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The cold-active Lip1 lipase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 is a member of a new bacterial lipolytic enzyme family

Extremophiles volume 12, pages 311–323 (2008)Cite this article

Abstract

The genome of the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 was searched for the presence of genes encoding ester-hydrolysing enzymes. Amongst the others, the gene PSHAa0051 coding for a putative secreted esterase/lipase was selected. The psychrophilic gene was cloned, functionally over-expressed in P. haloplanktis TAC125, and the recombinant product (after named PhTAC125 Lip1) was purified. PhTAC125 Lip1 was found to be associated to the outer membrane and exhibited higher enzymatic activity towards synthetic substrates with long acyl chains. A structural model was constructed using the structure of carboxylesterase Est30 from Geobacillus stearothermophilus as template. The model covered the central part of the protein with the exceptions of PhTAC125 Lip1 N- and C-terminal regions, where the psychrophilic protein displays extra-domains. The constructed model showed a typical α/β-hydrolase fold, and confirmed the presence of a canonical catalytic triad consisting of Ser, Asp and His. The sequence analysis showed that PhTAC125 Lip1 is distantly related to other lipolytic enzymes, but closely related to other putative psychrophilic esterases/lipases. The aligned proteins share common features, such as: (1) a conserved new active-site pentapeptide motif (LGG(F/L/Y)STG); (2) the likely extra-cytoplasmic localization, (3) the absence of a typical calcium-binding pocket, and (4) the absence of a canonical lid. These observations strongly suggest that aligned proteins constitute a novel lipase family, typical of psychrophilic marine γ-proteobacteria, and PhTAC125 Lip1 could be considered the first characterised member of this family.

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Acknowledgments

This work was supported by grants of Ministero dell’Università e della Ricerca Scientifica (Progetti di Rilevante Interesse Nazionale 2006) and the National Programme of Antarctic Research 2004 (PNRA). Support the Regional Center of Competence (CRdC ATIBB, Regione Campania—Naples) is gratefully acknowledged.

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Affiliations

  1. Institute of Protein Biochemistry, CNR, Via Pietro Castellino 111, 80131, Naples, Italy

    Donatella de Pascale & Guido di Prisco

  2. Department of Organic Chemistry and Biochemistry, University of Naples Federico II, Complesso Universitario, Monte Sant’Angelo, Via Cinthia 4, 80126, Naples, Italy

    Angela M. Cusano, Flavia Autore, Ermenegilda Parrilli, Gennaro Marino & M. Luisa Tutino

  3. School of Biotechnological Sciences, University of Naples Federico II, Complesso Universitario Monte Sant’Angelo, Via Cinthia 4, 80126, Naples, Italy

    Ermenegilda Parrilli, Gennaro Marino & M. Luisa Tutino

Authors
  1. Donatella de Pascale
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  2. Angela M. Cusano
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  3. Flavia Autore
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  4. Ermenegilda Parrilli
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  5. Guido di Prisco
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  6. Gennaro Marino
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  7. M. Luisa Tutino
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Correspondence to Donatella de Pascale.

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D. de Pascale and A. M. Cusano equally contributed to the work.

Communicated by K. Horikoshi.

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TABLE S1: Homologous sequences used as data set in phylogenetic analysis (TIF 82 kb)

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de Pascale, D., Cusano, A.M., Autore, F. et al. The cold-active Lip1 lipase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 is a member of a new bacterial lipolytic enzyme family. Extremophiles 12, 311–323 (2008). https://doi.org/10.1007/s00792-008-0163-9

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Keywords

  • Pseudoalteromonashaloplanktis TAC125
  • Psychrophilic bacterial strain
  • α/β Hydrolase