Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13
<i>N</i>-Heterocyclic compounds are widely spread in the biosphere, being constituents of alkaloids, cofactors, allelochemicals, and artificial substances. However, the fate of such compounds including a catabolism of hydroxylated pyridines is not yet fully understood. <i>Arthrobac...
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MDPI AG
2020-06-01
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| Series: | Microorganisms |
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| author | Justas Vaitekūnas Renata Gasparavičiūtė Jonita Stankevičiūtė Gintaras Urbelis Rolandas Meškys |
| author_facet | Justas Vaitekūnas Renata Gasparavičiūtė Jonita Stankevičiūtė Gintaras Urbelis Rolandas Meškys |
| author_sort | Justas Vaitekūnas |
| collection | DOAJ |
| description | <i>N</i>-Heterocyclic compounds are widely spread in the biosphere, being constituents of alkaloids, cofactors, allelochemicals, and artificial substances. However, the fate of such compounds including a catabolism of hydroxylated pyridines is not yet fully understood. <i>Arthrobacter</i> sp. IN13 is capable of using 4-hydroxypyridine as a sole source of carbon and energy. Three substrate-inducible proteins were detected by comparing protein expression profiles, and peptide mass fingerprinting was performed using MS/MS. After partial sequencing of the genome, we were able to locate genes encoding 4-hydroxypyridine-inducible proteins and identify the <i>kpi</i> gene cluster consisting of 16 open reading frames. The recombinant expression of genes from this locus in <i>Escherichia coli</i> and <i>Rhodococcus erytropolis</i> SQ1 allowed an elucidation of the biochemical functions of the proteins. We report that in <i>Arthrobacter</i> sp. IN13, the initial hydroxylation of 4-hydroxypyridine is catalyzed by a flavin-dependent monooxygenase (KpiA). A product of the monooxygenase reaction is identified as 3,4-dihydroxypyridine, and a subsequent oxidative opening of the ring is performed by a hypothetical amidohydrolase (KpiC). The 3-(<i>N</i>-formyl)-formiminopyruvate formed in this reaction is further converted by KpiB hydrolase to 3-formylpyruvate. Thus, the degradation of 4-hydroxypyridine in <i>Arthrobacter</i> sp. IN13 was analyzed at genetic and biochemical levels, elucidating this catabolic pathway. |
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| last_indexed | 2024-03-10T19:13:41Z |
| publishDate | 2020-06-01 |
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| series | Microorganisms |
| spelling | doaj.art-12de85d614904c49aecd2e948cb6a0872023-11-20T03:35:06ZengMDPI AGMicroorganisms2076-26072020-06-018688810.3390/microorganisms8060888Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13Justas Vaitekūnas0Renata Gasparavičiūtė1Jonita Stankevičiūtė2Gintaras Urbelis3Rolandas Meškys4Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio al. 7, LT-10257 Vilnius, LithuaniaDepartment of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio al. 7, LT-10257 Vilnius, LithuaniaDepartment of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio al. 7, LT-10257 Vilnius, LithuaniaDepartment of Organic Chemistry, Center for Physical Sciences and Technology, Akademijos 7, LT-08412 Vilnius, LithuaniaDepartment of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Saulėtekio al. 7, LT-10257 Vilnius, Lithuania<i>N</i>-Heterocyclic compounds are widely spread in the biosphere, being constituents of alkaloids, cofactors, allelochemicals, and artificial substances. However, the fate of such compounds including a catabolism of hydroxylated pyridines is not yet fully understood. <i>Arthrobacter</i> sp. IN13 is capable of using 4-hydroxypyridine as a sole source of carbon and energy. Three substrate-inducible proteins were detected by comparing protein expression profiles, and peptide mass fingerprinting was performed using MS/MS. After partial sequencing of the genome, we were able to locate genes encoding 4-hydroxypyridine-inducible proteins and identify the <i>kpi</i> gene cluster consisting of 16 open reading frames. The recombinant expression of genes from this locus in <i>Escherichia coli</i> and <i>Rhodococcus erytropolis</i> SQ1 allowed an elucidation of the biochemical functions of the proteins. We report that in <i>Arthrobacter</i> sp. IN13, the initial hydroxylation of 4-hydroxypyridine is catalyzed by a flavin-dependent monooxygenase (KpiA). A product of the monooxygenase reaction is identified as 3,4-dihydroxypyridine, and a subsequent oxidative opening of the ring is performed by a hypothetical amidohydrolase (KpiC). The 3-(<i>N</i>-formyl)-formiminopyruvate formed in this reaction is further converted by KpiB hydrolase to 3-formylpyruvate. Thus, the degradation of 4-hydroxypyridine in <i>Arthrobacter</i> sp. IN13 was analyzed at genetic and biochemical levels, elucidating this catabolic pathway.https://www.mdpi.com/2076-2607/8/6/8884-hydroxypyridine3,4-dihydroxypyridine4-hydroxypyridine 3-monooxygenase3,4-dihydroxypyridine dioxygenaseextradiol dioxygenaseamidohydrolase |
| spellingShingle | Justas Vaitekūnas Renata Gasparavičiūtė Jonita Stankevičiūtė Gintaras Urbelis Rolandas Meškys Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13 Microorganisms 4-hydroxypyridine 3,4-dihydroxypyridine 4-hydroxypyridine 3-monooxygenase 3,4-dihydroxypyridine dioxygenase extradiol dioxygenase amidohydrolase |
| title | Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13 |
| title_full | Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13 |
| title_fullStr | Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13 |
| title_full_unstemmed | Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13 |
| title_short | Biochemical and Genetic Analysis of 4-Hydroxypyridine Catabolism in <i>Arthrobacter</i> sp. Strain IN13 |
| title_sort | biochemical and genetic analysis of 4 hydroxypyridine catabolism in i arthrobacter i sp strain in13 |
| topic | 4-hydroxypyridine 3,4-dihydroxypyridine 4-hydroxypyridine 3-monooxygenase 3,4-dihydroxypyridine dioxygenase extradiol dioxygenase amidohydrolase |
| url | https://www.mdpi.com/2076-2607/8/6/888 |
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