Transglycosylation of glycosyl residues to cyclic tetrasaccharide by Bacillus stearothermophilus cyclomaltodextrin glucanotransferase using cyclomaltodextrin as the glycosyl donor

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2003-05, Vol.67 (5), p.1094-1100
• Main Authors: Shibuya, T. (Hayashibara Biochemical Labs. Inc., Okayama (Japan)), Aga, H, Watanabe, H, Sonoda, T, Kubota, M, Fukuda, S, Kurimoto, M, Tsujisaka, Y
• Format: Article
• Language: English
• Subjects: alpha-Glucosidases - metabolism; Aspergillus niger - enzymology; Bacillus - enzymology; BACILLUS STEAROTHERMOPHILUS; beta-Amylase - metabolism; BIOCHEMICAL REACTIONS; Biological and medical sciences; Catalysis; Chromatography, High Pressure Liquid; cyclic tetrasaccharide; cyclomaltodextrin; cyclomaltodextrin glucanotransferase; DEXTRINS; ENZYME REACTORS; Fundamental and applied biological sciences. Psychology; Geobacillus stearothermophilus - enzymology; Glucan 1,4-alpha-Glucosidase - metabolism; Glucosyltransferases - chemistry; GLYCOSIDES; Glycosides - chemistry; Glycosylation; Hydrolysis; Mass Spectrometry; Methylation; Oligosaccharides - chemistry; POLYSACCHARIDES; Polysaccharides - chemistry; TRANSFERASES; transglycosylation
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.67.1094

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)-a-D-Glcp-(1 ->3)-a-D-Glcp-(1->6)-a-D-Glcp-(1->3)-a-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-a-glucosyl-CTS, (->6)-[a-D-Glcp-(1->4)]-a-D-Glcp-(1->3)-a-D-Glcp-(1->6)-a-D-Glcp-(1->3)-a-D-Glcp-(1->), and sachharide D was 4,4'-di-O-a-glucosyl-CTS, (->6)-[a-D-Glcp-(1->4)]-a-D-Glcp-(1->3)-a-D-Glcp-(1->6)-[a-D-Glcp-(1->4)]-a-D-Glcp-(1->3)-a-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.