Transfer of the cytochrome P450-dependent dhurrin pathway from Sorghum bicolor into Nicotiana tabacum chloroplasts for light-driven synthesis

© The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Bibliographische Detailangaben
Veröffentlicht in:Journal of experimental botany. - 1985. - 67(2016), 8 vom: 15. Apr., Seite 2495-506
1. Verfasser: Gnanasekaran, Thiyagarajan (VerfasserIn)
Weitere Verfasser: Karcher, Daniel, Nielsen, Agnieszka Zygadlo, Martens, Helle Juel, Ruf, Stephanie, Kroop, Xenia, Olsen, Carl Erik, Motawie, Mohammed Saddik, Pribil, Mathias, Møller, Birger Lindberg, Bock, Ralph, Jensen, Poul Erik
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2016
Zugriff auf das übergeordnete Werk:Journal of experimental botany
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Cytochrome P450 chloroplast dhurrin pathway light-driven photosynthesis thylakoids. Glucosides Nitriles mehr... Protein Subunits Cytochrome P-450 Enzyme System 9035-51-2 dhurrin P5999IY65C
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100 1 |a Gnanasekaran, Thiyagarajan  |e verfasserin  |4 aut 
245 1 0 |a Transfer of the cytochrome P450-dependent dhurrin pathway from Sorghum bicolor into Nicotiana tabacum chloroplasts for light-driven synthesis 
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500 |a Date Revised 13.12.2023 
500 |a published: Print-Electronic 
500 |a CommentIn: J Exp Bot. 2016 Apr;67(8):2123-5. - PMID 27022183 
500 |a Citation Status MEDLINE 
520 |a © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology. 
520 |a Plant chloroplasts are light-driven cell factories that have great potential to act as a chassis for metabolic engineering applications. Using plant chloroplasts, we demonstrate how photosynthetic reducing power can drive a metabolic pathway to synthesise a bio-active natural product. For this purpose, we stably engineered the dhurrin pathway from Sorghum bicolor into the chloroplasts of Nicotiana tabacum (tobacco). Dhurrin is a cyanogenic glucoside and its synthesis from the amino acid tyrosine is catalysed by two membrane-bound cytochrome P450 enzymes (CYP79A1 and CYP71E1) and a soluble glucosyltransferase (UGT85B1), and is dependent on electron transfer from a P450 oxidoreductase. The entire pathway was introduced into the chloroplast by integrating CYP79A1, CYP71E1, and UGT85B1 into a neutral site of the N. tabacum chloroplast genome. The two P450s and the UGT85B1 were functional when expressed in the chloroplasts and converted endogenous tyrosine into dhurrin using electrons derived directly from the photosynthetic electron transport chain, without the need for the presence of an NADPH-dependent P450 oxidoreductase. The dhurrin produced in the engineered plants amounted to 0.1-0.2% of leaf dry weight compared to 6% in sorghum. The results obtained pave the way for plant P450s involved in the synthesis of economically important compounds to be engineered into the thylakoid membrane of chloroplasts, and demonstrate that their full catalytic cycle can be driven directly by photosynthesis-derived electrons 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Cytochrome P450 
650 4 |a chloroplast 
650 4 |a dhurrin pathway 
650 4 |a light-driven 
650 4 |a photosynthesis 
650 4 |a thylakoids. 
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650 7 |a Nitriles  |2 NLM 
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700 1 |a Karcher, Daniel  |e verfasserin  |4 aut 
700 1 |a Nielsen, Agnieszka Zygadlo  |e verfasserin  |4 aut 
700 1 |a Martens, Helle Juel  |e verfasserin  |4 aut 
700 1 |a Ruf, Stephanie  |e verfasserin  |4 aut 
700 1 |a Kroop, Xenia  |e verfasserin  |4 aut 
700 1 |a Olsen, Carl Erik  |e verfasserin  |4 aut 
700 1 |a Motawie, Mohammed Saddik  |e verfasserin  |4 aut 
700 1 |a Pribil, Mathias  |e verfasserin  |4 aut 
700 1 |a Møller, Birger Lindberg  |e verfasserin  |4 aut 
700 1 |a Bock, Ralph  |e verfasserin  |4 aut 
700 1 |a Jensen, Poul Erik  |e verfasserin  |4 aut 
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