, Volume 12, Issue 3, pp 311–323 | Cite as

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

  • Donatella de Pascale
  • Angela M. Cusano
  • Flavia Autore
  • Ermenegilda Parrilli
  • Guido di Prisco
  • Gennaro Marino
  • M. Luisa Tutino
Original Paper


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.


Pseudoalteromonashaloplanktis TAC125 Psychrophilic bacterial strain α/β Hydrolase 

Supplementary material

792_2008_163_MOESM1_ESM.tif (83 kb)
TABLE S1: Homologous sequences used as data set in phylogenetic analysis (TIF 82 kb)


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Copyright information

© Springer 2008

Authors and Affiliations

  • Donatella de Pascale
    • 1
  • Angela M. Cusano
    • 2
  • Flavia Autore
    • 2
  • Ermenegilda Parrilli
    • 2
    • 3
  • Guido di Prisco
    • 1
  • Gennaro Marino
    • 2
    • 3
  • M. Luisa Tutino
    • 2
    • 3
  1. 1.Institute of Protein Biochemistry, CNRNaplesItaly
  2. 2.Department of Organic Chemistry and BiochemistryUniversity of Naples Federico II, Complesso Universitario, Monte Sant’AngeloNaplesItaly
  3. 3.School of Biotechnological SciencesUniversity of Naples Federico II, Complesso Universitario Monte Sant’AngeloNaplesItaly

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