Introducing transglycosylation activity in a liquefying α‐amylase

FEBS Letters

López-Munguı́a, Agustı́n¹  Santamarı́a, Rosa I¹  Saab-Rincón, Gloria¹  Soberón, Xavier¹  del-Rı́o, Gabriel¹ 

[1] Instituto de Biotecnologı́a, UNAM, Apartado Postal 510-3, Cuernavaca, Morelos 62271, Mexico

关键词: α-Amylase;    Transglycosylation;    Hydrolysis;    Site-directed mutagenesis;    Reaction mechanism;    TAA;    taka amylase or alpha-amylase from Aspergillus oryzae;    ABST;    α-amylase from Bacillus stearothermophilus;    CGTase;    cyclodextrin glycosyltransferase from Bacillus circulans;    DNS;    3;    5-dinitrosalicylic acid;    MES;    2-(N-morpholino) ethanesulfonic acid;    MOPS;    3′-(N-porpholino) propanesulfonic acid;    G1;    glucose;    G2;    maltose;    G3;    maltotriose;    G4;    maltotetraose;    G5;    maltopentaose;    G6;    maltohexaose;    G7;    maltoheptaose;    PCR;    polymerase chain reaction;    TLC;    thin-layer chromatography;    HPLC;    high-pressure liquid chromatography;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    PDB;    Protein Databank;   

DOI  :  10.1016/S0014-5793(99)00671-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

By mutating Ala-289 by Phe or Tyr in the Bacillus stearothermophilus α-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, 17321–17329]. 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.

【 授权许可】

Unknown