Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana
Glucosinolates are natural products characteristic of the Brassicales order which include vegetables such as cabbages and the model plant Arabidopsis thaliana. Glucoraphanin is the major glucosinolate in broccoli and associated with the health-promoting effects of broccoli consumption. Towards our g...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2016-02-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fbioe.2016.00014/full |
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| author | Christoph eCrocoll Christoph eCrocoll Nadia eMirza Nadia eMirza Michael eReichelt Jonathan eGershenzon Barbara Ann Halkier Barbara Ann Halkier |
| author_facet | Christoph eCrocoll Christoph eCrocoll Nadia eMirza Nadia eMirza Michael eReichelt Jonathan eGershenzon Barbara Ann Halkier Barbara Ann Halkier |
| author_sort | Christoph eCrocoll |
| collection | DOAJ |
| description | Glucosinolates are natural products characteristic of the Brassicales order which include vegetables such as cabbages and the model plant Arabidopsis thaliana. Glucoraphanin is the major glucosinolate in broccoli and associated with the health-promoting effects of broccoli consumption. Towards our goal of creating a rich source of glucoraphanin for dietary supplements, we have previously reported the feasibility of engineering glucoraphanin in Nicotiana benthaminana through transient expression of glucoraphanin biosynthetic genes from Arabidopsis thaliana (Mikkelsen et al., 2010). As side-products, we obtained 5-8 fold higher levels of chain-elongated leucine-derived glucosinolates, not found in the native plant. Here, we investigated two different strategies to improve engineering of the methionine chain elongation part of the glucoraphanin pathway in N. benthamiana: 1) co-expression of the large subunit (LSU1) of the heterodimeric isopropylmalate isomerase, and 2) co-expression of BAT5 transporter for efficient transfer of intermediates across the chloroplast membrane. We succeeded in raising dihomo-methionine (DHM) levels to a maximum of 432 nmol*g-1 fresh weight which is equivalent to a 9-fold increase compared to the highest production of this intermediate previously reported (Mikkelsen et al., 2010). The increased DHM production without increasing leucine-derived side-product levels provides new metabolic engineering strategies for improved glucoraphanin production in a heterologous host. |
| first_indexed | 2024-12-22T09:33:00Z |
| format | Article |
| id | doaj.art-ac517e4f1efd45d4a0c8b741704d0627 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-22T09:33:00Z |
| publishDate | 2016-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-ac517e4f1efd45d4a0c8b741704d06272022-12-21T18:30:54ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852016-02-01410.3389/fbioe.2016.00014178767Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamianaChristoph eCrocoll0Christoph eCrocoll1Nadia eMirza2Nadia eMirza3Michael eReichelt4Jonathan eGershenzon5Barbara Ann Halkier6Barbara Ann Halkier7University of CopenhagenUniveristy of CopenhagenUniversity of CopenhagenUniveristy of CopenhagenMax Planck Institute for Chemical EcologyMax Planck Institute for Chemical EcologyUniversity of CopenhagenUniveristy of CopenhagenGlucosinolates are natural products characteristic of the Brassicales order which include vegetables such as cabbages and the model plant Arabidopsis thaliana. Glucoraphanin is the major glucosinolate in broccoli and associated with the health-promoting effects of broccoli consumption. Towards our goal of creating a rich source of glucoraphanin for dietary supplements, we have previously reported the feasibility of engineering glucoraphanin in Nicotiana benthaminana through transient expression of glucoraphanin biosynthetic genes from Arabidopsis thaliana (Mikkelsen et al., 2010). As side-products, we obtained 5-8 fold higher levels of chain-elongated leucine-derived glucosinolates, not found in the native plant. Here, we investigated two different strategies to improve engineering of the methionine chain elongation part of the glucoraphanin pathway in N. benthamiana: 1) co-expression of the large subunit (LSU1) of the heterodimeric isopropylmalate isomerase, and 2) co-expression of BAT5 transporter for efficient transfer of intermediates across the chloroplast membrane. We succeeded in raising dihomo-methionine (DHM) levels to a maximum of 432 nmol*g-1 fresh weight which is equivalent to a 9-fold increase compared to the highest production of this intermediate previously reported (Mikkelsen et al., 2010). The increased DHM production without increasing leucine-derived side-product levels provides new metabolic engineering strategies for improved glucoraphanin production in a heterologous host.http://journal.frontiersin.org/Journal/10.3389/fbioe.2016.00014/fullGlucosinolatesMetabolic EngineeringNicotiana benthamianaGlucoraphanindihomo-methionine |
| spellingShingle | Christoph eCrocoll Christoph eCrocoll Nadia eMirza Nadia eMirza Michael eReichelt Jonathan eGershenzon Barbara Ann Halkier Barbara Ann Halkier Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana Frontiers in Bioengineering and Biotechnology Glucosinolates Metabolic Engineering Nicotiana benthamiana Glucoraphanin dihomo-methionine |
| title | Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana |
| title_full | Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana |
| title_fullStr | Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana |
| title_full_unstemmed | Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana |
| title_short | Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana |
| title_sort | optimization of engineered production of the glucoraphanin precursor dihomo methionine in nicotiana benthamiana |
| topic | Glucosinolates Metabolic Engineering Nicotiana benthamiana Glucoraphanin dihomo-methionine |
| url | http://journal.frontiersin.org/Journal/10.3389/fbioe.2016.00014/full |
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