Chemical modification of bacterial α-amylases:: changes in tertiary structures and the effect of additional calcium

A comparative study was performed on the effect of calcium on native and chemically modified forms of mesophilic and thermophilic alpha -amylases. Circular dichroism (CD) and irreversible thermoinactivation studies were carried out in the absence and presence of 10 mM calcium. From the CD experiments, changes in the tertiary structure of these enzymes, brought about by modification, were concluded. Furthermore, these changes were found to be influenced by the presence of calcium. Sorbitol was very effective in affording protection against irreversible thermoinactivation of native and modified forms of the enzymes, both in the absence and presence of calcium. Results are discussed in terms of the usefulness of this new approach involving a combination of medium and chemical modification for protein stabilization and enhancement of catalytic potential. (C) 2001 Elsevier Science BN. All rights reserved.