Optimization of recombinant novel esterase expression from extremophiles

Yıl 2018, Cilt: 12 Sayı: 3, 33 - 36, 26.12.2018

Havva Esra Tütüncü , Nurgul Çelik Balci , Melek Tuter , Nevin Gul Karaguler

Öz

Esterases, which
are a sub-group of lipolytic enzymes, are important biocatalysts for many
industrial applications. In this study, optimization for the recombinant
expression of a novel esterase, which was previously obtained by metagenomic
approach, was studied. To optimize the expression, 0.1, 0.5 and 1 mM isopropyl
β-D-1 thiogalactopyranoside (IPTG) concentrations were tried. In addition,
induction at 25ºC for 16 hours, 30ºC for 6 hours and 37ºC for 3 hours were
tried. According to the results, induction at 30°C for 6 hours by 0.1 mM IPTG yielded
high amount of protein with maximum catalytic activity. After the gene was
successfully expressed, purification studies were conducted. The protein was
purified using His-tag method. Native and SDS-PAGE analysis showed that protein
which is present as a monomer was successfully purified. 

Anahtar Kelimeler

Protein expression, esterase

Kaynakça

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