Role of the C-terminal domain of Thermus thermophilus trehalose synthase in the thermophilicity, thermostability, and efficient production of trehalose

Jia Hung Wang, Meng Yin Tsai, Jen Jye Chen, Guan Chiun Lee, Jei Fu Shaw

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    26 引文 斯高帕斯(Scopus)

    摘要

    Trehalose synthase (TS) from Thermus thermophilus (TtTS) is a thermostable enzyme that catalyzes the conversion of maltose into trehalose by intramolecular transglucosylation. It has a relatively higher thermophilicity and thermostability and a better conversion ratio for trehalose production than other known TSs from different sources at present. By amino acid sequences and the schematic motif alignment of trehalose synthase-related enzymes, it was found that TtTS (965 amino acid residues) contains a particular C-terminal fragment that is not found in most other TSs. To verify the function of this fragment, C-terminal deletion and enzyme fusion were respectively performed to explain the important role this fragment plays in the formation of trehalose. First, the C terminus (TtTSΔN, 415 amino acid residues) of TtTS is deleted to construct a TtTSΔC containing 550 amino acids. Furthermore, a novel cold-active TS was cloned and purified from Deinococcus radiodurans (DrTS, 552 amino acid residues) and then a fusion protein was created with TtTSΔN at the C terminus of DrTS (DrTS-TtTSΔN). It was found that the recombinant TtTSΔC enzyme had a lower thermostability and a higher byproduct than TtTS in catalyzing the conversion of maltose into trehalose. On the other hand, the recombinant DrTS-TtTSΔN enzyme had a higher thermostability and a lower byproduct than DrTS in their reactions. The above-mentioned results allowed the inference that the C terminus of TtTS plays a key role in maintaining its thermostability and hence in modulating the side reaction to reduce glucose production at a high temperature. A new, simple, and fast method to improve thermophilicity by fusing this fragment with particular conformation to a thermolabile enzyme is offered.

    原文英語
    頁(從 - 到)3435-3443
    頁數9
    期刊Journal of Agricultural and Food Chemistry
    55
    發行號9
    DOIs
    出版狀態已發佈 - 2007 五月 2

    ASJC Scopus subject areas

    • Chemistry(all)
    • Agricultural and Biological Sciences(all)

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