1980 Volume 88 Issue 1 Pages 131-138
An α-amylase [EC 3. 2. 1. 1] from Streptomyces praecox was purified and its characteristic action, the conversion of maltotriose (G3) to maltose (G2) without appreciable formation of glucose (G1), was investigated. Isoelectric focusing or the glycogen adsorption procedure was employed after chromatography. Isoelectric focusing showed that the enzyme preparation after chromatographic separation comprises three isozymes. The preparation from the glycogen adsorption procedure showed the highest specific activity of any preparation of this enzyme ever obtained. Product analysis with uniformly labeled G3 revealed that at a high concentration (18mM) of G3, much more G2 is produced than G1 (the product ratio G2/G1 is over 20), while at a lower concentration (10 μM) the reaction mixture was composed of nearly equimolar amounts of glucose and maltose. Based on the product analysis of reducing endlabeled G3 in addition to the above findings, the following conversion mechanism is proposed: Streptomyces α-amylase catalyzes transglycosylation to produce maltotetraose (G4) as a transient product which is immediately degraded into two molecules of G2 by a subsequent hydrolytic reaction, i.e., two molecules of G3 are converted into three molecules of maltose without appreciable formation of glucose.
