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Applied Microbiology and Biotechnology

, Volume 61, Issue 5–6, pp 517–522 | Cite as

Cloning and sequence analysis of the ces10 gene encoding a Sphingomonas paucimobilis esterase

  • P. A. Videira
  • A. M. Fialho
  • A. R. Marques
  • P. M. Coutinho
  • I. Sá-CorreiaEmail author
Original Paper

Abstract

The ces10 gene of the gellan gum-producing strain Sphingomonas paucimobilis ATCC 31461 was cloned and sequenced. Multi-sequence alignment of the deduced protein indicated that Ces10 belongs to the serine hydrolase family with a potential catalytic triad comprising Ser153 (within the G-X-S-X-G consensus sequence), His75 and Asp125. The mixed block results obtained following pattern search and the low identities detected in a BLAST analysis indicate that Ces10 is significantly different from other characterised bacterial esterases/lipases. Nevertheless, the Ces10 amino acid sequence showed 45% similarity with Rhodococcus sp. heroin esterase and 48% with Bacillus subtilis p-nitrobenzyl esterase. Ces10, with a predicted molecular mass of 30,641 Da, was overproduced in Escherichia coli and purified to homogeneity in a histidine-tagged form. Enzyme assays using p-nitrophenyl-esters (p-NP-esters) with different acyl chain-lengths as the substrate confirmed the anticipated esterase activity. Ces10 exhibited a marked preference for short-chain fatty acids, yielding the highest activity with p-NP-propionate (optimal pH 7.4, optimal temperature 37 °C).

Keywords

Lipolytic Enzyme Triacetine Ces10 Gene Sphingomonas Paucimobilis Ces10 Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was supported by the Fundação para a Ciência e Tecnologia, Portugal (project POCTI/35733/1999) and by post-doctoral and PhD grants to P.A.V. (SFRH/BPD/5710/2001) and A.R.M. (PRAXIS XXI/BD/18146/98)].

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • P. A. Videira
    • 1
  • A. M. Fialho
    • 1
  • A. R. Marques
    • 1
  • P. M. Coutinho
    • 1
  • I. Sá-Correia
    • 1
    Email author
  1. 1.Centro de Engenharia Biológica e QuímicaInstituto Superior TécnicoLisboaPortugal

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