caa a22 4500 44581876X CHVBK 20180317145237.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s00792-010-0344-1 doi (NATIONALLICENCE)springer-10.1007/s00792-010-0344-1 Molecular cloning, over expression and characterization of thermoalkalophilic esterases isolated from Geobacillus sp [Elektronische Daten] [Hasan Tekedar, Gülşah Şanlı-Mohamed] 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 741bp for Est1 and Est2, which encoded 246 amino acids and ORF of Est3 was 729bp 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 27.5kDa. 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 1,100U/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. Springer, 2010 Esterase nationallicence Geobacillus sp nationallicence Thermophiles nationallicence Alkalophiles nationallicence Overexpression nationallicence Tekedar Hasan Department of Chemistry, Science Faculty, İzmir Institute of Technology, Gülbahçe, Urla, İzmir, Turkey aut Şanlı-Mohamed Gülşah Department of Chemistry, Science Faculty, İzmir Institute of Technology, Gülbahçe, Urla, İzmir, Turkey aut Extremophiles Springer Japan 15/2(2011-03-01), 203-211 1431-0651 15:2<203 2011 15 792 https://doi.org/10.1007/s00792-010-0344-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-010-0344-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tekedar Hasan Department of Chemistry, Science Faculty, İzmir Institute of Technology, Gülbahçe, Urla, İzmir, Turkey aut NATIONALLICENCE 700 1- Şanlı-Mohamed Gülşah Department of Chemistry, Science Faculty, İzmir Institute of Technology, Gülbahçe, Urla, İzmir, Turkey aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/2(2011-03-01), 203-211 1431-0651 15:2<203 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer