X-ray structure of Novamyl, the five-domain "maltogenic" alpha-amylase from Bacillus stearothermophilus: maltose and acarbose complexes at 1.7A resolution

Biochemistry. 1999 Jun 29;38(26):8385-92. doi: 10.1021/bi990256l.


The three-dimensional structure of the Bacillus stearothermophilus "maltogenic" alpha-amylase, Novamyl, has been determined by X-ray crystallography at a resolution of 1.7 A. Unlike conventional alpha-amylases from glycoside hydrolase family 13, Novamyl exhibits the five-domain structure more usually associated with cyclodextrin glycosyltransferase. Complexes of the enzyme with both maltose and the inhibitor acarbose have been characterized. In the maltose complex, two molecules of maltose are found in the -1 to -2 and +2 to +3 subsites of the active site, with two more on the C and E domains. The C-domain maltose occupies a position identical to one previously observed in the Bacillus circulans CGTase structure [Lawson, C. L., et al. (1994) J. Mol. Biol. 236, 590-600], suggesting that the C-domain plays a genuine biological role in saccharide binding. In the acarbose-maltose complex, the tetrasaccharide inhibitor acarbose is found as an extended hexasaccharide species, bound in the -3 to +3 subsites. The transition state mimicking pseudosaccharide is bound in the -1 subsite of the enzyme in a 2H3 half-chair conformation, as expected. The active site of Novamyl lies in an open gully, fully consistent with its ability to perform internal cleavage via an endo as opposed to an exo activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acarbose
  • Amino Acid Sequence
  • Binding Sites
  • Carrier Proteins / chemistry
  • Carrier Proteins / metabolism
  • Computer Simulation
  • Crystallography, X-Ray
  • Enzyme Inhibitors / chemistry
  • Geobacillus stearothermophilus / enzymology*
  • Macromolecular Substances
  • Maltose / chemistry*
  • Maltose / metabolism
  • Maltose-Binding Proteins
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Trisaccharides / chemistry*
  • alpha-Amylases / antagonists & inhibitors
  • alpha-Amylases / chemistry*
  • alpha-Amylases / metabolism


  • Carrier Proteins
  • Enzyme Inhibitors
  • Macromolecular Substances
  • Maltose-Binding Proteins
  • Trisaccharides
  • Maltose
  • alpha-Amylases
  • Acarbose

Associated data

  • PDB/1QHO
  • PDB/1QHP