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Thermostable CITase from Thermoanaerobacter thermocopriae shows negative cooperativity

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Abstract

Objective

The biochemical properties of a putative thermostable cycloisomaltooligosaccharide (CI) glucanotransferase gene from Thermoanaerobacter thermocopriae were determined using a recombinant protein (TtCITase) expressed in Escherichia coli and purified to a single protein.

Results

The 171-kDa protein displayed maximum activity at pH 6.0, and enzyme activity was stable at pH 5.0–11.0. The optimal temperature was 60 °C in 1 h incubation, and thermal stability of the protein was 63% at 60 °C for 24 h. TtCITase produced CI-7 to CI-17, as well as CI-18, CI-19, and CI-20, which are relatively large CIs. Additionally, an unusual kinetic feature of TtCITase was its negative cooperative behavior in the dextran T2000 cleavage reaction.

Conclusions

Based on our results, TtCITase can be applied to produce relatively large CIs at high temperature.

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Acknowledgments

This research was carried out with support from the “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ0131262019)” Rural Development Administration, Republic of Korea and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2015R1D1A1A01058227).

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Author notes

  1. So-Jin Yang and Su-Jeong Choi contributed equally to this work.

Authors and Affiliations

  1. Department of Food Science and Technology, Chonnam National University, Gwangju, 61186, Republic of Korea

    So-Jin Yang, Su-Jeong Choi & Young-Min Kim

  2. Department of Agrofood Resources, National Institute of Agricultural Sciences, Wanju, 55365, Republic of Korea

    Bo-Ram Park

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  1. So-Jin Yang
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  2. Su-Jeong Choi
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  3. Bo-Ram Park
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Correspondence to Bo-Ram Park or Young-Min Kim.

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Yang, SJ., Choi, SJ., Park, BR. et al. Thermostable CITase from Thermoanaerobacter thermocopriae shows negative cooperativity. Biotechnol Lett 41, 625–632 (2019). https://doi.org/10.1007/s10529-019-02666-6

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  • DOI: https://doi.org/10.1007/s10529-019-02666-6

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