Abstract

BackgroundWax esters are highly hydrophobic neutral lipids that are major constituents of the cutin and suberin layer. Moreover they have favorable properties as a commodity for industrial applications. Through transgenic expression of wax ester biosynthetic genes in oilseed crops, it is possible to achieve high level accumulation of defined wax ester compositions within the seed oil to provide a sustainable source for such high value lipids. The fatty alcohol moiety of the wax esters is formed from plant-endogenous acyl-CoAs by the action of fatty acyl reductases (FAR). In a second step the fatty alcohol is condensed with acyl-CoA by a wax synthase (WS) to form a wax ester. In order to evaluate the specificity of wax ester biosynthesis, analytical methods are needed that provide detailed wax ester profiles from complex lipid extracts.ResultsWe present a direct infusion ESI-tandem MS method that allows the semi-quantitative determination of wax ester compositions from complex lipid mixtures covering 784 even chain molecular species. The definition of calibration prototype groups that combine wax esters according to their fragmentation behavior enables fast quantitative analysis by applying multiple reaction monitoring. This provides a tool to analyze wax layer composition or determine whether seeds accumulate a desired wax ester profile. Besides the profiling method, we provide general information on wax ester analysis by the systematic definition of wax ester prototypes according to their collision-induced dissociation spectra. We applied the developed method for wax ester profiling of the well characterized jojoba seed oil and compared the profile with wax ester-accumulating Arabidopsis thaliana expressing the wax ester biosynthetic genes MaFAR and ScWS.ConclusionsWe developed a fast profiling method for wax ester analysis on the molecular species level. This method is suitable to screen large numbers of transgenic plants as well as other wax ester samples like cuticular lipid extracts to gain an overview on the molecular species composition. We confirm previous results from APCI-MS and GC-MS analysis, which showed that fragmentation patterns are highly dependent on the double bond distribution between the fatty alcohol and the fatty acid part of the wax ester.

Highlights

  • Wax esters are highly hydrophobic neutral lipids that are major constituents of the cutin and suberin layer

  • To evaluate the wax ester composition of seed oil obtained from transgenic plants, there is the need for analytical methods that enable quantitative wax ester profiling on the molecular species level

  • Wax ester profiling is taken as a mean to evaluate the quality of edible oils, especially olive oil, that contains residual amounts of wax esters residing from the seed or fruit coat [32,33]

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Summary

Introduction

Wax esters are highly hydrophobic neutral lipids that are major constituents of the cutin and suberin layer They have favorable properties as a commodity for industrial applications. Through transgenic expression of wax ester biosynthetic genes in oilseed crops, it is possible to achieve high level accumulation of defined wax ester compositions within the seed oil to provide a sustainable source for such high value lipids. To evaluate the wax ester composition of seed oil obtained from transgenic plants, there is the need for analytical methods that enable quantitative wax ester profiling on the molecular species level. Wax ester profiling could be highly useful for the elucidation of specific compositions of epicuticular waxes from different plant sources. Wax ester profiling is taken as a mean to evaluate the quality of edible oils, especially olive oil, that contains residual amounts of wax esters residing from the seed or fruit coat [32,33]

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