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Molecular cloning, characterization, and application of a novel thermostable α-glucosidase from the hyperthermophilic archaeon Pyrobaculum aerophilum strain IM2

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Abstract

A novel hyperthermostable α-glucosidase from the hyperthermophilic archaeon Pyrobaculum aerophilum str. IM2 (PAE1968) was successfully expressed in Escherichia coli and characterized. Recombinant PAE1968 was purified using Ni-NTA affinity chromatography to reveal a glycosidase with a predicted molecular weight (Mw) of 76 kDa. PAE1968 liberated glucose from the non-reducing ends of oligosaccharides. Multiple sequence alignment confirmed that the enzyme belonged to glycoside hydrolase (GH) family 31, and a kinetic study showed that the enzyme had a substrate preference for maltose (G2), indicating a Type-II α-glucosidase. The optimum operating conditions for the enzyme were 90°C and pH 6.0, while the enzyme retained at least 80% of maximal activity at pH 4.5–7.5. Transglycosylation and reversion reactions of PAE1968 were of a lower magnitude than for a commercial saccharification enzyme, indicating that this enzyme can be used for glucose production in the starch industry.

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Authors and Affiliations

  1. Department of Food Science and Nutrition, and Center for Aging and Health Care, Hallym University, Chuncheon, Gangwon, 200-702, Korea

    Hyeyeon Jeon, Hyewon Lee, Dahye Byun, Hyejeong Choi & Jae-Hoon Shim

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  1. Hyeyeon Jeon
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  2. Hyewon Lee
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  3. Dahye Byun
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  5. Jae-Hoon Shim
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Correspondence to Jae-Hoon Shim.

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Jeon, H., Lee, H., Byun, D. et al. Molecular cloning, characterization, and application of a novel thermostable α-glucosidase from the hyperthermophilic archaeon Pyrobaculum aerophilum strain IM2. Food Sci Biotechnol 24, 175–182 (2015). https://doi.org/10.1007/s10068-015-0024-0

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