Pattern of action of Bacillus stearothermophilus neopullulanase on pullulan.
The action of neopullulanase from Bacillus stearothermophilus on many oligosaccharides was tested. The enzyme hydrolyzed not only alpha-(1----4)-glucosidic linkages but also specific alpha-(1----6)-glucosidic linkages of several branched oligosaccharides. When pullulan was used as a substrate, panose, maltose, and glucose, in that order, were produced as final products at a final molar ratio of 3:1:1. According to these results, we proposed a model for the pattern of action of neopullulanase on pullulan as follows. In the first step, the enzyme hydrolyzes only alpha-(1----4)-glucosidic linkages on the nonreducing side of alpha-(1----6) linkages of pullulan and produces panose and several intermediate products composed of some panose units. In the second step, taking 6(2)-O-alpha-(6(3)-O-alpha-glucosyl-maltotriosyl)-maltose as an example of one of the intermediate products, the enzyme hydrolyzes either alpha-(1----4) (the same position as that described above) or alpha-(1----6) linkages and produces panose or 6(3)-O-alpha-glucosyl-maltotriose plus maltose, respectively. In the third step, the alpha-(1----4) linkage of 6(3)-O-alpha-glucosyl-maltotriose is hydrolyzed by the enzyme, and glucose and another panose are produced. To confirm the model of the pattern of action, we extracted intermediate products produced from pullulan by neopullulanase and analyzed the structures by glucoamylase, pullulanase, and neopullulanase analyses. The experimental results supported the above-mentioned model of the pattern of action of neopullulanase on pullulan.
Imanaka
T
T
Department of Fermentation Technology, Faculty of Engineering, Osaka University, Japan.
Kuriki
T
T
D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005936:Glucans / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007700:Kinetics; D009844:Oligosaccharides / Q000378:metabolism; D013379:Substrate Specificity
D005936:Glucans; D009844:Oligosaccharides; C009109:pullulan; D006026:Glycoside Hydrolases; C058484:neopullulanase