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...

Descripción completa

Detalles Bibliográficos
Autores principales: Ruiz‐Lopez, Noemi, Broughton, Richard, Usher, Sarah, Salas, Joaquin J., Haslam, Richard P., Napier, Johnathan A., Beaudoin, Frédéric
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