Introducing transglycosylation activity in a liquefying alpha-amylase.
By mutating Ala-289 by Phe or Tyr in the Bacillus stearothermophilus alpha-amylase, we induced this enzyme to perform alcoholytic reactions, a function not present in the wild-type enzyme. This residue was selected from homology analysis with neopullulanase, where the residue has been implicated in the control of transglycosylation [Kuriki et al. (1996) J. Biol. Chem. 271, 17321-173291. We made some inferences about the importance of electrostatic and geometrical modifications in the active site environment of the amylase to explain the behavior of the modified enzyme.
Saab-Rincón
G
G
Instituto de Biotecnología, UNAM, Cuernavaca, Morelos, Mexico.
del-Río
G
G
Santamaría
R I
RI
López-Munguía
A
A
Soberón
X
X
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D020134:Catalytic Domain; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016415:Sequence Alignment; D017421:Sequence Analysis; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
D016695:Glycosyltransferases; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; C058484:neopullulanase