Identification and molecular modeling of a novel lipase from an Antarctic soil metagenomic library

• Autorzy: Cieslinski H , Bialkowska A. , Tkaczuk K. , Dlugolecka A. , Kur J. , Turkiewicz M.
• Języki publikacji: EN

Abstrakty

EN

In this work, we present the construction of a metagenomic library in Escherichia coli using pUC19 vector and environmental DNA directly isolated from Antarctic topsoil and screened for lipolytic enzymes. Screening on agar supplemented with olive oil and rhodamine B revealed one clone with lipolytic activity (Lip 1) out of 11000 E. coli clones. This clone harbored a plasmid, pLip 1, which has an insert of 4722 bp that was completely sequenced from both directions. Further analysis of the insert showed three open reading frames (ORFs). ORF2 encoded a protein (Lip 1) of 469 amino acids with 93% identity to the uncultured Pseudomonas sp. lipase LipJ03. Amino acid sequence comparison and phylogenetic analysis indicated that Lip 1 lipase was closely related to family I subfamily 3. Furthermore, we present a three-dimensional model of lipase Lip 1 which was generated based on the two known structures of mesophilic lipases from Pseudomonas sp. MIS 38 (PML lipase, PDB; 2Z8X) and Seiratia marcescens (SML lipase, PDB: 2QUB). Finally, we report the results of comparisons between lipase Lip 1 and mesophilic lipases and point out similarities and differences in the catalytic site and in other parts of the analyzed structures.

Słowa kluczowe

EN

genomic DNA; bacteria; Pseudomonas; lipase; lipolytic enzyme; Antarctic soil; identification; soil; Escherichia coli; Serratia marcescens; environmental sample; molecular modelling; metagenomics

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2009
• Tom: 58
• Numer: 3
• Opis fizyczny: p.199-204,fig.,ref.

Twórcy

autor

Cieslinski H

• Gdansk University of Technology, Narutowicza 11/12, 80-952 Gdansk, Poland

autor

Bialkowska A.

autor

Tkaczuk K.

autor

Dlugolecka A.

autor

Kur J.

autor

Turkiewicz M.

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