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A novel formamidase is required for riboflavin biosynthesis in invasive bacteria
Biosynthesis of riboflavin (RF), the precursor of the redox cofactors FMN and FAD, was thought to be well understood in bacteria, with all the pathway enzymes presumed to be known and essential. Our previous research has challenged this view by showing that, in the bacterium Sinorhizobium meliloti,...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9478397/ https://www.ncbi.nlm.nih.gov/pubmed/35970388 http://dx.doi.org/10.1016/j.jbc.2022.102377 |
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author | Yurgel, Svetlana N. Johnson, Skylar A. Rice, Jennifer Sa, Na Bailes, Clayton Baumgartner, John Pitzer, Josh E. Roop, R. Martin Roje, Sanja |
author_facet | Yurgel, Svetlana N. Johnson, Skylar A. Rice, Jennifer Sa, Na Bailes, Clayton Baumgartner, John Pitzer, Josh E. Roop, R. Martin Roje, Sanja |
author_sort | Yurgel, Svetlana N. |
collection | PubMed |
description | Biosynthesis of riboflavin (RF), the precursor of the redox cofactors FMN and FAD, was thought to be well understood in bacteria, with all the pathway enzymes presumed to be known and essential. Our previous research has challenged this view by showing that, in the bacterium Sinorhizobium meliloti, deletion of the ribBA gene encoding the enzyme that catalyzes the initial steps on the RF biosynthesis pathway only causes a reduction in flavin secretion rather than RF auxotrophy. This finding led us to hypothesize that RibBA participates in the biosynthesis of flavins destined for secretion, whereas S. meliloti has another enzyme that performs this function for internal cellular metabolism. Here, we identify and biochemically characterize a novel formamidase (SMc02977) involved in the production of RF for intracellular functions in S. meliloti. This catalyst, which we named Sm-BrbF, releases formate from the early RF precursor 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate to yield 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate. We show that homologs of this enzyme are present in many bacteria, are highly abundant in the Rhizobiales order, and that sequence homologs from Brucella abortus and Liberobacter solanacearum complement the RF auxotrophy of the Sm1021ΔSMc02977 mutant. Furthermore, we show that the B. abortus enzyme (Bab2_0247, Ba-BrbF) is also an 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate formamidase, and that the bab2_0247 mutant is a RF auxotroph exhibiting a lower level of intracellular infection than the wildtype strain. Finally, we show that Sm-BrbF and Ba-BrbF directly interact with other RF biosynthesis pathway enzymes. Together, our results provide novel insight into the intricacies of RF biosynthesis in bacteria. |
format | Online Article Text |
id | pubmed-9478397 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-94783972022-09-22 A novel formamidase is required for riboflavin biosynthesis in invasive bacteria Yurgel, Svetlana N. Johnson, Skylar A. Rice, Jennifer Sa, Na Bailes, Clayton Baumgartner, John Pitzer, Josh E. Roop, R. Martin Roje, Sanja J Biol Chem Research Article Biosynthesis of riboflavin (RF), the precursor of the redox cofactors FMN and FAD, was thought to be well understood in bacteria, with all the pathway enzymes presumed to be known and essential. Our previous research has challenged this view by showing that, in the bacterium Sinorhizobium meliloti, deletion of the ribBA gene encoding the enzyme that catalyzes the initial steps on the RF biosynthesis pathway only causes a reduction in flavin secretion rather than RF auxotrophy. This finding led us to hypothesize that RibBA participates in the biosynthesis of flavins destined for secretion, whereas S. meliloti has another enzyme that performs this function for internal cellular metabolism. Here, we identify and biochemically characterize a novel formamidase (SMc02977) involved in the production of RF for intracellular functions in S. meliloti. This catalyst, which we named Sm-BrbF, releases formate from the early RF precursor 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate to yield 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate. We show that homologs of this enzyme are present in many bacteria, are highly abundant in the Rhizobiales order, and that sequence homologs from Brucella abortus and Liberobacter solanacearum complement the RF auxotrophy of the Sm1021ΔSMc02977 mutant. Furthermore, we show that the B. abortus enzyme (Bab2_0247, Ba-BrbF) is also an 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5′-phosphate formamidase, and that the bab2_0247 mutant is a RF auxotroph exhibiting a lower level of intracellular infection than the wildtype strain. Finally, we show that Sm-BrbF and Ba-BrbF directly interact with other RF biosynthesis pathway enzymes. Together, our results provide novel insight into the intricacies of RF biosynthesis in bacteria. American Society for Biochemistry and Molecular Biology 2022-08-13 /pmc/articles/PMC9478397/ /pubmed/35970388 http://dx.doi.org/10.1016/j.jbc.2022.102377 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Yurgel, Svetlana N. Johnson, Skylar A. Rice, Jennifer Sa, Na Bailes, Clayton Baumgartner, John Pitzer, Josh E. Roop, R. Martin Roje, Sanja A novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
title | A novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
title_full | A novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
title_fullStr | A novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
title_full_unstemmed | A novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
title_short | A novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
title_sort | novel formamidase is required for riboflavin biosynthesis in invasive bacteria |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9478397/ https://www.ncbi.nlm.nih.gov/pubmed/35970388 http://dx.doi.org/10.1016/j.jbc.2022.102377 |
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