Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1
Abstract Background Folic acid (FA) is a synthetic vitamin (B9) and the oxidized form of a metabolic cofactor that is essential for life. Although the biosynthetic mechanisms of FA are established, its environmental degradation mechanism has not been fully elucidated. The present study aimed to iden...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-09-01
|
| Series: | BMC Microbiology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12866-022-02643-6 |
| _version_ | 1818024830247632896 |
|---|---|
| author | Yungmi You Yuki Doi Norifumi Maeda Shunsuke Masuo Norio Takeshita Naoki Takaya |
| author_facet | Yungmi You Yuki Doi Norifumi Maeda Shunsuke Masuo Norio Takeshita Naoki Takaya |
| author_sort | Yungmi You |
| collection | DOAJ |
| description | Abstract Background Folic acid (FA) is a synthetic vitamin (B9) and the oxidized form of a metabolic cofactor that is essential for life. Although the biosynthetic mechanisms of FA are established, its environmental degradation mechanism has not been fully elucidated. The present study aimed to identify bacteria in soil that degrade FA and the mechanisms involved. Results We isolated the soil bacterium Variovorax sp. F1 from sampled weed rhizospheres in a grassland and investigated its FA degradation mechanism. Cultured Variovorax sp. F1 rapidly degraded FA to pteroic acid (PA), indicating that FA hydrolysis to PA and glutamate. We cloned the carboxypeptidase G (CPG) gene and found widely distributed paralogs within the Variovorax genus. Recombinant CPG preferred FA and deaminofolic acid as substrates, indicating its involvement in FA degradation by Variovorax. Prolonged culture of Variovorax sp. F1 resulted in decreased rates of deaminofolic acid (DFA) and deaminopteroic acid (DPA) accumulation. This indicated that the deamination reaction also comprised a route of FA degradation. We also identified an F1 gene that was orthologous to the pterin deaminase gene (Arad3529) of Agrobacterium radiobacter. The encoded protein deaminated FA and PA to DFA and DPA, which was consistent with the deamination activity of FA and PA in bacterial cell-free extracts. Conclusion We discovered that the two enzymes required for FA degradation pathways in isolates of Variovorax sp. F1 comprise CPG and pterin deaminase, and that DFA and PA are intermediates in the generation of DPA. |
| first_indexed | 2024-12-10T04:06:27Z |
| format | Article |
| id | doaj.art-ddbec97c52d64a98a3ed6a1e6e8da9c8 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-12-10T04:06:27Z |
| publishDate | 2022-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj.art-ddbec97c52d64a98a3ed6a1e6e8da9c82022-12-22T02:02:50ZengBMCBMC Microbiology1471-21802022-09-0122111310.1186/s12866-022-02643-6Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1Yungmi You0Yuki Doi1Norifumi Maeda2Shunsuke Masuo3Norio Takeshita4Naoki Takaya5Faculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of TsukubaFaculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of TsukubaFaculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of TsukubaFaculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of TsukubaFaculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of TsukubaFaculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of TsukubaAbstract Background Folic acid (FA) is a synthetic vitamin (B9) and the oxidized form of a metabolic cofactor that is essential for life. Although the biosynthetic mechanisms of FA are established, its environmental degradation mechanism has not been fully elucidated. The present study aimed to identify bacteria in soil that degrade FA and the mechanisms involved. Results We isolated the soil bacterium Variovorax sp. F1 from sampled weed rhizospheres in a grassland and investigated its FA degradation mechanism. Cultured Variovorax sp. F1 rapidly degraded FA to pteroic acid (PA), indicating that FA hydrolysis to PA and glutamate. We cloned the carboxypeptidase G (CPG) gene and found widely distributed paralogs within the Variovorax genus. Recombinant CPG preferred FA and deaminofolic acid as substrates, indicating its involvement in FA degradation by Variovorax. Prolonged culture of Variovorax sp. F1 resulted in decreased rates of deaminofolic acid (DFA) and deaminopteroic acid (DPA) accumulation. This indicated that the deamination reaction also comprised a route of FA degradation. We also identified an F1 gene that was orthologous to the pterin deaminase gene (Arad3529) of Agrobacterium radiobacter. The encoded protein deaminated FA and PA to DFA and DPA, which was consistent with the deamination activity of FA and PA in bacterial cell-free extracts. Conclusion We discovered that the two enzymes required for FA degradation pathways in isolates of Variovorax sp. F1 comprise CPG and pterin deaminase, and that DFA and PA are intermediates in the generation of DPA.https://doi.org/10.1186/s12866-022-02643-6Vitamin B9Pterin deaminasePteroic acidDeaminofolic acidVariovorax |
| spellingShingle | Yungmi You Yuki Doi Norifumi Maeda Shunsuke Masuo Norio Takeshita Naoki Takaya Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1 BMC Microbiology Vitamin B9 Pterin deaminase Pteroic acid Deaminofolic acid Variovorax |
| title | Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1 |
| title_full | Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1 |
| title_fullStr | Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1 |
| title_full_unstemmed | Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1 |
| title_short | Carboxypeptidase G and pterin deaminase metabolic pathways degrade folic acid in Variovorax sp. F1 |
| title_sort | carboxypeptidase g and pterin deaminase metabolic pathways degrade folic acid in variovorax sp f1 |
| topic | Vitamin B9 Pterin deaminase Pteroic acid Deaminofolic acid Variovorax |
| url | https://doi.org/10.1186/s12866-022-02643-6 |
| work_keys_str_mv | AT yungmiyou carboxypeptidasegandpterindeaminasemetabolicpathwaysdegradefolicacidinvariovoraxspf1 AT yukidoi carboxypeptidasegandpterindeaminasemetabolicpathwaysdegradefolicacidinvariovoraxspf1 AT norifumimaeda carboxypeptidasegandpterindeaminasemetabolicpathwaysdegradefolicacidinvariovoraxspf1 AT shunsukemasuo carboxypeptidasegandpterindeaminasemetabolicpathwaysdegradefolicacidinvariovoraxspf1 AT noriotakeshita carboxypeptidasegandpterindeaminasemetabolicpathwaysdegradefolicacidinvariovoraxspf1 AT naokitakaya carboxypeptidasegandpterindeaminasemetabolicpathwaysdegradefolicacidinvariovoraxspf1 |