%0 Journal Article %1 ISI:000078681800027 %A Siemann, M %A Alvarado-Marin, A %A Pietzsch, M %A Syldatk, C %C PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS %D 1999 %I ELSEVIER SCIENCE BV %J JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC %K acid} amidase; amino cyclic dihydropyrimidinase; hydantoin; myown protein purification; urease; {hydantoinase; %N 3 %P 387-397 %R 10.1016/S1381-1177(98)00137-4 %T A D-specific hydantoin amidohydrolase: properties of the metalloenzyme purified from Arthrobacter crystallopoietes %U https://doi.org/10.1016/S1381-1177(98)00137-4 %V 6 %X A D-specific hydantoinase has been purified to homogeneity from Arthrobacter crystallopoietes DSM 20117 with a yield of 5\% related to the crude extract. The active enzyme is a tetramer of 257 kDa consisting of four identical subunits, each with a molecular mass of 60 kDa. Incubation of the enzyme with the metal-chelating agent EDTA had no inhibitory effect, while 8-hydroxyquinoline-5-sulfonic acid resulted in a complete and irreversible inactivation. The purified enzyme contains zinc as cofactor, which could be detected by subjection to direct analysis using inductive/coupled plasma-atomic emission spectrometry. The hydantoinase has a wide substrate specificity for the D-selective cleavage of 5-monosubstituted hydantoin derivatives with aliphatic and aromatic side chains. The V-max-value for phenylhydantoin is 217 U/mg, the K-m-value is 8 mM. Dihydrouracil was found to be a natural substrate (V-max = 35 U/mg). The N-terminal amino acid sequence of the enzyme shows distinct homologies to other metal-dependent cyclic amidases involved in the nucleotide metabolism especially to dihydropyrimidinases as well as to ureases, L- and unselective hydantoinases. Due to these findings, this enzyme has to be considered as a possible link in the evolution to related L-selective and unselective hydantoinases from the genus of Arthrobacter. (C) 1999 Elsevier Science B.V. All rights reserved.

@article{ISI:000078681800027, abstract = {{A D-specific hydantoinase has been purified to homogeneity from Arthrobacter crystallopoietes DSM 20117 with a yield of 5\% related to the crude extract. The active enzyme is a tetramer of 257 kDa consisting of four identical subunits, each with a molecular mass of 60 kDa. Incubation of the enzyme with the metal-chelating agent EDTA had no inhibitory effect, while 8-hydroxyquinoline-5-sulfonic acid resulted in a complete and irreversible inactivation. The purified enzyme contains zinc as cofactor, which could be detected by subjection to direct analysis using inductive/coupled plasma-atomic emission spectrometry. The hydantoinase has a wide substrate specificity for the D-selective cleavage of 5-monosubstituted hydantoin derivatives with aliphatic and aromatic side chains. The V-max-value for phenylhydantoin is 217 U/mg, the K-m-value is 8 mM. Dihydrouracil was found to be a natural substrate (V-max = 35 U/mg). The N-terminal amino acid sequence of the enzyme shows distinct homologies to other metal-dependent cyclic amidases involved in the nucleotide metabolism especially to dihydropyrimidinases as well as to ureases, L- and unselective hydantoinases. Due to these findings, this enzyme has to be considered as a possible link in the evolution to related L-selective and unselective hydantoinases from the genus of Arthrobacter. (C) 1999 Elsevier Science B.V. All rights reserved.}}, added-at = {2018-01-25T13:38:08.000+0100}, address = {{PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}}, affiliation = {{Syldatk, C (Reprint Author), Univ Stuttgart, Inst Bioverfahrenstech, Allmandring 31, D-70569 Stuttgart, Germany. Univ Stuttgart, Inst Bioverfahrenstech, D-70569 Stuttgart, Germany.}}, author = {Siemann, M and Alvarado-Marin, A and Pietzsch, M and Syldatk, C}, author-email = {{syldatk@ivbt.uni-stuttgart.de}}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/212d2d3a837c7779a4133681896cdda2c/siemannherzberg}, da = {{2018-01-25}}, doc-delivery-number = {{168FV}}, doi = {{10.1016/S1381-1177(98)00137-4}}, interhash = {92a045b74da1fbef8200c77de7de663f}, intrahash = {12d2d3a837c7779a4133681896cdda2c}, issn = {{1381-1177}}, journal = {{JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC}}, journal-iso = {{J. Mol. Catal. B-Enzym.}}, keywords = {acid} amidase; amino cyclic dihydropyrimidinase; hydantoin; myown protein purification; urease; {hydantoinase;}, keywords-plus = {{BACILLUS-STEAROTHERMOPHILUS NS1122A; AMINO-ACIDS; MICROBIAL TRANSFORMATION; PURIFICATION; ENZYME; CRYSTALLIZATION; MICROORGANISMS}}, language = {{English}}, month = {{MAR 11}}, number = {{3}}, number-of-cited-references = {{37}}, orcid-numbers = {{Pietzsch, Markus/0000-0001-6266-9621}}, pages = {{387-397}}, publisher = {{ELSEVIER SCIENCE BV}}, research-areas = {{Biochemistry \& Molecular Biology; Chemistry}}, researcherid-numbers = {{Pietzsch, Markus/E-2026-2017}}, times-cited = {{12}}, timestamp = {2018-01-25T12:38:18.000+0100}, title = {{A D-specific hydantoin amidohydrolase: properties of the metalloenzyme purified from Arthrobacter crystallopoietes}}, type = {{Article}}, unique-id = {{ISI:000078681800027}}, url = {https://doi.org/10.1016/S1381-1177(98)00137-4}, usage-count-last-180-days = {{0}}, usage-count-since-2013 = {{0}}, volume = {{6}}, web-of-science-categories = {{Biochemistry \& Molecular Biology; Chemistry, Physical}}, year = {{1999}} }