Crystal Structure of Metal-Dependent Allantoinase from Escherichia coli

Structural and Functional Insights into (S)-Ureidoglycine Aminohydrolase, Key Enzyme of Purine Catabolism in Arabidopsis thaliana

“…The modifications were performed using the QuikChange method (Agilent) to introduce a silent mutation at the sequences for NdeI and XhoI sites residing in the internal region of the AtUGlyAH gene. The N-terminal truncated variants of AtUGlyAH, from which the signal peptide (residues [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] and an additional N-terminal region had been removed, were amplified using a pair of sequence-specific primers (supplemental Table S1). The resulting PCR products were introduced into the NdeI and XhoI sites of a modified pET-28a vector (Merck) containing a tobacco etch virus protease cleavage site at the junction between a His 5 tag and a multiple cloning site.…”

“…Therefore, expression vectors were constructed as described above for genes encoding the enzymes necessary for producing (S)-ureidoglycine from a racemic allantoin mixture and also for the gene encoding (S)-ureidoglycolate dehydrogenase. Those were as follows: allantoinase from E. coli (14), allantoate amidohydrolase from A. thaliana (gene access no. NM_11826) and E. coli strain K-12 substrain DH10b (EcAAH; gene access no.…”

“…Subsequently, the N-terminal His-tagged protein was purified by immobilized metal affinity chromatography and dialyzed against buffer (50 mM Tris-HCl, pH 7.4, and 200 mM NaCl). For allantoinase, functionally active enzyme was expressed and purified as described previously (14).…”

Structure of dihydropyrimidinase from Sinorhizobium meliloti CECT4114: New features in an amidohydrolase family member

“…This b-sandwich domain has been identified in other amidohydrolase-related enzymes, such as allantoinase (PDB ID. 3E74; Kim et al, 2009), dihydroorotase (PDB ID. 1XRF; Martin et al, 2005), cytosine deaminase (PDB ID.…”

Cloning, expression, purification, crystallization and preliminary X-ray characterization of allantoinase fromBacillus licheniformisATCC 14580

“…Allantoinase catalyzes the reversible hydrolysis of allantoin into allantoate through the hydrolytic cleavage of the five-membered hydantoin ring (Noguchi et al, 1986;. Allantoin and allantoate, collectively called ureides, play essential roles in nitrogen assimilation, storage and transport (Kim et al, 2009). Biochemical studies using Escherichia coli allantoinase have proved that its activity depends on the presence and the type of metal in the active centre, analogous to what has been observed for other enzymes of this family (Kim et al, 2000;Mulrooney & Hausinger, 2003).…”