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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...

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Detalles Bibliográficos
Autores principales: Zhang, Yuhao, Liu, Wan-Qiu, Li, Jian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: KeAi Publishing 2023
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.
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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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