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

*此作品的通信作者

研究成果: 雜誌貢獻期刊論文同行評審

28 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

  • 化學 (全部)
  • 農業與生物科學 (全部)

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