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Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae
Triterpenoids are plant specialized metabolites with various pharmacological activities. They are widely distributed in higher plants, such as legumes. Because of their low accumulation in plants, there is a need for improving triterpenoid production. Cytochrome P450 monooxygenases (CYPs) play criti...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8716914/ https://www.ncbi.nlm.nih.gov/pubmed/34975947 http://dx.doi.org/10.3389/fpls.2021.762546 |
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author | Istiandari, Pramesti Yasumoto, Shuhei Srisawat, Pisanee Tamura, Keita Chikugo, Ayaka Suzuki, Hideyuki Seki, Hikaru Fukushima, Ery Odette Muranaka, Toshiya |
author_facet | Istiandari, Pramesti Yasumoto, Shuhei Srisawat, Pisanee Tamura, Keita Chikugo, Ayaka Suzuki, Hideyuki Seki, Hikaru Fukushima, Ery Odette Muranaka, Toshiya |
author_sort | Istiandari, Pramesti |
collection | PubMed |
description | Triterpenoids are plant specialized metabolites with various pharmacological activities. They are widely distributed in higher plants, such as legumes. Because of their low accumulation in plants, there is a need for improving triterpenoid production. Cytochrome P450 monooxygenases (CYPs) play critical roles in the structural diversification of triterpenoids. To perform site-specific oxidations, CYPs require the electrons that are transferred by NADPH-cytochrome P450 reductase (CPR). Plants possess two main CPR classes, class I and class II. CPR classes I and II have been reported to be responsible for primary and specialized (secondary) metabolism, respectively. In this study, we first analyzed the CPR expression level of three legumes species, Medicago truncatula, Lotus japonicus, and Glycyrrhiza uralensis, showing that the expression level of CPR class I was lower and more stable, while that of CPR class II was higher in almost all the samples. We then co-expressed different combinations of CYP716As and CYP72As with different CPR classes from these three legumes in transgenic yeast. We found that CYP716As worked better with CPR-I from the same species, while CYP72As worked better with any CPR-IIs. Using engineered yeast strains, CYP88D6 paired with class II GuCPR produced the highest level of 11-oxo-β-amyrin, the important precursor of high-value metabolites glycyrrhizin. This study provides insight into co-expressing genes from legumes for heterologous production of triterpenoids in yeast. |
format | Online Article Text |
id | pubmed-8716914 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-87169142021-12-31 Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae Istiandari, Pramesti Yasumoto, Shuhei Srisawat, Pisanee Tamura, Keita Chikugo, Ayaka Suzuki, Hideyuki Seki, Hikaru Fukushima, Ery Odette Muranaka, Toshiya Front Plant Sci Plant Science Triterpenoids are plant specialized metabolites with various pharmacological activities. They are widely distributed in higher plants, such as legumes. Because of their low accumulation in plants, there is a need for improving triterpenoid production. Cytochrome P450 monooxygenases (CYPs) play critical roles in the structural diversification of triterpenoids. To perform site-specific oxidations, CYPs require the electrons that are transferred by NADPH-cytochrome P450 reductase (CPR). Plants possess two main CPR classes, class I and class II. CPR classes I and II have been reported to be responsible for primary and specialized (secondary) metabolism, respectively. In this study, we first analyzed the CPR expression level of three legumes species, Medicago truncatula, Lotus japonicus, and Glycyrrhiza uralensis, showing that the expression level of CPR class I was lower and more stable, while that of CPR class II was higher in almost all the samples. We then co-expressed different combinations of CYP716As and CYP72As with different CPR classes from these three legumes in transgenic yeast. We found that CYP716As worked better with CPR-I from the same species, while CYP72As worked better with any CPR-IIs. Using engineered yeast strains, CYP88D6 paired with class II GuCPR produced the highest level of 11-oxo-β-amyrin, the important precursor of high-value metabolites glycyrrhizin. This study provides insight into co-expressing genes from legumes for heterologous production of triterpenoids in yeast. Frontiers Media S.A. 2021-12-16 /pmc/articles/PMC8716914/ /pubmed/34975947 http://dx.doi.org/10.3389/fpls.2021.762546 Text en Copyright © 2021 Istiandari, Yasumoto, Srisawat, Tamura, Chikugo, Suzuki, Seki, Fukushima and Muranaka. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Istiandari, Pramesti Yasumoto, Shuhei Srisawat, Pisanee Tamura, Keita Chikugo, Ayaka Suzuki, Hideyuki Seki, Hikaru Fukushima, Ery Odette Muranaka, Toshiya Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae |
title | Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae |
title_full | Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae |
title_fullStr | Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae |
title_full_unstemmed | Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae |
title_short | Comparative Analysis of NADPH-Cytochrome P450 Reductases From Legumes for Heterologous Production of Triterpenoids in Transgenic Saccharomyces cerevisiae |
title_sort | comparative analysis of nadph-cytochrome p450 reductases from legumes for heterologous production of triterpenoids in transgenic saccharomyces cerevisiae |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8716914/ https://www.ncbi.nlm.nih.gov/pubmed/34975947 http://dx.doi.org/10.3389/fpls.2021.762546 |
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