Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production
Abstract Benzylisoquinoline alkaloids (BIAs) are a type of secondary metabolite with clinical application value. (S)-stylopine is a special BIA which contains methylenedioxy bridge structures. CYP719As could catalyze the methylenedioxy bridge-formation on the A or D rings of protoberberine alkaloids...
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| Format: | Article |
| Language: | English |
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BMC
2023-02-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-023-02024-2 |
| _version_ | 1811171538110513152 |
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| author | Xiuyu Liu Xiang Jiao Yatian Cheng Ying Ma Junling Bu Baolong Jin Qishuang Li Zhimin Hu Jinfu Tang Changjiangsheng Lai Jian Wang Guanghong Cui Yun Chen Juan Guo Luqi Huang |
| author_facet | Xiuyu Liu Xiang Jiao Yatian Cheng Ying Ma Junling Bu Baolong Jin Qishuang Li Zhimin Hu Jinfu Tang Changjiangsheng Lai Jian Wang Guanghong Cui Yun Chen Juan Guo Luqi Huang |
| author_sort | Xiuyu Liu |
| collection | DOAJ |
| description | Abstract Benzylisoquinoline alkaloids (BIAs) are a type of secondary metabolite with clinical application value. (S)-stylopine is a special BIA which contains methylenedioxy bridge structures. CYP719As could catalyze the methylenedioxy bridge-formation on the A or D rings of protoberberine alkaloids, while displaying significant substrate regiospecificity. To explore the substrate preference of CYP719As, we cloned and identified five CyCYP719A candidates from Corydalis yanhusuo. Two CyCYP719As (CyCYP719A39 and CyCYP719A42) with high catalytic efficiency for the methylenedioxy bridge-formation on the D or A rings were characterized, respectively. The residues (Leu 294 for CyCYP719A42 and Asp 289 for CyCYP719A39) were identified as the key to controlling the regioselectivity of CYP719As affecting the methylenedioxy bridge-formation on the A or D rings by homology modeling and mutation analysis. Furthermore, for de novo production of BIAs, CyCYP719A39, CyCYP719A42, and their mutants were introduced into the (S)-scoulerine-producing yeast to produce 32 mg/L (S)-stylopine. These results lay a foundation for understanding the structure-function relationship of CYP719A-mediated methylenedioxy bridge-formation and provide yeast strains for the BIAs production by synthetic biology. |
| first_indexed | 2024-04-10T17:15:39Z |
| format | Article |
| id | doaj.art-e73d407b75a3406b8d95dc7029725dd1 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-10T17:15:39Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-e73d407b75a3406b8d95dc7029725dd12023-02-05T12:27:42ZengBMCMicrobial Cell Factories1475-28592023-02-0122111310.1186/s12934-023-02024-2Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo productionXiuyu Liu0Xiang Jiao1Yatian Cheng2Ying Ma3Junling Bu4Baolong Jin5Qishuang Li6Zhimin Hu7Jinfu Tang8Changjiangsheng Lai9Jian Wang10Guanghong Cui11Yun Chen12Juan Guo13Luqi Huang14State Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesDepartment of Biology and Biological Engineering, Chalmers University of TechnologyState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesDepartment of Biology and Biological Engineering, Chalmers University of TechnologyState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesState Key Laboratory of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical SciencesAbstract Benzylisoquinoline alkaloids (BIAs) are a type of secondary metabolite with clinical application value. (S)-stylopine is a special BIA which contains methylenedioxy bridge structures. CYP719As could catalyze the methylenedioxy bridge-formation on the A or D rings of protoberberine alkaloids, while displaying significant substrate regiospecificity. To explore the substrate preference of CYP719As, we cloned and identified five CyCYP719A candidates from Corydalis yanhusuo. Two CyCYP719As (CyCYP719A39 and CyCYP719A42) with high catalytic efficiency for the methylenedioxy bridge-formation on the D or A rings were characterized, respectively. The residues (Leu 294 for CyCYP719A42 and Asp 289 for CyCYP719A39) were identified as the key to controlling the regioselectivity of CYP719As affecting the methylenedioxy bridge-formation on the A or D rings by homology modeling and mutation analysis. Furthermore, for de novo production of BIAs, CyCYP719A39, CyCYP719A42, and their mutants were introduced into the (S)-scoulerine-producing yeast to produce 32 mg/L (S)-stylopine. These results lay a foundation for understanding the structure-function relationship of CYP719A-mediated methylenedioxy bridge-formation and provide yeast strains for the BIAs production by synthetic biology.https://doi.org/10.1186/s12934-023-02024-2CyCYP719AsBenzylisoquinoline alkaloids (BIAs)Corydalis yanhusuoMethylenedioxy bridge-formationRegiospecificitySynthetic biology |
| spellingShingle | Xiuyu Liu Xiang Jiao Yatian Cheng Ying Ma Junling Bu Baolong Jin Qishuang Li Zhimin Hu Jinfu Tang Changjiangsheng Lai Jian Wang Guanghong Cui Yun Chen Juan Guo Luqi Huang Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production Microbial Cell Factories CyCYP719As Benzylisoquinoline alkaloids (BIAs) Corydalis yanhusuo Methylenedioxy bridge-formation Regiospecificity Synthetic biology |
| title | Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production |
| title_full | Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production |
| title_fullStr | Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production |
| title_full_unstemmed | Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production |
| title_short | Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production |
| title_sort | structure function analysis of cyp719as involved in methylenedioxy bridge formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production |
| topic | CyCYP719As Benzylisoquinoline alkaloids (BIAs) Corydalis yanhusuo Methylenedioxy bridge-formation Regiospecificity Synthetic biology |
| url | https://doi.org/10.1186/s12934-023-02024-2 |
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