Please use this identifier to cite or link to this item: https://hdl.handle.net/11147/5099
Title: Molecular cloning, over expression and characterization of thermoalkalophilic esterases isolated from Geobacillus sp
Authors: Tekedar, Hasan Cihad
Şanlı Mohamed, Gülşah
Tekedar, Hasan Cihad
Şanlı Mohamed, Gülşah
Izmir Institute of Technology. Chemistry
Keywords: Alkalophiles
Esterase
Geobacillus sp.
Overexpression
Thermophiles
Issue Date: Mar-2011
Publisher: Springer Verlag
Source: Tekedar, H.C., and Şanlı Mohamed, G. (2011). Molecular cloning, over expression and characterization of thermoalkalophilic esterases isolated from Geobacillus sp. Extremophiles, 15(2), 203-211. doi:10.1007/s00792-010-0344-1
Abstract: Due to potential use for variety of biotechnological applications, genes encoding thermoalkalophilic esterase from three different Geobacillus strains isolated from thermal environmental samples in Balçova (Agamemnon) geothermal site were cloned and respective proteins were expressed in Escherichia coli (E. coli) and characterized in detail. Three esterases (Est1, Est2, Est3) were cloned directly by PCR amplification using consensus degenerate primers from genomic DNA of the strains Est1, Est2 and Est3 which were from mud, reinjection water and uncontrolled thermal leak, respectively. The genes contained an open reading frame (ORF) consisting of 741 bp for Est1 and Est2, which encoded 246 amino acids and ORF of Est3 was 729 bp encoded 242 amino acids. The esterase genes were expressed in E. coli and purified using His-Select HF nickel affinity gel. The molecular mass of the recombinant enzyme for each esterase was approximately 27. 5 kDa. The three esterases showed high specific activity toward short chain p-NP esters. Recombinant Est1, Est2, Est3 have exhibited similar activity and the highest esterase activity of 1,100 U/mg with p-nitrophenyl acetate (pNPC2) as substrate was observed with Est1. All three esterase were most active around 65°C and pH 9.5-10.0. The effect of organic solvents, several metal ions, inhibitors and detergents on enzyme activity for purified Est1, Est2, Est3 were determined separately and compared.
URI: https://doi.org/10.1007/s00792-010-0344-1
http://hdl.handle.net/11147/5099
ISSN: 1431-0651
1431-0651
Appears in Collections:Chemistry / Kimya
Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

Files in This Item:
File Description SizeFormat 
5099.pdfMakale823.63 kBAdobe PDFThumbnail
View/Open
Show full item record

CORE Recommender

SCOPUSTM   
Citations

14
checked on Sep 25, 2021

WEB OF SCIENCETM
Citations

16
checked on Sep 25, 2021

Page view(s)

24
checked on Sep 28, 2021

Download(s)

30
checked on Sep 28, 2021

Google ScholarTM

Check

Altmetric


Items in GCRIS Repository are protected by copyright, with all rights reserved, unless otherwise indicated.