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Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold
Plant-originated natural products are important drug sources. However, total biosynthesis of these compounds is often not achievable due to their uncharacterized, lengthy biosynthetic pathways. In nature, phenethylisoquinoline alkaloids (PIAs) such as colchicine are biosynthesized via a common precu...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
KeAi Publishing
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10534260/ https://www.ncbi.nlm.nih.gov/pubmed/37781172 http://dx.doi.org/10.1016/j.synbio.2023.09.003 |
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author | Zhang, Yuhao Liu, Wan-Qiu Li, Jian |
author_facet | Zhang, Yuhao Liu, Wan-Qiu Li, Jian |
author_sort | Zhang, Yuhao |
collection | PubMed |
description | Plant-originated natural products are important drug sources. However, total biosynthesis of these compounds is often not achievable due to their uncharacterized, lengthy biosynthetic pathways. In nature, phenethylisoquinoline alkaloids (PIAs) such as colchicine are biosynthesized via a common precursor 6,7-dihydroxy-1-(4-hydroxyphenylethyl)-1,2,3,4-tetrahydroisoquinoline (i.e., phenethylisoquinoline scaffold, PIAS). PIAS is naturally synthesized in plants by using two upstream substrates (l-phenylalanine and l-tyrosine) catalyzed by eight enzymes. To shorten this native pathway, here we designed an artificial route to synthesize PIAS with two enzymatic steps from two alternative substrates of 3-(4-hydroxyphenyl) propanol (4-HPP) and dopamine. In the two-step bioconversion, an alcohol dehydrogenase selected from yeast (i.e., ADH7) was able to oxidize its non-native alcohol substrate 4-HPP to form the corresponding aldehyde product, which was then condensed with dopamine by the (S)-norcoclaurine synthase (NCS) to synthesize PIAS. After optimization, the final enzymatic reaction system was successfully scaled up by 200 times from 50 μL to 10 mL, generating 5.4 mM of PIAS. We envision that this study will provide an easy and sustainable approach to produce PIAS and thus lay the foundation for large-scale production of PIAS-derived natural products. |
format | Online Article Text |
id | pubmed-10534260 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | KeAi Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-105342602023-09-29 Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold Zhang, Yuhao Liu, Wan-Qiu Li, Jian Synth Syst Biotechnol Original Research Article Plant-originated natural products are important drug sources. However, total biosynthesis of these compounds is often not achievable due to their uncharacterized, lengthy biosynthetic pathways. In nature, phenethylisoquinoline alkaloids (PIAs) such as colchicine are biosynthesized via a common precursor 6,7-dihydroxy-1-(4-hydroxyphenylethyl)-1,2,3,4-tetrahydroisoquinoline (i.e., phenethylisoquinoline scaffold, PIAS). PIAS is naturally synthesized in plants by using two upstream substrates (l-phenylalanine and l-tyrosine) catalyzed by eight enzymes. To shorten this native pathway, here we designed an artificial route to synthesize PIAS with two enzymatic steps from two alternative substrates of 3-(4-hydroxyphenyl) propanol (4-HPP) and dopamine. In the two-step bioconversion, an alcohol dehydrogenase selected from yeast (i.e., ADH7) was able to oxidize its non-native alcohol substrate 4-HPP to form the corresponding aldehyde product, which was then condensed with dopamine by the (S)-norcoclaurine synthase (NCS) to synthesize PIAS. After optimization, the final enzymatic reaction system was successfully scaled up by 200 times from 50 μL to 10 mL, generating 5.4 mM of PIAS. We envision that this study will provide an easy and sustainable approach to produce PIAS and thus lay the foundation for large-scale production of PIAS-derived natural products. KeAi Publishing 2023-09-22 /pmc/articles/PMC10534260/ /pubmed/37781172 http://dx.doi.org/10.1016/j.synbio.2023.09.003 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Research Article Zhang, Yuhao Liu, Wan-Qiu Li, Jian Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
title | Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
title_full | Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
title_fullStr | Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
title_full_unstemmed | Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
title_short | Constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
title_sort | constructing an artificial short route for cell-free biosynthesis of the phenethylisoquinoline scaffold |
topic | Original Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10534260/ https://www.ncbi.nlm.nih.gov/pubmed/37781172 http://dx.doi.org/10.1016/j.synbio.2023.09.003 |
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