Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering
The functional characterization of wax biosynthetic enzymes in transgenic plants has opened the possibility of producing tailored wax esters (WEs) in the seeds of a suitable host crop. In this study, in addition to systematically evaluating a panel of WE biosynthetic activities, we have also modulat...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466440/ https://www.ncbi.nlm.nih.gov/pubmed/27990737 http://dx.doi.org/10.1111/pbi.12679 |
_version_ | 1783243100599091200 |
---|---|
author | Ruiz‐Lopez, Noemi Broughton, Richard Usher, Sarah Salas, Joaquin J. Haslam, Richard P. Napier, Johnathan A. Beaudoin, Frédéric |
author_facet | Ruiz‐Lopez, Noemi Broughton, Richard Usher, Sarah Salas, Joaquin J. Haslam, Richard P. Napier, Johnathan A. Beaudoin, Frédéric |
author_sort | Ruiz‐Lopez, Noemi |
collection | PubMed |
description | The functional characterization of wax biosynthetic enzymes in transgenic plants has opened the possibility of producing tailored wax esters (WEs) in the seeds of a suitable host crop. In this study, in addition to systematically evaluating a panel of WE biosynthetic activities, we have also modulated the acyl‐CoA substrate pool, through the co‐expression of acyl‐ACP thioesterases, to direct the accumulation of medium‐chain fatty acids. Using this combinatorial approach, we determined the additive contribution of both the varied acyl‐CoA pool and biosynthetic enzyme substrate specificity to the accumulation of non‐native WEs in the seeds of transgenic Camelina plants. A total of fourteen constructs were prepared containing selected FAR and WS genes in combination with an acyl‐ACP thioesterase. All enzyme combinations led to the successful production of wax esters, of differing compositions. The impact of acyl‐CoA thioesterase expression on wax ester accumulation varied depending on the substrate specificity of the WS. Hence, co‐expression of acyl‐ACP thioesterases with Marinobacter hydrocarbonoclasticus WS and Marinobacter aquaeolei FAR resulted in the production of WEs with reduced chain lengths, whereas the co‐expression of the same acyl‐ACP thioesterases in combination with Mus musculus WS and M. aquaeolei FAR had little impact on the overall final wax composition. This was despite substantial remodelling of the acyl‐CoA pool, suggesting that these substrates were not efficiently incorporated into WEs. These results indicate that modification of the substrate pool requires careful selection of the WS and FAR activities for the successful high accumulation of these novel wax ester species in Camelina seeds. |
format | Online Article Text |
id | pubmed-5466440 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-54664402017-06-21 Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering Ruiz‐Lopez, Noemi Broughton, Richard Usher, Sarah Salas, Joaquin J. Haslam, Richard P. Napier, Johnathan A. Beaudoin, Frédéric Plant Biotechnol J Research Articles The functional characterization of wax biosynthetic enzymes in transgenic plants has opened the possibility of producing tailored wax esters (WEs) in the seeds of a suitable host crop. In this study, in addition to systematically evaluating a panel of WE biosynthetic activities, we have also modulated the acyl‐CoA substrate pool, through the co‐expression of acyl‐ACP thioesterases, to direct the accumulation of medium‐chain fatty acids. Using this combinatorial approach, we determined the additive contribution of both the varied acyl‐CoA pool and biosynthetic enzyme substrate specificity to the accumulation of non‐native WEs in the seeds of transgenic Camelina plants. A total of fourteen constructs were prepared containing selected FAR and WS genes in combination with an acyl‐ACP thioesterase. All enzyme combinations led to the successful production of wax esters, of differing compositions. The impact of acyl‐CoA thioesterase expression on wax ester accumulation varied depending on the substrate specificity of the WS. Hence, co‐expression of acyl‐ACP thioesterases with Marinobacter hydrocarbonoclasticus WS and Marinobacter aquaeolei FAR resulted in the production of WEs with reduced chain lengths, whereas the co‐expression of the same acyl‐ACP thioesterases in combination with Mus musculus WS and M. aquaeolei FAR had little impact on the overall final wax composition. This was despite substantial remodelling of the acyl‐CoA pool, suggesting that these substrates were not efficiently incorporated into WEs. These results indicate that modification of the substrate pool requires careful selection of the WS and FAR activities for the successful high accumulation of these novel wax ester species in Camelina seeds. John Wiley and Sons Inc. 2017-02-02 2017-07 /pmc/articles/PMC5466440/ /pubmed/27990737 http://dx.doi.org/10.1111/pbi.12679 Text en © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Ruiz‐Lopez, Noemi Broughton, Richard Usher, Sarah Salas, Joaquin J. Haslam, Richard P. Napier, Johnathan A. Beaudoin, Frédéric Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering |
title | Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering |
title_full | Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering |
title_fullStr | Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering |
title_full_unstemmed | Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering |
title_short | Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering |
title_sort | tailoring the composition of novel wax esters in the seeds of transgenic camelina sativa through systematic metabolic engineering |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466440/ https://www.ncbi.nlm.nih.gov/pubmed/27990737 http://dx.doi.org/10.1111/pbi.12679 |
work_keys_str_mv | AT ruizlopeznoemi tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering AT broughtonrichard tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering AT ushersarah tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering AT salasjoaquinj tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering AT haslamrichardp tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering AT napierjohnathana tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering AT beaudoinfrederic tailoringthecompositionofnovelwaxestersintheseedsoftransgeniccamelinasativathroughsystematicmetabolicengineering |