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Stereoselective hydrolysis of aryl-substituted dihydropyrimidines by hydantoinases

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Abstract

In this study, we investigated the possibility of using a modified hydantoinase process for the production of optically pure β-amino acids. Two aryl-substituted dihydropyrimidines d,l-6-phenyl-5,6-dihydrouracil (PheDU) and para-chloro-d,l-6-phenyl-5,6-dihydrouracil (pClPheDU) were synthesized. Hydrolysis of these novel substrates to the corresponding N-carbamoyl-β-amino acids by three recombinant d-hydantoinases and several bacterial strains was tested. All applied recombinant d-hydantoinases and eight bacterial isolates catalyzed the conversion of PheDU to N-carbamoyl-β-phenylalanine (NCβPhe). Some of these biocatalysts showed an enantioselectivity for either the d- or the l-PheDU enantiomer. The second dihydropyrimidinase substrate pClPheDU was hydrolyzed by all three recombinant d-hydantoinases and six of the wild-type strains. To our knowledge, this is the first dihydropyrimidinase activity reported with this aryl-substituted dihydropyrimidine. For selected biocatalysts, hydantoinase activity towards aryl-substituted hydantoins was demonstrated as well. However, none of the bacterial strains tested so far exhibited any carbamoylase activity towards NCβPhe.

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Acknowledgments

This work was financed by the Federal Ministry of Science and Education (BMBF), Germany. Furthermore, the authors like to thank Gerd Unkelbach from Fraunhofer Institute for Chemical Technology for cooperation in the synthesis of the above-mentioned substrates. We also thank Professor Don Cowan for providing the soil samples for the screening experiments.

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  1. Karlsruhe Institute of Technology (KIT), Institute of Process Engineering in Life Sciences, Section II: Technical Biology, Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany

    U. Engel, C. Syldatk & J. Rudat

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  1. U. Engel
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  2. C. Syldatk
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Correspondence to U. Engel.

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Engel, U., Syldatk, C. & Rudat, J. Stereoselective hydrolysis of aryl-substituted dihydropyrimidines by hydantoinases. Appl Microbiol Biotechnol 94, 1221–1231 (2012). https://doi.org/10.1007/s00253-011-3691-7

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