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 Karaguler*

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13 Atıf (Scopus)

Özet

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.

Orijinal dilİngilizce
Sayfa (başlangıç-bitiş)507-520
Sayfa sayısı14
DergiExtremophiles
Hacim23
Basın numarası5
DOI'lar
Yayın durumuYayınlandı - 2 Eyl 2019

Bibliyografik not

Publisher Copyright:
© 2019, Springer Japan KK, part of Springer Nature.

Finansman

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

FinansörlerFinansör numarası
Istanbul Teknik Üniversitesi39574

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