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

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Tarih

2019

Dergi Başlığı

Dergi ISSN

Cilt Başlığı

Yayıncı

Springer Japan KK

Erişim Hakkı

info:eu-repo/semantics/closedAccess

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 k(cat) value of 12.30 s(-1). The optimum pH and temperature of the enzyme were found as 9 and 30 degrees 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 Acigol 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.

Açıklama

Anahtar Kelimeler

Degenerate Pcr, Esterase, Genome Walking, Halophiles, Lake Acigol, Metagenomics

Kaynak

Extremophiles

WoS Q Değeri

Q3

Scopus Q Değeri

Q2

Cilt

23

Sayı

5

Künye