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Thermostable Esterase estUT1 from Ureibacillus thermosphaericus: Effect of TrxA Tag on the Enzyme Properties

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Abstract

The thermostable esterase from the bacterium Ureibacillus thermosphaericus was expressed with Trx tag from plasmid pET32b-estUT1 under T7 promoter in E. coli BL21(DE3). The specific activity and relative thermal stability of the tagged enzyme increased from 45.2 to 65.8% (1 h at 70°С). The additional TrxA tag does not affect the pH optimum of enzyme activity and substrate specificity. At the same time, the absence of the TrxA tag resulted in a significant increase in the stability of estUT1 in during incubation with various chemicals, including ethanol and methanol. The maximum catalytic efficiency (kcat/KM) for esterase was observed in the absence of the TrxA tag and was 280.0 s−1 mM−1. Thereby fusion with TrxA tag promotes the enzyme secretion in the dissolved form, but reduces its thermal stability.

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Funding

This work is part of a State task for the Boreskov Institute of Catalysis, project no. AAAA-A17-117041710075-0 “Scientific Bases of the Catalytic and Biotechnological Processes of Conversion of Renewable Raw Materials for Chemistry and Energy.”

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Authors and Affiliations

  1. Boreskov Institute of Catalysis, Siberian Branch, Russian Academy of Sciences, 630090, Novosibirsk, Russia

    K. N. Sorokina, Yu. V. Samoylova & V. N. Parmon

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  1. K. N. Sorokina
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  2. Yu. V. Samoylova
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  3. V. N. Parmon
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Correspondence to K. N. Sorokina, Yu. V. Samoylova or V. N. Parmon.

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Translated by G. Levit

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Sorokina, K.N., Samoylova, Y.V. & Parmon, V.N. Thermostable Esterase estUT1 from Ureibacillus thermosphaericus: Effect of TrxA Tag on the Enzyme Properties. Catal. Ind. 12, 148–154 (2020). https://doi.org/10.1134/S2070050420020099

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  • DOI: https://doi.org/10.1134/S2070050420020099

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