Transglycosylation of glycosyl residues to cyclic tetrasaccharide by Bacillus stearothermophilus cyclomaltodextrin glucanotransferase using cyclomaltodextrin as the glycosyl donor.
Cyclomaltodextrin glucanotransferase (EC 2.4.1.19, abbreviated as CGTase) derived from Bacillus stearothermophilus produced a series of transfer products from a mixture of cyclomaltohexaose and cyclic tetrasaccharide (cyclo[-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->], CTS). Of the transfer products, only two components, saccharides A and D, remained and accumulated after digestion with glucoamylase. The total combined yield of the saccharides reached 63.4% of total sugars, and enzymatic and instrumental analyses revealed the structures of both saccharides. Saccharide A was identified as 4-mono-O-alpha-glucosyl-CTS, [-->6)-[alpha-D-Glcp-(1-->4)]-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->], and sachharide D was 4,4'-di-O-alpha-glucosyl-CTS, [-->6)-[alpha-D-Glcp-(1-->4)]-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-[alpha-D-Glcp-(1-->4)]-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->]. These structures led us to conclude that the glycosyltransfer catalyzed by CGTase was specific to the C4-OH of the 6-linked glucopyranosyl residues in CTS.
Shibuya
Takashi
T
Hayashibara Biochemical Laboratories, Inc., 7-7 Amase-minami machi, Okayama 700-0834, Japan. amaseken@hayashibara.co.jp
Aga
Hajime
H
Watanabe
Hikaru
H
Sonoda
Tomohiko
T
Kubota
Michio
M
Fukuda
Shigeharu
S
Kurimoto
Masashi
M
Tsujisaka
Yoshio
Y
D001234:Aspergillus niger / Q000201:enzymology; D001407:Bacillus / Q000201:enzymology; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry*; D006027:Glycosides / Q000737:chemistry*; D006031:Glycosylation; D006868:Hydrolysis; D013058:Mass Spectrometry; D008745:Methylation; D009844:Oligosaccharides / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D000520:alpha-Glucosidases / Q000378:metabolism; D001614:beta-Amylase / Q000378:metabolism
D006027:Glycosides; D009844:Oligosaccharides; D011134:Polysaccharides; C008315:maltodextrin; D005964:Glucosyltransferases; C019963:cyclomaltodextrin glucanotransferase; D001614:beta-Amylase; D000520:alpha-Glucosidases; D005087:Glucan 1,4-alpha-Glucosidase