Function and structure of GH13_31 α-glucosidase with high α-(1→4)-glucosidic linkage specificity and transglucosylation activity

FEBS Lett. 2018 Jul;592(13):2268-2281. doi: 10.1002/1873-3468.13126. Epub 2018 Jun 20.

Abstract

α-Glucosidase hydrolyzes α-glucosides and transfers α-glucosyl residues to an acceptor through transglucosylation. In this study, GH13_31 α-glucosidase BspAG13_31A with high transglucosylation activity is reported in Bacillus sp. AHU2216 and biochemically and structurally characterized. This enzyme is specific to α-(1→4)-glucosidic linkage as substrates and transglucosylation products. Maltose is the most preferred substrate. Crystal structures of BspAG13_31A wild-type for the substrate-free form and inactive acid/base mutant E256Q in complexes with maltooligosaccharides were solved at 1.6-2.5 Å resolution. BspAG13_31A has a catalytic domain folded by an (β/α)8 -barrel. In subsite +1, Ala200 and His203 on β→α loop 4 and Asn258 on β→α loop 5 are involved in the recognition of maltooligosaccharides. Structural basis for specificity of GH13_31 enzymes to α-(1→4)-glucosidic linkage is first described.

Keywords: glycoside hydrolase family 13; transglycosylation; α-glucosidase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus / enzymology*
  • Bacillus / genetics
  • Carbohydrate Metabolism / genetics
  • Carbohydrate Sequence / physiology
  • Catalytic Domain / genetics
  • Glucosides / chemistry
  • Glucosides / metabolism*
  • Glycosylation
  • Hydrolysis
  • Kinetics
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Structure-Activity Relationship
  • Substrate Specificity / genetics
  • alpha-Glucosidases / chemistry*
  • alpha-Glucosidases / genetics
  • alpha-Glucosidases / metabolism*

Substances

  • Glucosides
  • alpha-Glucosidases

Associated data

  • GENBANK/AEN90673.1
  • GENBANK/BAE48285.1
  • GENBANK/BAA12704.1
  • GENBANK/BAL49684.1
  • GENBANK/BAE79634.1
  • GENBANK/AAV42157.1
  • GENBANK/CAA37583.1
  • GENBANK/CAB15461.1
  • GENBANK/CAA54266.1