Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering

A - Papers appearing in refereed journals

Ruiz-Lopez, N., Broughton, R., Usher, S. L., Salas, J. J., Haslam, R. P., Napier, J. A. and Beaudoin, F. 2017. Tailoring the composition of novel wax esters in the seeds of transgenic Camelina sativa through systematic metabolic engineering. Plant Biotechnology Journal. 15 (7), pp. 837-849. https://doi.org/10.1111/pbi.12679

AuthorsRuiz-Lopez, N., Broughton, R., Usher, S. L., Salas, J. J., Haslam, R. P., Napier, J. A. and Beaudoin, F.
Abstract

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.

Keywordsmetabolic engineering; seed lipids; Camelina; wax esters
Year of Publication2017
JournalPlant Biotechnology Journal
Journal citation15 (7), pp. 837-849
Digital Object Identifier (DOI)https://doi.org/10.1111/pbi.12679
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeDesigning Seeds for Nutrition and Health (DS)
Engineering oilseeds to synthesise designer wax esters
Renewable Industrial Products from Rapeseed: RIPR
Industrial crops producing added value oils for novel chemicals: ICON
The production of docosahexaenoic acid (DHA) in transgenic plants: a sustainable source of omega-3 fish oils
IDG/Manipulating epicuticular wax structure and composition to improve photosynthesis and increase the yield of Brassica oilseed crops
Publisher's version
Copyright license
CC BY
Output statusPublished
Publication dates
Online19 Dec 2016
Publication process dates
Accepted09 Dec 2016
PublisherWiley
ISSN1467-7644

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