A novel glucanotransferase from a Bacillus circulans strain that produces a cyclomaltopentaose cyclized by an alpha-1,6-linkage.
A novel glucanotransferase, involved in the synthesis of a cyclomaltopentaose cyclized by an alpha-1,6-linkage [ICG5; cyclo-{-->6)-alpha-D-Glcp-(1-->4)-alpha-D-Glcp-(1-->4)-alpha-D-Glcp-(1-->4)-alpha-D-Glcp-(1-->4)-alpha-D-Glcp-(1-->}], from starch, was purified to homogeneity from the culture supernatant of Bacillus circulans AM7. The pI was estimated to be 7.5. The molecular mass of the enzyme was estimated to be 184 kDa by gel filtration and 106 kDa by SDS-PAGE. These results suggest that the enzyme forms a dimer structure. It was most active at pH 4.5 to 8.0 at 50 degrees C, and stable from pH 4.5 to 9.0 at up to 35 degrees C. The addition of 1 mM Ca(2+) enhanced the thermal stability of the enzyme up to 40 degrees C. It acted on maltooligosaccharides that have degrees of polymerization of 3 or more, amylose, and soluble starch, to produce ICG5 by an intramolecular alpha-1,6-glycosyl transfer reaction. It also catalyzed the transfer of part of a linear oligosaccharide to another oligosaccharide by an intermolecular alpha-1,4-glycosyl transfer reaction. Thus the ICG5-forming enzyme was found to be a novel glucanotransferase. We propose isocyclomaltooligosaccharide glucanotransferase (IGTase) as the trivial name of this enzyme.
Watanabe
Hikaru
H
Glycoscience Institute, Research Center, Hayashibara Biochemical Laboratories, Inc., Okayama, Japan. h-watanabe@hayashibara.co.jp
Nishimoto
Tomoyuki
T
Mukai
Kazuhisa
K
Kubota
Michio
M
Chaen
Hiroto
H
Fukuda
Shigeharu
S
D000595:Amino Acid Sequence; D000688:Amylose / Q000737:chemistry; D001407:Bacillus / Q000201:enzymology*; D003500:Cyclization; D019281:Dimerization; D005936:Glucans / Q000737:chemistry; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D005936:Glucans; D009844:Oligosaccharides; C514763:cyclo(-6-glucopyranosyl-1-4-glucopyranosyl-1-4-glucopyranosyl-1-4-glucopyranosyl-1-4-glucopyranosyl-1); C031146:maltoheptaose; D000688:Amylose; D005964:Glucosyltransferases; C514764:isocyclomaltooligosaccharide glucanotransferase, Bacillus circulans