Expression and characterization of a new esterase with GCSAG motif from a permafrost metagenomic library

Lada E. Petrovskaya, Ksenia A. Novototskaya-Vlasova, Elena V. Spirina, Ekaterina V. Durdenko, Galina Yu Lomakina, Maria G. Zavialova, Evgeny N. Nikolaev, Elizaveta M. Rivkina

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31 Citations (Scopus)


As a result of construction and screening of a metagenomic library prepared from a permafrost-derived microcosm, we have isolated a novel gene coding for a putative lipolytic enzyme that belongs to the hormone-sensitive lipase family. It encodes a polypeptide of 343 amino acid residues whose amino acid sequence displays maximum likelihood with uncharacterized proteins from Sphingomonas species. A putative catalytic serine residue of PMGL2 resides in a new variant of a recently discovered GTSAG sequence in which a Thr residue is replaced by a Cys residue (GCSAG). The recombinant PMGL2 was produced in Escherichia coli cells and purified by Ni-affinity chromatography. The resulting protein preferably utilizes short-chain p-nitrophenyl esters (C4 and C8) and therefore is an esterase. It possesses maximum activity at 45°C in slightly alkaline conditions and has limited thermostability at higher temperatures. Activity of PMGL2 is stimulated in the presence of 0.25-1.5 M NaCl indicating the good salt tolerance of the new enzyme. Mass spectrometric analysis demonstrated that N-terminal methionine in PMGL2 is processed and cysteine residues do not form a disulfide bond. The results of the study demonstrate the significance of the permafrost environment as a unique genetic reservoir and its potential for metagenomic exploration.

Original languageEnglish
JournalFEMS Microbiology Ecology
Issue number5
Publication statusPublished - 31 Mar 2016
Externally publishedYes


  • Esterase
  • GCSAG motif
  • HSL family
  • Metagenome
  • Microcosm
  • Permafrost


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