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