Action of neopullulanase. Neopullulanase catalyzes both hydrolysis and transglycosylation at alpha-(1----4)- and alpha-(1----6)-glucosidic linkages.
The transglycosylation reaction catalyzed by neopullulanase was analyzed. Radioactive oligosaccharides were produced when the enzyme acted on maltotriose in the presence of [U-14C]glucose. Some of the radioactive oligosaccharides had only alpha-(1----4)-glucosidic linkages, but others were suggested to have alpha-(1----6)-glucosidic linkages. The existence of alpha-(1----6)-glucosidic linkages in the products from maltotriose with neopullulanase was proven by proton NMR spectroscopy and methylation analysis. We previously reported that the one active center of neopullulanase catalyzes the hydrolysis of alpha-(1----4)- and alpha-(1----6)-glucosidic linkages (Kuriki, T., Takata, H., Okada, S., and Imanaka, T. (1991) J. Bacteriol. 173,6147-6152). These facts proved that neopullulanase catalyzed all four types of reactions: hydrolysis of alpha-(1----4)-glucosidic linkage, hydrolysis of alpha-(1----6)-glucosidic linkage, transglycosylation to form alpha-(1----4)-glucosidic linkage, and transglycosylation to form alpha-(1----6)-glucosidic linkage. The four reactions are typically catalyzed by alpha-amylase, pullulanase, cyclomaltodextrin glucanotransferase, and 1,4-alpha-D-glucan branching enzyme, respectively. These four enzymes have some structural similarities to one other, but reactions catalyzed by the enzymes are considered to be distinctive: the four reactions are individually catalyzed by each of the enzymes. The experimental results obtained from the analysis of the reaction of the neopullulanase exhibited that the four reactions can be catalyzed in the same mechanism.
Takata
H
H
Biochemical Research Laboratories, Ezaki Glico Co., Ltd., Osaka, Japan.
Kuriki
T
T
Okada
S
S
Takesada
Y
Y
Iizuka
M
M
Minamiura
N
N
Imanaka
T
T
D015061:1,4-alpha-Glucan Branching Enzyme / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D002854:Chromatography, Paper; D005960:Glucosides / Q000378:metabolism*; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D006868:Hydrolysis; D007517:Isoamylase / Q000235:genetics / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D008969:Molecular Sequence Data; D009834:Oligo-1,6-Glucosidase / Q000235:genetics / Q000378:metabolism; D014312:Trisaccharides / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism; D000520:alpha-Glucosidases / Q000235:genetics / Q000378:metabolism
D005960:Glucosides; D014312:Trisaccharides; C008317:maltotriose; D005964:Glucosyltransferases; D015061:1,4-alpha-Glucan Branching Enzyme; C019963:cyclomaltodextrin glucanotransferase; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; D009834:Oligo-1,6-Glucosidase; C058484:neopullulanase; D000520:alpha-Glucosidases; C100181:pullulanase; D007517:Isoamylase