title: A thermophilic-like ene-reductase originating from an acidophilic iron oxidizer.
abstract: Ene-reductases originating from extremophiles are gaining importance in the field of biocatalysis due to higher-stability properties. The genome of the acidophilic iron-oxidizing bacterium "Ferrovum" sp. JA12 was found to harbor a thermophilic-like ene-reductase (FOYE-1). The foye-1 gene was ligated into a pET16bp expression vector system, and the enzyme was produced in Escherichia coli BL21 (DE3; pLysS) cells in yields of 10 mg L-1. FOYE-1 showed remarkable activity and rates on N-phenylmaleimide and N-phenyl-2-methylmaleimide (up to 89 U mg-1, >97 % conversion, 95 % (R)-selective) with both nicotinamide cofactors, NADPH and NADH. The catalytic efficiency with NADPH was 27 times higher compared to NADH. At the temperature maximum (50 °C) and pH optimum (6.5), activity was almost doubled to 160 U mg-1. These findings accomplish FOYE-1 for a valuable biocatalyst in the synthesis of succinimides. The appearance of a thermophilic-like ene-reductase in an acidic habitat is discussed with respect to its phylogenetic placement and to the genomic neighborhood of the encoding gene, awarding FOYE-1 a putative involvement in a quorum-sensing process.
authors.affiliation: Interdisciplinary Ecological Center, Institute of Bioscience, Environmental Microbiology Group, Technical University Bergakademie Freiberg, 09599, Freiberg, Germany. anika.scholtissek@gmail.com.|Department of Biotechnology, Delft University of Technology, 2628BL, Delft, the Netherlands. anika.scholtissek@gmail.com.
authors.forename: Anika
authors.identifier: http://orcid.org/0000-0002-6506-1467
authors.initials: A
authors.lastname: Scholtissek
authors.affiliation: Interdisciplinary Ecological Center, Institute of Bioscience, Environmental Microbiology Group, Technical University Bergakademie Freiberg, 09599, Freiberg, Germany.
authors.forename: Sophie R
authors.initials: SR
authors.lastname: Ullrich
authors.affiliation: Interdisciplinary Ecological Center, Institute of Bioscience, Environmental Microbiology Group, Technical University Bergakademie Freiberg, 09599, Freiberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Mühling
authors.affiliation: Interdisciplinary Ecological Center, Institute of Bioscience, Environmental Microbiology Group, Technical University Bergakademie Freiberg, 09599, Freiberg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schlömann
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2628BL, Delft, the Netherlands.
authors.forename: Caroline E
authors.initials: CE
authors.lastname: Paul
authors.affiliation: Interdisciplinary Ecological Center, Institute of Bioscience, Environmental Microbiology Group, Technical University Bergakademie Freiberg, 09599, Freiberg, Germany. dirk-tischler@email.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Tischler
mesh_terms: D020562:Betaproteobacteria / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D003067:Coenzymes / Q000032:analysis; D019295:Computational Biology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D016680:Genome, Bacterial; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008301:Maleimides / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013388:Succinimides / Q000378:metabolism; D013696:Temperature
chemical_list: D003067:Coenzymes; D008301:Maleimides; D011994:Recombinant Proteins; D013388:Succinimides; C028525:N-phenylmaleimide; D010088:Oxidoreductases
keywords: Biocatalysis; Extremophil; Ferrovum; Flavoprotein; Old yellow enzyme