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Characterization of the trehalosyl dextrin-forming enzyme from the thermophilic archaeon Sulfolobus solfataricus ATCC 35092

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Abstract

The trehalosyl dextrin-forming enzyme (TDFE) mainly catalyzes an intramolecular transglycosyl reaction to form trehalosyl dextrins from dextrins by converting the α-1,4-glucosidic linkage at the reducing end to an α-1,1-glucosidic linkage. In this study, the treY gene encoding TDFE was PCR cloned from the genomic DNA of Sulfolobus solfataricus ATCC 35092 to an expression vector with a T7 lac promoter and then expressed in Escherichia coli. The recombinant TDFE was purified sequentially by using heat treatment, ultrafiltration, and gel filtration. The obtained recombinant TDFE showed an apparent optimal pH of 5 and an optimal temperature of 75°C. The enzyme was stable in a pH range of 4.5–11, and the activity remained unchanged after a 2-h incubation at 80°C. The transglycosylation activity of TDFE was higher when using maltoheptaose as substrate than maltooligosaccharides with a low degree of polymerization (DP). However, the hydrolysis activity of TDFE became stronger when low DP maltooligosaccharides, such as maltotriose, were used as substrate. The ratios of hydrolysis activity to transglycosylation activity were in the range of 0.2–14% and increased when the DP of substrate decreased. The recombinant TDFE was found to exhibit different substrate specificity, such as its preferred substrates for the transglycosylation reaction and the ratio of hydrolysis to transglycosylation of the enzyme reacting with maltotriose, when compared with other natural or recombinant TDFEs from Sulfolobus.

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Acknowledgements

This work was supported by grant NSC 91-2313-B-019-039 from the National Science Council at Taiwan.

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Authors and Affiliations

  1. Department of Food Science, National Taiwan Ocean University, 2 Pei-Ning Rd., Keelung, 202, Taiwan

    Tsuei-Yun Fang, Xing-Guang Hung & Tong-Yuan Shih

  2. Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan

    Wen-Chi Tseng

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  1. Tsuei-Yun Fang
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  2. Xing-Guang Hung
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  3. Tong-Yuan Shih
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  4. Wen-Chi Tseng
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Correspondence to Tsuei-Yun Fang.

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Communicated by J. Wiegel

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Fang, TY., Hung, XG., Shih, TY. et al. Characterization of the trehalosyl dextrin-forming enzyme from the thermophilic archaeon Sulfolobus solfataricus ATCC 35092. Extremophiles 8, 335–343 (2004). https://doi.org/10.1007/s00792-004-0393-4

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