Studies of the inhibition by malto-oligosaccharides of the cyclisation reaction catalysed by the cyclodextrin glycosyltransferase from Klebsiella pneumoniae M 5 al with glycogen.

The substrate qualities of malto-oligosaccharides for the disproportionation reaction catalysed by the cyclodextrin glycosyltransferase [(1----4)-alpha-D-glucan:[(1----4)-alpha-D-glucopyranosyl]transferase (cyclising) EC 2.4.1.19] from Klebsiella pneumoniae M 5 al have been re-investigated. Maltose failed to be homologised with measurable velocity. The initial rates of disproportionation and the affinities of the enzyme increased with the chain lengths of the substrates. Maltopentaose was the smallest saccharide which, by disproportionation, yielded longer chains being cyclised initially. D-Glucose did not affect the initial cyclisation from glycogen, but served as acceptor for the "chain-shortening" reaction. Maltose inhibited the initial cyclisation reaction in a linearly competitive manner. Maltotriose and maltotetraose inhibited the cyclisation reaction competitively, the inhibition kinetics pointing to the binding of two effector-molecules to the enzyme. Competitive inhibition was also found with malto-pentaose, -hexaose, and -heptaose. The degrees of inhibition increased from maltose to maltotetraose, and decreased with the larger saccharides; maltotriose and maltotetraose were the most effective inhibitors of the initial cyclisation. Some possibilities for the subsite-mechanisms are discussed.