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Characteristics of a High Maltose-Forming, Acid-Stable, and Ca2+-Independent α-amylase of the Acidophilic Bacillus acidicola

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Abstract

The purified acidic α-amylase of Bacillus acidicola is a monomer of 66.0 kDa, optimally active at pH 4.0 and 60 °C. The enzyme is Ca2+ independent with T 1/2 for 18 min at 80 °C. The K m, V max, and catalytic efficiency (k cat/K m) of the enzyme are 1.6 mg mL−1, 23.8 μmol mg−1 min−1, and 981 μmol s−1, respectively. Among detergents, Tween 20, 40, and 80 stimulated enzyme activity, whereas sodium dodecyl sulfate and Triton X-100 inhibited even at low concentration. EGTA has not affected the activity, whereas EDTA β-mercaptoethanol, iodoacetic acid, and Dithiothreitol exhibited a slight inhibitory action. Phenylmethanesulfonyl fluoride, N-bromosuccinimide, and Hg2+ strongly inhibited enzyme activity. The experimental activation energy and temperature quotient are 50.12 kJ mol−1 and 1.37. When thermodynamic parameters (ΔH and ΔS) of the enzyme have been determined at different temperatures, ΔG is positive suggesting that the enzyme is thermostable. The enzyme hydrolyzes raw starches, and therefore, the enzyme finds application in raw starch saccharification at sub-gelatinization temperatures that saves energy needed for gelatinization of raw starch at 105 °C.

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Acknowledgments

The authors gratefully acknowledge financial assistance from the Department of Science and Technology (DST) and Council of Scientific and Industrial Research (CSIR), Government of India, New Delhi during the course of this investigation.

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  1. Department of Microbiology, University of Delhi, South Campus, Benito, Juarez Road, New Delhi, 110021, India

    Archana Sharma & T. Satyanarayana

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  1. Archana Sharma
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  2. T. Satyanarayana
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Correspondence to T. Satyanarayana.

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Sharma, A., Satyanarayana, T. Characteristics of a High Maltose-Forming, Acid-Stable, and Ca2+-Independent α-amylase of the Acidophilic Bacillus acidicola . Appl Biochem Biotechnol 171, 2053–2064 (2013). https://doi.org/10.1007/s12010-013-0501-y

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