Analysis of the action pattern of sequential α-amylases from B. stearothermophilus and B. amyloliquefaciens on highly concentrated soluble starch.

Hydrolysis of highly concentrated soluble starch (60%, w/w) was performed using sequential α-amylases from Bacillus stearothermophilus (T, 0.2%, w/w) and Bacillus amyloliquefaciens (B, 0.1%, w/w) to identify their possible action patterns. We found that T reduced the average molecular weight (Mw) of soluble starch from 52,827 Da to 31,914 Da and significantly affected its branched chain length. Compared with soluble starch, the chains with DP 6-12 and DP ≥ 13 in the T samples were diminished by 46% and 96%, respectively. This resulted in an attenuation in the proportions of exterior and inner chains, as well as low iodine binding capacity of the hydrolysates. In contrast, a slower decrease in the average Mw of soluble starch occurred after TB incubation, and the level of DP 6-12 further lowered, causing a gradual decline in the iodine binding capacity of the hydrolysates. Gathered data revealed an unusual action pattern of sequential α-amylase treatment at high substrate concentrations. Bacillus stearothermophilus α-amylase exhibited more pronounced endo-hydrolysis of amylopectin, whereas the attack of Bacillus amyloliquefaciens α-amylase on the exterior chains was enhanced in amylopectin residues. These findings suggest that the synergy of various α-amylases is an effective strategy to promote the dextrinization of highly concentrated starch and finely modify the molecular structure of starch.