insect Proteins - metabolism; Isoforms; Malt; Maltose; Maltose - chemistry; Maltose - metabolism; Mechanics; Molecular chains; Molecular dynamics; Molecular Dynamics Simulation; Mutation; Peptides; Physical Sciences; Polymorphism, Single Nucleotide; Proteins; Substrate Specificity; Substrates; Sucrose; Sucrose - chemistry; Sucrose - metabolism; Sugar; Thermodynamics; α-Glucosidase; Thailand; Bangkok Thailand
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0198484

Honey from the European honeybee, Apis mellifera, is produced by α-glucosidases (HBGases) and is widely used in food, pharmaceutical, and cosmetic industries. Categorized by their substrate specificities, HBGases have three isoforms: HBGase I, II and III. Previous experimental investigations showed that wild-type HBGase III from Apis mellifera (WT) preferred sucrose to maltose as a substrate, while the Y227H mutant (MT) preferred maltose to sucrose. This mutant can potentially be used for malt hydrolysis because it can efficiently hydrolyze maltose. In this work, to elucidate important factors contributing to substrate specificity of this enzyme and gain insight into how the Y227H mutation causes substrate specificity change, WT and MT homology models were constructed, and sucrose/maltose was docked into active sites of the WT and MT. AMBER14 was employed to perform three independent molecular dynamics runs for these four complexes. Based on the relative binding free energies calculated by the MM-GBSA method, sucrose is better than maltose for WT binding, while maltose is better than sucrose for MT binding. These rankings support the experimentally observed substrate specificity that WT preferred sucrose to maltose as a substrate, while MT preferred maltose to sucrose, suggesting the importance of binding affinity for substrate specificity. We also found that the Y227H mutation caused changes in the proximities between the atoms necessary for sucrose/maltose hydrolysis that may affect enzyme efficiency in the hydrolysis of sucrose/maltose. Moreover, the per-residue binding free energy decomposition results show that Y227/H227 may be a key residue for preference binding of sucrose/maltose in the WT/MT active site. Our study provides important and novel insight into the binding of sucrose/maltose in the active site of Apis mellifera HBGase III and into how the Y227H mutation leads to the substrate specificity change at the molecular level. This knowledge could be beneficial in the design of this enzyme for increased production of desired products.