caa a22 4500 605504431 CHVBK 20210128100616.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6038-3 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6038-3 The environment shapes microbial enzymes: five cold-active and salt-resistant carboxylesterases from marine metagenomes [Elektronische Daten] [Anatoli Tchigvintsev, Hai Tran, Ana Popovic, Filip Kovacic, Greg Brown, Robert Flick, Mahbod Hajighasemi, Olga Egorova, Joseph Somody, Dmitri Tchigvintsev, Anna Khusnutdinova, Tatyana Chernikova, Olga Golyshina, Michail Yakimov, Alexei Savchenko, Peter Golyshin, Karl-Erich Jaeger, Alexander Yakunin] Most of the Earth's biosphere is cold and is populated by cold-adapted microorganisms. To explore the natural enzyme diversity of these environments and identify new carboxylesterases, we have screened three marine metagenome gene libraries for esterase activity. The screens identified 23 unique active clones, from which five highly active esterases were selected for biochemical characterization. The purified metagenomic esterases exhibited high activity against α-naphthyl and p-nitrophenyl esters with different chain lengths. All five esterases retained high activity at 5°C indicating that they are cold-adapted enzymes. The activity of MGS0010 increased more than two times in the presence of up to 3.5M NaCl or KCl, whereas the other four metagenomic esterases were inhibited to various degrees by these salts. The purified enzymes showed different sensitivities to inhibition by solvents and detergents, and the activities of MGS0010, MGS0105 and MGS0109 were stimulated three to five times by the addition of glycerol. Screening of purified esterases against 89 monoester substrates revealed broad substrate profiles with a preference for different esters. The metagenomic esterases also hydrolyzed several polyester substrates including polylactic acid suggesting that they can be used for polyester depolymerization. Thus, esterases from marine metagenomes are cold-adapted enzymes exhibiting broad biochemical diversity reflecting the environmental conditions where they evolved. Springer-Verlag Berlin Heidelberg, 2014 Marine metagenome nationallicence Metagenomic library nationallicence Esterase nationallicence Cold adaptation nationallicence Salt resistance nationallicence Polyester hydrolysis nationallicence Tchigvintsev Anatoli Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Tran Hai Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Popovic Ana Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Kovacic Filip Institute of Molecular Enzyme Technology, Heinrich-Heine-University Duesseldorf, Forschungszentrum Juelich, 52426, Juelich, Germany aut Brown Greg Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Flick Robert Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Hajighasemi Mahbod Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Egorova Olga Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Somody Joseph Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Tchigvintsev Dmitri Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Khusnutdinova Anna Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Chernikova Tatyana School of Biological Sciences, Bangor University, LL57 2UW, Gwynedd, UK aut Golyshina Olga School of Biological Sciences, Bangor University, LL57 2UW, Gwynedd, UK aut Yakimov Michail Institute for Coastal Marine Environment, CNR, 98122, Messina, Italy aut Savchenko Alexei Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Golyshin Peter School of Biological Sciences, Bangor University, LL57 2UW, Gwynedd, UK aut Jaeger Karl-Erich Institute of Molecular Enzyme Technology, Heinrich-Heine-University Duesseldorf, Forschungszentrum Juelich, 52426, Juelich, Germany aut Yakunin Alexander Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2165-2178 0175-7598 99:5<2165 2015 99 253 https://doi.org/10.1007/s00253-014-6038-3 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6038-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tchigvintsev Anatoli Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Tran Hai Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Popovic Ana Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Kovacic Filip Institute of Molecular Enzyme Technology, Heinrich-Heine-University Duesseldorf, Forschungszentrum Juelich, 52426, Juelich, Germany aut NATIONALLICENCE 700 1- Brown Greg Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Flick Robert Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Hajighasemi Mahbod Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Egorova Olga Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Somody Joseph Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Tchigvintsev Dmitri Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Khusnutdinova Anna Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Chernikova Tatyana School of Biological Sciences, Bangor University, LL57 2UW, Gwynedd, UK aut NATIONALLICENCE 700 1- Golyshina Olga School of Biological Sciences, Bangor University, LL57 2UW, Gwynedd, UK aut NATIONALLICENCE 700 1- Yakimov Michail Institute for Coastal Marine Environment, CNR, 98122, Messina, Italy aut NATIONALLICENCE 700 1- Savchenko Alexei Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Golyshin Peter School of Biological Sciences, Bangor University, LL57 2UW, Gwynedd, UK aut NATIONALLICENCE 700 1- Jaeger Karl-Erich Institute of Molecular Enzyme Technology, Heinrich-Heine-University Duesseldorf, Forschungszentrum Juelich, 52426, Juelich, Germany aut NATIONALLICENCE 700 1- Yakunin Alexander Department of Chemical Engineering and Applied Chemistry, University of Toronto, M5S 3E5, Toronto, ON, Canada aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2165-2178 0175-7598 99:5<2165 2015 99 253