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Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast
The development of medical applications exploiting the broad bioactivities of the diterpene therapeutic triptolide from Tripterygium wilfordii is limited by low extraction yields from the native plant. Furthermore, the extraordinarily high structural complexity prevents an economically attractive en...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152743/ https://www.ncbi.nlm.nih.gov/pubmed/28608823 http://dx.doi.org/10.3390/molecules22060981 |
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author | Forman, Victor Callari, Roberta Folly, Christophe Heider, Harald Hamberger, Björn |
author_facet | Forman, Victor Callari, Roberta Folly, Christophe Heider, Harald Hamberger, Björn |
author_sort | Forman, Victor |
collection | PubMed |
description | The development of medical applications exploiting the broad bioactivities of the diterpene therapeutic triptolide from Tripterygium wilfordii is limited by low extraction yields from the native plant. Furthermore, the extraordinarily high structural complexity prevents an economically attractive enantioselective total synthesis. An alternative production route of triptolide through engineered Saccharomyces cerevisiae (yeast) could provide a sustainable source of triptolide. A potential intermediate in the unknown biosynthetic route to triptolide is the diterpene dehydroabietic acid. Here, we report a biosynthetic route to dehydroabietic acid by transient expression of enzymes from T. wilfordii and Sitka spruce (Picea sitchensis) in Nicotiana benthamiana. The combination of diterpene synthases TwTPS9, TwTPS27, and cytochromes P450 PsCYP720B4 yielded dehydroabietic acid and a novel analog, tentatively identified as ‘miltiradienic acid’. This biosynthetic pathway was reassembled in a yeast strain engineered for increased yields of the pathway intermediates, the diterpene olefins miltiradiene and dehydroabietadiene. Introduction in that strain of PsCYP720B4 in combination with two alternative NADPH-dependent cytochrome P450 reductases resulted in scalable in vivo production of dehydroabietic acid and its analog from glucose. Approaching future elucidation of the remaining biosynthetic steps to triptolide, our findings may provide an independent platform for testing of additional recombinant candidate genes, and ultimately pave the way to biotechnological production of the high value diterpenoid therapeutic. |
format | Online Article Text |
id | pubmed-6152743 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-61527432018-11-13 Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast Forman, Victor Callari, Roberta Folly, Christophe Heider, Harald Hamberger, Björn Molecules Article The development of medical applications exploiting the broad bioactivities of the diterpene therapeutic triptolide from Tripterygium wilfordii is limited by low extraction yields from the native plant. Furthermore, the extraordinarily high structural complexity prevents an economically attractive enantioselective total synthesis. An alternative production route of triptolide through engineered Saccharomyces cerevisiae (yeast) could provide a sustainable source of triptolide. A potential intermediate in the unknown biosynthetic route to triptolide is the diterpene dehydroabietic acid. Here, we report a biosynthetic route to dehydroabietic acid by transient expression of enzymes from T. wilfordii and Sitka spruce (Picea sitchensis) in Nicotiana benthamiana. The combination of diterpene synthases TwTPS9, TwTPS27, and cytochromes P450 PsCYP720B4 yielded dehydroabietic acid and a novel analog, tentatively identified as ‘miltiradienic acid’. This biosynthetic pathway was reassembled in a yeast strain engineered for increased yields of the pathway intermediates, the diterpene olefins miltiradiene and dehydroabietadiene. Introduction in that strain of PsCYP720B4 in combination with two alternative NADPH-dependent cytochrome P450 reductases resulted in scalable in vivo production of dehydroabietic acid and its analog from glucose. Approaching future elucidation of the remaining biosynthetic steps to triptolide, our findings may provide an independent platform for testing of additional recombinant candidate genes, and ultimately pave the way to biotechnological production of the high value diterpenoid therapeutic. MDPI 2017-06-13 /pmc/articles/PMC6152743/ /pubmed/28608823 http://dx.doi.org/10.3390/molecules22060981 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Forman, Victor Callari, Roberta Folly, Christophe Heider, Harald Hamberger, Björn Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast |
title | Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast |
title_full | Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast |
title_fullStr | Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast |
title_full_unstemmed | Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast |
title_short | Production of Putative Diterpene Carboxylic Acid Intermediates of Triptolide in Yeast |
title_sort | production of putative diterpene carboxylic acid intermediates of triptolide in yeast |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152743/ https://www.ncbi.nlm.nih.gov/pubmed/28608823 http://dx.doi.org/10.3390/molecules22060981 |
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