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Molecular cloning and characterization of a thermostable α-amylase exhibiting an unusually high activity

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Abstract

An α-amylase gene was cloned from the thermophilic bacterium Bacillus subtilis isolated from Indonesian oil palm shell waste. The gene expressed an extracellular enzyme. Optimal hydrolysis conditions for the enzyme were 70°C and pH 6.0. The specific activity of the enzyme was 16.0 kU per mg of protein, which was higher than for other thermostable amylases. Hydrolytic products of the enzyme using starch and glycogen were mainly maltohexaose and maltopentaose. The enzyme had a K m value of 0.099 mg/mL for amylopectin, more than 10 times lower than for amylose. The catalytic efficiency of the enzyme using amylopectin was 39,200 mL/mg·s and was 3,270 mL/mg·s using amylose. The enzyme liquefied corn starch at pH 5.0, which was successfully converted to glucose using commercial glucoamylase and pullulanase without pH adjustment. The enzyme has advantages for industrial applications.

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Authors and Affiliations

  1. Department of Food Science and Human Nutrition, Iowa State University, Iowa, IA, 50011, USA

    Jong-Tae Park & Jay-lin Jane

  2. Department of Food Science and Technology, Chungnam National University, Daejeon, 305-764, Korea

    Jong-Tae Park

  3. Department of Biology, Faculty of Science and Mathematics, Bogor Agricultural University, Bogor, Kediri, 16680, Indonesia

    Antonius Suwanto

  4. Charoen Pokphand Indonesia, Jakarta Utara, 14430, Kediri, Indonesia

    Irawan Tan & Tommy Nuryanto

  5. Biotech Department of PT, BISI International, Kediri, 16720, Indonesia

    Rudy Lukman

  6. Department of Agronomy, Iowa State University, Ames, IA, 50011, USA

    Kan Wang

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  1. Jong-Tae Park
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  2. Antonius Suwanto
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  7. Jay-lin Jane
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Correspondence to Jay-lin Jane.

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Park, JT., Suwanto, A., Tan, I. et al. Molecular cloning and characterization of a thermostable α-amylase exhibiting an unusually high activity. Food Sci Biotechnol 23, 125–132 (2014). https://doi.org/10.1007/s10068-014-0017-4

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  • DOI: https://doi.org/10.1007/s10068-014-0017-4

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