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Characterization of a recombinant amylolytic enzyme of hyperthermophilic archaeon Thermofilum pendens with extremely thermostable maltogenic amylase activity

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Abstract

A gene (Tpen_1458) encoding a putative alpha amylase from hyperthermophilic archaeon Thermofilum pendens (TfMA) was cloned and expressed in Escherichia coli. The recombinant amylolytic enzyme was purified by Ni-NTA affinity chromatography and its catalytic properties were examined. Purified TfMA was extremely thermostable with a half-life of 60 min at an optimal temperature of 95°C. TfMA activity increased to 136% in the presence of 5 mM CaCl2. Maximal activity was measured toward γ-cyclodextrin with a specific activity of 56 U/mg using copper bicinchoninate method. TfMA catalyzed the ring-opening reaction by cleaving one α-1,4-glycosidic linkage of cyclodextrin to produce corresponding single maltooligosaccharide at the initial time. The final products from cyclodextrins, linear maltooligosaccharides, and starch were glucose and maltose, and TfMA could also degrade pullulan and amylase inhibitor acarbose to panose and acarviosine-glucose, respectively. These results revealed that TfMA is a novel maltogenic amylase.

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Acknowledgments

This project was sponsored in part by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and a Research Grant from Jilin Province Government (20080253).

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Authors and Affiliations

  1. Key Laboratory of Agricultural Products Processing in Jilin Province Universities, Changchun University, Changchun, 130022, People’s Republic of China

    Xiaolei Li & Dan Li

  2. College of Biological and Agricultural Engineering, Jilin University, Changchun, 130022, People’s Republic of China

    Xiaolei Li & Yongguang Yin

  3. Center for Agricultural Biomaterials, and Department of Food Science and Biotechnology, School of Agricultural Biotechnology, Seoul National University, Seoul, 151-742, South Korea

    Kwan-Hwa Park

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  1. Xiaolei Li
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  2. Dan Li
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  3. Yongguang Yin
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  4. Kwan-Hwa Park
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Correspondence to Dan Li.

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Li, X., Li, D., Yin, Y. et al. Characterization of a recombinant amylolytic enzyme of hyperthermophilic archaeon Thermofilum pendens with extremely thermostable maltogenic amylase activity. Appl Microbiol Biotechnol 85, 1821–1830 (2010). https://doi.org/10.1007/s00253-009-2190-6

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  • DOI: https://doi.org/10.1007/s00253-009-2190-6

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