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Cloning, Expression, Purification, and Characterization of a Novel Esterase from Lactobacillus plantarum

Molecular Biotechnology volume 44, pages 242–249 (2010)Cite this article

Abstract

Lactobacillus plantarum is an important lactic acid bacterium, usually found as natural inhabitant of food, such as fermented vegetables and meat products. However, little information about lactic acid bacteria, especially concerning L. plantarum, as a source of useful enzymes has been reported. The aim of this study was to clone, express in Escherichia coli, purify, and characterize an esterase from L. plantarum ATCC 8014. The esterase gene (1014 bp) was amplified and cloned in pET14b expression vector to express a His6-tagged protein in E. coli. Recombinant L. plantarum esterase was purified by Ni-NTA resin, presenting an apparent molecular mass of about 38 kDa. It presented highest activity at pH 6.0 and 40°C. Also, it presented preference for p-nitrophenyl butyrate, but hydrolyzed more efficiently p-nitrophenyl acetate. Besides, this study shows, for the first time, CD data about secondary structure of an esterase from L. plantarum.

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Acknowledgments

This study was financially supported by CNPq processes 476285/2007-0 and 552508/2007-1, and Rede Proteoma de Santa Catarina (FAPESC/FINEP/MCT). FCAB was recipient of a fellowship from CAPES, Ministry of Education, Brazil. JBB was recipient of a fellowship from CNPq, Ministry of Science and Technology, Brazil.

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Affiliations

  1. Department of Food Science and Technology, Universidade Federal de Santa Catarina, Rod. Admar Gonzaga, 1346, 88034-001, Florianópolis, Brazil

    Fábio Cristiano Angonesi Brod & Ana Carolina Maisonnave Arisi

  2. Departmento de Bioquímica, Universidade Federal de Santa Catarina, 88040-900, Florianópolis, Brazil

    Javier Vernal, Jean Borges Bertoldo & Hernán Terenzi

Authors
  1. Fábio Cristiano Angonesi Brod
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  2. Javier Vernal
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  3. Jean Borges Bertoldo
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  4. Hernán Terenzi
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  5. Ana Carolina Maisonnave Arisi
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Correspondence to Ana Carolina Maisonnave Arisi.

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Brod, F.C.A., Vernal, J., Bertoldo, J.B. et al. Cloning, Expression, Purification, and Characterization of a Novel Esterase from Lactobacillus plantarum . Mol Biotechnol 44, 242–249 (2010). https://doi.org/10.1007/s12033-009-9232-2

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  • DOI: https://doi.org/10.1007/s12033-009-9232-2

Keywords

  • L. plantarum
  • Esterase
  • E. coli expression
  • Biochemical characterization
  • Circular dichroism