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Cloning and Characterization of Cold-Adapted α-Amylase from Antarctic Arthrobacter agilis

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Abstract

In this study, the gene encoding an α-amylase from a psychrophilic Arthrobacter agilis PAMC 27388 strain was cloned into a pET-28a(+) vector and heterologously expressed in Escherichia coli BL21(DE3). The recombinant α-amylase with a molecular mass of about 80 kDa was purified by using Ni2+-NTA affinity chromatography. This recombinant α-amylase exhibited optimal activity at pH 3.0 and 30 °C and was highly stable at varying temperatures (30–60 °C) and within the pH range of 4.0–8.0. Furthermore, α-amylase activity was enhanced in the presence of FeCl3 (1 mM) and β-mercaptoethanol (5 mM), while CoCl2 (1 mM), ammonium persulfate (5 mM), SDS (10 %), Triton X-100 (10 %), and urea (1 %) inhibited the enzymatic activity. Importantly, the presence of Ca2+ ions and phenylmethylsulfonyl fluoride (PMSF) did not affect enzymatic activity. Thin layer chromatography (TLC) analysis showed that recombinant A. agilis α-amylase hydrolyzed starch, maltotetraose, and maltotriose, producing maltose as the major end product. These results make recombinant A. agilis α-amylase an attractive potential candidate for industrial applications in the textile, paper, detergent, and pharmaceutical industries.

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Acknowledgments

This study was financially supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (no. NRF-2015R1A2A2A01004733), by Golden Seed Project (213004-04-4-SBA30), Ministry of Agriculture, Ministry of Oceans and Fisheries, and by Korea Polar Research Institute grant (PE14070).

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Authors and Affiliations

  1. Department of Biotechnology and Bioengineering, Interdisciplinary Program for Bioenergy and Biomaterials, Chonnam National University, Gwangju, 61186, South Korea

    Su-mi Kim & Jong-il Choi

  2. Korea Polar Research Institute, Incheon, 21990, South Korea

    Hyun Park

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  1. Su-mi Kim
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Correspondence to Jong-il Choi.

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Kim, Sm., Park, H. & Choi, Ji. Cloning and Characterization of Cold-Adapted α-Amylase from Antarctic Arthrobacter agilis . Appl Biochem Biotechnol 181, 1048–1059 (2017). https://doi.org/10.1007/s12010-016-2267-5

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