INCREASED CHEMICAL STABILITY OF BACILLUS LICHENIFORMIS α-AMYLASE UPON ACETYLATION
DOI:
https://doi.org/10.24193/subbchem.2017.2.25Keywords:
Bacillus licheniformis -amylase, acetylation, chemical stability, urea denaturationAbstract
Acetylated derivative (47%) of Bacillus licheniformis a-amylase (BLA) was prepared using acetic anhydride and its molecular properties including chemical stability were studied with the help of CD spectroscopy, analytical gel filtration and enzymatic activity measurements. Acetylated BLA preparation was found homogeneous with respect to charge and size based on electrophoretic and chromatographic results. Expansion in the molecular size of the modified BLA was evident from the decrease in its elution volume on Superdex 200 column as well as Stokes radius determination. Near-UV CD spectra suggested significant change in the tertiary structure of the acetylated BLA, whereas secondary structures remained unaltered, as judged from the far-UV CD spectra. Acetylated BLA displayed greater chemical stability against urea denaturation as revealed by the increase in the mid-point (Cm) of the denaturation curve and significant retention of biological activity at different urea concentrations. These results indicated greater conformational stability of acetylated BLA in the presence of urea.
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