caa a22 4500 605500789 CHVBK 20210128100557.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6803-y doi (NATIONALLICENCE)springer-10.1007/s00253-015-6803-y Expanding the threonine aldolase toolbox for the asymmetric synthesis of tertiary α-amino acids [Elektronische Daten] [Kateryna Fesko, Gernot Strohmeier, Rolf Breinbauer] The direct biochemical synthesis of tertiary α-amino acids with a wide range of diversity was recently reported using natural threonine aldolases LTA from Aeromonas jandei and DTA from Pseudomonas sp. Here, we describe the identification of five novel threonine aldolases which accept alanine and serine as amino acid donors. The enzymes were found by sequence database analysis using known aldolases as template. All enzymes were overexpressed in Escherichia coli and purified, and their biochemical properties were characterized. The new enantiocomplementary l- and d-threonine aldolases catalyze the asymmetric synthesis of β-hydroxy α-methyl- and α-hydroxymethyl-α-amino acids with good conversion and perfect enantioselectivity at α-carbon of the products (e.e. >99%). The structural basis for the broad donor specificity of these threonine aldolases is analyzed based on crystal structure alignments and amino acid sequences comparison. Springer-Verlag Berlin Heidelberg, 2015 Threonine aldolase nationallicence Aldol reactions nationallicence Tertiary amino acids nationallicence Enzyme catalysis nationallicence Biocatalysis nationallicence Fesko Kateryna Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010, Graz, Austria aut Strohmeier Gernot Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010, Graz, Austria aut Breinbauer Rolf Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010, Graz, Austria aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9651-9661 0175-7598 99:22<9651 2015 99 253 https://doi.org/10.1007/s00253-015-6803-y 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/s00253-015-6803-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Fesko Kateryna Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010, Graz, Austria aut NATIONALLICENCE 700 1- Strohmeier Gernot Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010, Graz, Austria aut NATIONALLICENCE 700 1- Breinbauer Rolf Institute of Organic Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010, Graz, Austria aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9651-9661 0175-7598 99:22<9651 2015 99 253