Action pattern and subsite mapping of Bacillus licheniformis alpha-amylase (BLA) with modified maltooligosaccharide substrates.
This study represents the first characterisation of the substrate-binding site of Bacillus licheniformis alpha-amylase (BLA). It describes the first subsite map, namely, number of subsites, apparent subsite energies and the dual product specificity of BLA. The product pattern and cleavage frequencies were determined by high-performance liquid chromatography, utilising a homologous series of chromophore-substituted maltooligosaccharides of degree of polymerisation 4-10 as model substrates. The binding region of BLA is composed of five glycone, three aglycone-binding sites and a 'barrier' subsite. Comparison of the binding energies of subsites, which were calculated with a computer program, shows that BLA has similarity to the closely related Bacillus amyloliquefaciens alpha-amylase.
Kandra
Lili
L
University of Debrecen, Faculty of Sciences, Department of Biochemistry, P.O. Box 55, H-4010, Debrecen, Hungary. kandra@tigris.klte.hu
Gyémánt
Gyöngyi
G
Remenyik
Judit
J
Hovánszki
György
G
Lipták
András
A
D001407:Bacillus / Q000201:enzymology*; D001597:Benzylidene Compounds / Q000737:chemistry; D001665:Binding Sites; D002240:Carbohydrate Sequence; D008320:Maltose / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000737:chemistry; D009716:Numerical Analysis, Computer-Assisted; D013379:Substrate Specificity; D000516:alpha-Amylases / Q000737:chemistry* / Q000378:metabolism*
D001597:Benzylidene Compounds; D009596:Nitrophenols; D008320:Maltose; D000516:alpha-Amylases