Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity
The 6-phosphogluconate dehydrogenase superfamily oxidize and reduce a wide range of substrates, making their functional annotation challenging. Ketolacid reductoisomerase (KARI), encoded by the ilvC gene in branched-chain amino acids biosynthesis, is a promiscuous reductase enzyme within this supe...
| Autores principales: | , , , |
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| Formato: | Artículo |
| Lenguaje: | inglés |
| Publicado: |
2014
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| Acceso en línea: | http://eprints.uanl.mx/15043/1/204.pdf |
| _version_ | 1824414235744010240 |
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| author | Verdel Aranda, Karina López Cortina, Susana Hodgson, David A. Barona Gómez, Francisco |
| author_facet | Verdel Aranda, Karina López Cortina, Susana Hodgson, David A. Barona Gómez, Francisco |
| author_sort | Verdel Aranda, Karina |
| collection | Repositorio Institucional |
| description | The 6-phosphogluconate dehydrogenase superfamily oxidize and reduce a wide range of substrates, making their functional annotation challenging. Ketolacid reductoisomerase (KARI), encoded by the ilvC
gene in branched-chain amino acids biosynthesis, is
a promiscuous reductase enzyme within this superfamily.
Here, we obtain steady-state enzyme kinetic parameters for 10 IlvC homologues from the genera Streptomyces and Corynebacterium, upon eight selected chemically diverse substrates, including some not normally recognized by enzymes of this superfamily. This biochemical data suggested a Streptomyces biosynthetic interlock between proline and the branched-chain amino acids, mediated by
enzyme substrate promiscuity, which was confirmed via mutagenesis and complementation analyses of the proC, ilvC1 and ilvC2 genes in Streptomyces coelicolor. Moreover, both ilvC orthologues and paralogues were analysed, such that the relationship between gene duplication and functional diversification
could be explored. The KARI paralogues present in S. coelicolor and Streptomyces lividans, despite their conserved high sequence identity (97%), were shown to be more promiscuous, suggesting a recent functional diversification. In contrast, the KARI paralogue from Streptomyces viridifaciens showed selectivity towards the synthesis of valine precursors, explaining its recruitment within the biosynthetic gene cluster of valanimycin. These results allowed us to assess substrate promiscuity indices as a tool to annotate new molecular functions with metabolic implications. |
| format | Article |
| id | eprints-15043 |
| institution | UANL |
| language | English |
| publishDate | 2014 |
| record_format | eprints |
| spelling | eprints-150432019-08-21T18:47:23Z http://eprints.uanl.mx/15043/ Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity Verdel Aranda, Karina López Cortina, Susana Hodgson, David A. Barona Gómez, Francisco The 6-phosphogluconate dehydrogenase superfamily oxidize and reduce a wide range of substrates, making their functional annotation challenging. Ketolacid reductoisomerase (KARI), encoded by the ilvC gene in branched-chain amino acids biosynthesis, is a promiscuous reductase enzyme within this superfamily. Here, we obtain steady-state enzyme kinetic parameters for 10 IlvC homologues from the genera Streptomyces and Corynebacterium, upon eight selected chemically diverse substrates, including some not normally recognized by enzymes of this superfamily. This biochemical data suggested a Streptomyces biosynthetic interlock between proline and the branched-chain amino acids, mediated by enzyme substrate promiscuity, which was confirmed via mutagenesis and complementation analyses of the proC, ilvC1 and ilvC2 genes in Streptomyces coelicolor. Moreover, both ilvC orthologues and paralogues were analysed, such that the relationship between gene duplication and functional diversification could be explored. The KARI paralogues present in S. coelicolor and Streptomyces lividans, despite their conserved high sequence identity (97%), were shown to be more promiscuous, suggesting a recent functional diversification. In contrast, the KARI paralogue from Streptomyces viridifaciens showed selectivity towards the synthesis of valine precursors, explaining its recruitment within the biosynthetic gene cluster of valanimycin. These results allowed us to assess substrate promiscuity indices as a tool to annotate new molecular functions with metabolic implications. 2014 Article PeerReviewed text en cc_by_nc_nd http://eprints.uanl.mx/15043/1/204.pdf http://eprints.uanl.mx/15043/1.haspreviewThumbnailVersion/204.pdf Verdel Aranda, Karina y López Cortina, Susana y Hodgson, David A. y Barona Gómez, Francisco (2014) Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity. Microbial Biotechnology, 8 (2). pp. 239-252. ISSN 17517915 http://doi.org/10.1111/1751-7915.12175 doi:10.1111/1751-7915.12175 |
| spellingShingle | Verdel Aranda, Karina López Cortina, Susana Hodgson, David A. Barona Gómez, Francisco Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| thumbnail | https://rediab.uanl.mx/themes/sandal5/images/online.png |
| title | Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| title_full | Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| title_fullStr | Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| title_full_unstemmed | Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| title_short | Molecular annotation of ketol-acid reductoisomerases fromStreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| title_sort | molecular annotation of ketol acid reductoisomerases fromstreptomycesreveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity |
| url | http://eprints.uanl.mx/15043/1/204.pdf |
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