Solving the Conundrum: Widespread Proteins Annotated for Urea Metabolism in Bacteria Are Carboxyguanidine Deiminases Mediating Nitrogen Assimilation from Guanidine

Nicholas O. Schneider, Lambros J. Tassoulas, Danyun Zeng, Amanda J. Laseke, Nicholas J. Reiter, Lawrence P. Wackett, Martin St Maurice

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Free guanidine is increasingly recognized as a relevant molecule in biological systems. Recently, it was reported that urea carboxylase acts preferentially on guanidine, and consequently, it was considered to participate directly in guanidine biodegradation. Urea carboxylase combines with allophanate hydrolase to comprise the activity of urea amidolyase, an enzyme predominantly found in bacteria and fungi that catalyzes the carboxylation and subsequent hydrolysis of urea to ammonia and carbon dioxide. Here, we demonstrate that urea carboxylase and allophanate hydrolase from Pseudomonas syringae are insufficient to catalyze the decomposition of guanidine. Rather, guanidine is decomposed to ammonia through the combined activities of urea carboxylase, allophanate hydrolase, and two additional proteins of the DUF1989 protein family, expansively annotated as urea carboxylase-associated family proteins. These proteins comprise the subunits of a heterodimeric carboxyguanidine deiminase (CgdAB), which hydrolyzes carboxyguanidine to N-carboxyurea (allophanate). The genes encoding CgdAB colocalize with genes encoding urea carboxylase and allophanate hydrolase. However, 25% of urea carboxylase genes, including all fungal urea amidolyases, do not colocalize with cgdAB. This subset of urea carboxylases correlates with a notable Asp to Asn mutation in the carboxyltransferase active site. Consistent with this observation, we demonstrate that fungal urea amidolyase retains a strong substrate preference for urea. The combined activities of urea carboxylase, carboxyguanidine deiminase and allophanate hydrolase represent a newly recognized pathway for the biodegradation of guanidine. These findings reinforce the relevance of guanidine as a biological metabolite and reveal a broadly distributed group of enzymes that act on guanidine in bacteria.

Original languageEnglish (US)
Pages (from-to)3258-3270
Number of pages13
JournalBiochemistry
Volume59
Issue number35
DOIs
StatePublished - Sep 8 2020

Bibliographical note

Funding Information:
This work was supported by the National Institute of General Medical Sciences of the National Institute of Health under Award Number R15GM097724–01A1 to MStM, USDA National Institute of Food and Agriculture, Agricultural and Food Research Initiative Competitive Program, Ecosystem Services and Agro-Ecosystem Management, grant no. 2019–67019–29403 to LPW and National Institutes of Health Biotechnology training grant (5T32GM008347–27) for LJT. NJR and DZ acknowledge support from the NIH R01GM120572 and the Marquette University Chemistry Department.

Funding Information:
This work was supported by the National Institute of General Medical Sciences of the National Institute of Health under Award Number R15GM097724-01A1 to MStM, USDA National Institute of Food and Agriculture, Agricultural and Food Research Initiative Competitive Program, Ecosystem Services and Agro-Ecosystem Management, grant no. 2019-67019-29403 to LPW and National Institutes of Health Biotechnology training grant (5T32GM008347-27) for LJT. NJR and DZ acknowledge support from the NIH R01GM120572 and the Marquette University Chemistry Department.

Publisher Copyright:
Copyright © 2020 American Chemical Society.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

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