Enzymatic syntheses of various nonreducing oligosaccharides from starch have been reported by many researchers. Nonreducing oligosaccharides are generally divided into two groups: linear and cyclic oligosaccharides. Trehalose (α D glucopyranosyl α D glucopyranoside) is a typical linear nonreducing oligosaccharide occurring in bacteria, yeasts, fungi, plants and invertebrates. Mass production of trehalose from starch has been developed using two bacterial enzymes, maltooligosyltrehalose synthase (EC 5.4.99.15) and maltooligosyltrehalose trehalohydrolase (EC 3.2.1.141), 1 3) and now this saccharide is used in the food, cosmetics and the pharmaceutical industries. Cyclomaltohexaose (α cyclodextrin), one of the most well known cyclic oligosaccharides, is produced from linear α 1,4 glucans by the intramolecular α 1,4 transglycosylation reaction of a cyclomaltodextrin glucanotransferase (EC 2.4.1.19).4) The cyclic oligosaccharide has a hydrophobic cavity in the center of the structure. Guest molecules of suitable size can enter the cavity, and the formation of the inclusion complex is used for stabilizing labile materials, 5) masking odors 6) and modifying viscosity. 7) Co te and co workers first reported that a cyclic tetrasaccharide consist-(1 }, was produced from a dextran like polysaccharide, alternan, by its degrading enzyme.8 10) The cyclic oligosaccharide was designated cycloalternan (CA). Recently, we discovered two novel enzymes, 6 α glucosyltransferase and 3 α isomaltosyltransferase, in Bacillus globisporus, and succeeded in the mass production of this saccharide from starch by the joint reaction of the two enzymes.11 13) Although the characteristics of CA has not yet been totally clarified, potential applications of the saccharide in food, cosmetics and medicines are anticipated because of its unique structure. Hence nonreducing oligosaccharides pro-
)-α-D-Glcp-(1 4)-α-D-Glcp-(1 6)-α-D-Glcp-(1 4)-α-D-Glcp-(1 }, was designated cyclic maltosyl-maltose (CMM).CMM was not hydrolyzed by various amylases, such as α-amylase, β-amylase, glucoamylase, isoamylase, pullulanase, maltogenic α-amylase and α-glucosidase, but hydrolyzed by isomalto-dextranase to give rise to isomaltose. A glycosyltransferase involved in the synthesis of CMM from starch was purified to homogeneity from the culture supernatant of A. globiformis M6. The enzyme acted on maltooligosaccharides that have degrees of polymerization more than 3, amylose, and soluble starch to produce CMM but failed to act on cyclomaltodextrins, pullulan and dextran. The CMM-forming enzyme catalyzed both intermolecular and intramolecular α-1,6-maltosyl transfer reaction and found to be a novel maltosyltransferase (6MT). To reveal the degradation pathway of CMM, we identified two enzymes, CMM hydrolase (CMMase) and α-glucosidase, as the responsible enzymes from the cell-free extract of the strain. CMMase hydrolyzed CMM to maltose via maltosyl-maltose as intermediates; however, it did not hydrolyze CMM to glucose, suggesting that it is a novel hydrolase that hydrolyzes...