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A cold-adapted esterase from psychrotrophic Pseudoalteromas sp. strain 643A

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Abstract

A psychrotrophic bacterium producing a cold-adapted esterase upon growth at low temperatures was isolated from the alimentary tract of Antarctic krill Euphasia superba Dana, and classified as Pseudoalteromonas sp. strain 643A. A genomic DNA library of strain 643A was introduced into Escherichia coli TOP10F’, and screening on tributyrin-containing agar plates led to the isolation of esterase gene. The esterase gene (estA, 621 bp) encoded a protein (EstA) of 207 amino acid residues with molecular mass of 23,036 Da. Analysis of the amino acid sequence of EstA suggests that it is a member of the GDSL-lipolytic enzymes family. The purification and characterization of native EstA esterase were performed. The enzyme displayed 20–50% of maximum activity at 0–20°C. The optimal temperature for EstA was 35°C. EstA was stable between pH 9 and 11.5. The enzyme showed activity for esters of short- to medium-chain (C4 and C10) fatty acids, and exhibited no activity for long-chain fatty acid esters like that of palmitate and stearate. EstA was strongly inhibited by phenylmethylsulfonyl fluoride, 2–mercaptoethanol, dithiothreitol and glutathione. Addition of selected divalent ions e.g. Mg2+, Co2+ and Cu2+ led to the reduction of enzymatic activity and the enzyme was slightly activated (∼30%) by Ca2+ ions.

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Acknowledgments

This work was supported by the Polish State Committee for Scientific Research Grant 2 P04B 018 30 to M.T. and N302 039 31/3427 to J.K.

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Authors and Affiliations

  1. Departament of Microbiology, Gdańsk University of Technology, ul. Narutowicza 11/12, 80-952, Gdansk, Poland

    Hubert Cieśliński, Anna Długołęcka & Józef Kur

  2. Institute of Technical Biochemistry (ITB), Technical University of Łódź, Stefanowskiego 4/10, 90-924, Lodz, Poland

    Aneta M. Białkowska, Maurycy Daroch, Karolina L. Tkaczuk, Halina Kalinowska & Marianna Turkiewicz

  3. Laboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology, Trojdena 4, 02-109, Warsaw, Poland

    Karolina L. Tkaczuk

Authors
  1. Hubert Cieśliński
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  2. Aneta M. Białkowska
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  3. Anna Długołęcka
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  4. Maurycy Daroch
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  5. Karolina L. Tkaczuk
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  6. Halina Kalinowska
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  7. Józef Kur
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  8. Marianna Turkiewicz
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Corresponding authors

Correspondence to Józef Kur or Marianna Turkiewicz.

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Cieśliński Hubert and Białkowska Aneta contributed equally to this work.

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Cieśliński, H., Białkowska, A.M., Długołęcka, A. et al. A cold-adapted esterase from psychrotrophic Pseudoalteromas sp. strain 643A. Arch Microbiol 188, 27–36 (2007). https://doi.org/10.1007/s00203-007-0220-2

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  • DOI: https://doi.org/10.1007/s00203-007-0220-2

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