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Extremophiles

, Volume 23, Issue 5, pp 507–520 | Cite as

Recombinant production and characterization of a novel esterase from a hypersaline lake, Acıgöl, by metagenomic approach

  • Havva Esra Tutuncu
  • Nurgul Balci
  • Melek Tuter
  • Nevin Gul KaragulerEmail author
Original Paper

Abstract

The aim of this study was to isolate a novel esterase from a hypersaline lake by sequence-based metagenomics. The metagenomic DNA was isolated from the enriched hypersaline lake sediment. Degenerate primers targeting the conserved regions of lipolytic enzymes of halophilic microorganisms were used for polymerase chain reaction (PCR) and a whole gene was identified by genome walking. The gene was composed of 783 bp, which corresponds to 260 amino acids with a molecular weight of 28.2 kDa. The deduced amino acid sequence best matched with the esterase from Halomonas gudaonensis with an identity of 91%. Recombinantly expressed enzyme exhibited maximum activity towards pNP-hexanoate with a kcat value of 12.30 s−1. The optimum pH and temperature of the enzyme were found as 9 and 30 °C, respectively. The effects of NaCl, solvents, metal ions, detergents and enzyme inhibitors were also studied. In conclusion, a novel enzyme, named as hypersaline lake “Acıgöl” esterase (hAGEst), was identified by sequence-based metagenomics. The high expression level, the ability to maintain activity at cold temperatures and tolerance to DMSO and metal ions are the most outstanding properties of the hAGEst.

Keywords

Degenerate PCR Esterase Genome walking Halophiles Lake Acigöl Metagenomics 

Notes

Funding

This study was funded by Istanbul Technical University (project no: 39574).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

792_2019_1103_MOESM1_ESM.pdf (972 kb)
Supplementary material 1 (PDF 971 kb)

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Copyright information

© Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Havva Esra Tutuncu
    • 1
    • 2
    • 3
  • Nurgul Balci
    • 4
  • Melek Tuter
    • 5
  • Nevin Gul Karaguler
    • 1
    • 2
    Email author
  1. 1.Department of Molecular Biology and Genetics, Faculty of Science and LettersIstanbul Technical UniversityIstanbulTurkey
  2. 2.Istanbul Technical University Dr. Orhan Öcalgiray Molecular Biology-Biotechnology and Genetics Research CenterIstanbulTurkey
  3. 3.Department of Gastronomy and Culinary ArtsIstanbul Gedik UniversityIstanbulTurkey
  4. 4.Department of Geological Engineering, Faculty of MinesIstanbul Technical UniversityIstanbulTurkey
  5. 5.Department of Chemical Engineering, Faculty of Chemical and Metallurgical EngineeringIstanbul Technical UniversityIstanbulTurkey

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