title: Chemical synthesis and enzymatic, stereoselective hydrolysis of a functionalized dihydropyrimidine for the synthesis of β-amino acids.
abstract: A novel substrate, 6-(4-nitrophenyl)dihydropyrimidine-2,4(1H,3H)-dione (pNO2PheDU), was chemically synthesized and analytically verified for the potential biocatalytic synthesis of enantiopure β-amino acids. The hydantoinase (EC 3.5.2.2) from Arthrobacter crystallopoietes DSM20117 was chosen to prove the enzymatic hydrolysis of this substrate, since previous investigations showed activities of this enzyme toward 6-monosubstituted dihydrouracils. Whole cell biotransformations with recombinant Escherichia coli expressing the hydantoinase showed degradation of pNO2PheDU. Additionally, the corresponding N-carbamoyl-β-amino acid (NCarbpNO2 βPhe) was chemically synthesized, an HPLC-method with chiral stationary phases for detection of this product was established and thus (S)-enantioselectivity toward pNO2PheDU has been shown. Consequently this novel substrate is a potential precursor for the enantiopure β-amino acid para-nitro-β-phenylalanine (pNO2 βPhe).
authors.affiliation: Section II: Technical Biology, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany. christin.slomka@kit.edu.
authors.forename: Christin
authors.initials: C
authors.lastname: Slomka
authors.affiliation: Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, 76131, Karlsruhe, Germany. Sabilla.Zhong@Beiersdorf.com.
authors.forename: Sabilla
authors.initials: S
authors.lastname: Zhong
authors.affiliation: Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, 76131, Karlsruhe, Germany. anna.fellinger@gmx.net.
authors.forename: Anna
authors.initials: A
authors.lastname: Fellinger
authors.affiliation: Section II: Technical Biology, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany. ulrike.engel@kit.edu.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Engel
authors.affiliation: Section II: Technical Biology, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany. christoph.syldatk@kit.edu.
authors.forename: Christoph
authors.initials: C
authors.lastname: Syldatk
authors.affiliation: Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, 76131, Karlsruhe, Germany. stefan.braese@kit.edu.|Institute of Toxicology and Genetics, Institute of Technology (KIT), Hermann-von Helmholtz-Platz 1, 76344, Eggenstein-Leopoldshafen, Germany. stefan.braese@kit.edu.
authors.forename: Stefan
authors.initials: S
authors.lastname: Bräse
authors.affiliation: Section II: Technical Biology, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany. jens.rudat@kit.edu.
authors.forename: Jens
authors.initials: J
authors.lastname: Rudat
keywords: Dihydrouracil; Enantioselectivity; Enzyme catalysis; Hydrolases; β-Amino acids