Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis
Abstract Background Amaryllidaceae alkaloids (AAs) are a large group of plant-specialized metabolites displaying an array of biological and pharmacological properties. Previous investigations on AA biosynthesis have revealed that all AAs share a common precursor, norbelladine, presumably synthesized...
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BMC
2018-12-01
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| Series: | BMC Plant Biology |
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| Online Access: | http://link.springer.com/article/10.1186/s12870-018-1570-4 |
| _version_ | 1818061348300390400 |
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| author | Aparna Singh Marie-Ange Massicotte Ariane Garand Laurence Tousignant Vincent Ouellette Gervais Bérubé Isabel Desgagné-Penix |
| author_facet | Aparna Singh Marie-Ange Massicotte Ariane Garand Laurence Tousignant Vincent Ouellette Gervais Bérubé Isabel Desgagné-Penix |
| author_sort | Aparna Singh |
| collection | DOAJ |
| description | Abstract Background Amaryllidaceae alkaloids (AAs) are a large group of plant-specialized metabolites displaying an array of biological and pharmacological properties. Previous investigations on AA biosynthesis have revealed that all AAs share a common precursor, norbelladine, presumably synthesized by an enzyme catalyzing a Mannich reaction involving the condensation of tyramine and 3,4-dihydroxybenzaldehyde. Similar reactions have been reported. Specifically, norcoclaurine synthase (NCS) which catalyzes the condensation of dopamine and 4-hydroxyphenylacetaldehyde as the first step in benzylisoquinoline alkaloid biosynthesis. Results With the availability of wild daffodil (Narcissus pseudonarcissus) database, a transcriptome-mining search was performed for NCS orthologs. A candidate gene sequence was identified and named norbelladine synthase (NBS). NpNBS encodes for a small protein of 19 kDa with an anticipated pI of 5.5. Phylogenetic analysis showed that NpNBS belongs to a unique clade of PR10/Bet v1 proteins and shared 41% amino acid identity to opium poppy NCS1. Expression of NpNBS cDNA in Escherichia coli produced a recombinant enzyme able to condense tyramine and 3,4-DHBA into norbelladine as determined by high-resolution tandem mass spectrometry. Conclusions Here, we describe a novel enzyme catalyzing the first committed step of AA biosynthesis, which will facilitate the establishment of metabolic engineering and synthetic biology platforms for the production of AAs. |
| first_indexed | 2024-12-10T13:46:53Z |
| format | Article |
| id | doaj.art-221b37f22bf6433b85fb8a9e7285416d |
| institution | Directory Open Access Journal |
| issn | 1471-2229 |
| language | English |
| last_indexed | 2024-12-10T13:46:53Z |
| publishDate | 2018-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Plant Biology |
| spelling | doaj.art-221b37f22bf6433b85fb8a9e7285416d2022-12-22T01:46:21ZengBMCBMC Plant Biology1471-22292018-12-0118111210.1186/s12870-018-1570-4Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesisAparna Singh0Marie-Ange Massicotte1Ariane Garand2Laurence Tousignant3Vincent Ouellette4Gervais Bérubé5Isabel Desgagné-Penix6Department of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresDepartment of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresDepartment of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresDepartment of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresDepartment of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresDepartment of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresDepartment of Chemistry, Biochemistry and Physics, Université du Québec à Trois-RivièresAbstract Background Amaryllidaceae alkaloids (AAs) are a large group of plant-specialized metabolites displaying an array of biological and pharmacological properties. Previous investigations on AA biosynthesis have revealed that all AAs share a common precursor, norbelladine, presumably synthesized by an enzyme catalyzing a Mannich reaction involving the condensation of tyramine and 3,4-dihydroxybenzaldehyde. Similar reactions have been reported. Specifically, norcoclaurine synthase (NCS) which catalyzes the condensation of dopamine and 4-hydroxyphenylacetaldehyde as the first step in benzylisoquinoline alkaloid biosynthesis. Results With the availability of wild daffodil (Narcissus pseudonarcissus) database, a transcriptome-mining search was performed for NCS orthologs. A candidate gene sequence was identified and named norbelladine synthase (NBS). NpNBS encodes for a small protein of 19 kDa with an anticipated pI of 5.5. Phylogenetic analysis showed that NpNBS belongs to a unique clade of PR10/Bet v1 proteins and shared 41% amino acid identity to opium poppy NCS1. Expression of NpNBS cDNA in Escherichia coli produced a recombinant enzyme able to condense tyramine and 3,4-DHBA into norbelladine as determined by high-resolution tandem mass spectrometry. Conclusions Here, we describe a novel enzyme catalyzing the first committed step of AA biosynthesis, which will facilitate the establishment of metabolic engineering and synthetic biology platforms for the production of AAs.http://link.springer.com/article/10.1186/s12870-018-1570-4Amaryllidaceae alkaloidPathogenesis related protein 10Alkaloid biosynthesisNarcissus pseudonarcissusNorcoclaurine synthaseNorbelladine |
| spellingShingle | Aparna Singh Marie-Ange Massicotte Ariane Garand Laurence Tousignant Vincent Ouellette Gervais Bérubé Isabel Desgagné-Penix Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis BMC Plant Biology Amaryllidaceae alkaloid Pathogenesis related protein 10 Alkaloid biosynthesis Narcissus pseudonarcissus Norcoclaurine synthase Norbelladine |
| title | Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis |
| title_full | Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis |
| title_fullStr | Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis |
| title_full_unstemmed | Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis |
| title_short | Cloning and characterization of norbelladine synthase catalyzing the first committed reaction in Amaryllidaceae alkaloid biosynthesis |
| title_sort | cloning and characterization of norbelladine synthase catalyzing the first committed reaction in amaryllidaceae alkaloid biosynthesis |
| topic | Amaryllidaceae alkaloid Pathogenesis related protein 10 Alkaloid biosynthesis Narcissus pseudonarcissus Norcoclaurine synthase Norbelladine |
| url | http://link.springer.com/article/10.1186/s12870-018-1570-4 |
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