Purine utilization by Klebsiella oxytoca M5al: genes for ring-oxidizing and -opening enzymes.
The enterobacterium Klebsiella oxytoca uses a variety of inorganic and organic nitrogen sources, including purines, nitrogen-rich compounds that are widespread in the biosphere. We have identified a 23-gene cluster that encodes the enzymes for utilizing purines as the sole nitrogen source. Growth and complementation tests with insertion mutants, combined with sequence comparisons, reveal functions for the products of these genes. Here, we report our characterization of 12 genes, one encoding guanine deaminase and the others encoding enzymes for converting (hypo)xanthine to allantoate. Conventionally, xanthine dehydrogenase, a broadly distributed molybdoflavoenzyme, catalyzes sequential hydroxylation reactions to convert hypoxanthine via xanthine to urate. Our results show that these reactions in K. oxytoca are catalyzed by a two-component oxygenase (HpxE-HpxD enzyme) homologous to Rieske nonheme iron aromatic-ring-hydroxylating systems, such as phthalate dioxygenase. Our results also reveal previously undescribed enzymes involved in urate oxidation to allantoin, catalyzed by a flavoprotein monooxygenase (HpxO enzyme), and in allantoin conversion to allantoate, which involves allantoin racemase (HpxA enzyme). The pathway also includes the recently described PuuE allantoinase (HpxB enzyme). The HpxE-HpxD and HpxO enzymes were discovered independently by de la Riva et al. (L. de la Riva, J. Badia, J. Aguilar, R. A. Bender, and L. Baldoma, J. Bacteriol. 190:7892-7903, 2008). Thus, several enzymes in this K. oxytoca purine utilization pathway differ from those in other microorganisms. Isofunctional homologs of these enzymes apparently are encoded by other species, including Acinetobacter, Burkholderia, Pseudomonas, Saccharomyces, and Xanthomonas.
Pope
Scott D
SD
Department of Microbiology, University of California, One Shields Ave., Davis, CA 95616-8665, USA.
Chen
Li-Ling
LL
Stewart
Valley
V
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D006147:Guanine / Q000737:chemistry / Q000378:metabolism; D006148:Guanine Deaminase / Q000235:genetics / Q000378:metabolism; D041121:Klebsiella oxytoca / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011687:Purines / Q000378:metabolism*; D019998:Racemases and Epimerases / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic; D014508:Urea / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D014527:Uric Acid / Q000737:chemistry / Q000378:metabolism; D019820:Xanthine / Q000737:chemistry / Q000378:metabolism; D014968:Xanthine Dehydrogenase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D005656:Fungal Proteins; D011687:Purines; C024081:allantoic acid; D019820:Xanthine; D014527:Uric Acid; D006147:Guanine; D014508:Urea; D049308:Dioxygenases; C503093:xanthine dioxygenase, Aspergillus nidulans; D014968:Xanthine Dehydrogenase; D006148:Guanine Deaminase; D019998:Racemases and Epimerases; C019660:allantoin racemase; C030985:purine