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Two Novel Acetylesterases from Pantoea dispersa: Recombinant Expression, Purification, and Characterization

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Abstract

Two novel acetylesterases from Pantoea dispersa, with low amino acid sequence identity between them, were expressed in Escherichia coli with a carboxyl-His6 tail given by the expression plasmid, purified, and characterized. The purified proteins, named Est-1 and Est-2, had a molecular mass of 33 kDa and 37 kDa, respectively. Both proteins presented a modeled structure of homodimers with monomers presenting the α/β-hydrolase fold, with the catalytic triad Ser-Asp-His present in the active site. The KM for p-nitrophenyl acetate and Vmax values found for Est-1 were of 1.4 ± 0.2 mM and 8.66 ± 0.59 μmol/min and for Est-2 were of 0.36 ± 0.077 mM and 6.13 ± 0.56 μmol/min, respectively. Both enzymes presented an optimum pH of 7.0. The optimum temperature for Est-1 was 40 °C and for Est-2 was 50 °C. The temperatures in which the enzymes Est-1 and Est-2 lost half of their activity (T50) were 44.1 and 58.9 °C, respectively. SDS, EDTA, and PMSF significantly inhibited the enzymes. The two purified enzymes also presented activity against triacetin and were able to deacetylate the carbohydrates pectin and xylan, with higher activity against pectin. Thus, they could be considered as carbohydrate esterases.

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  • 03 July 2019

    The original version of this article unfortunately contained a mistake. Under Materials and Methods heading, Bacterial Strains sub-heading, the correct name of the used strain is “FEI4 65” and not “FzEI4 65.”

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Funding

The authors are thankful to the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes) and to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazil, for the Project funding (grant 001) and the scholarship given to D.B. Martim, respectively.

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  1. Department of Biochemistry, State University of Maringá, Av. Colombo, 5790, Maringá, PR, 87020-900, Brazil

    Damaris Batistão Martim & Ione Parra Barbosa-Tessmann

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The original version of this article was revised: The original version of this article unfortunately contained a mistake. Under Materials and Methods heading, Bacterial Strains sub-heading, the correct name of the used strain is “FEI4 65” and not “FzEI4 65.”

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Martim, D.B., Barbosa-Tessmann, I.P. Two Novel Acetylesterases from Pantoea dispersa: Recombinant Expression, Purification, and Characterization. Appl Biochem Biotechnol 189, 834–854 (2019). https://doi.org/10.1007/s12010-019-03024-y

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