Abstract

Crambe is an oil crop suitable for industrial purposes due to the high content of erucic acid (22:1) in the seed oil. The final acylation of diacylglycerols (DAG) with acyl-CoA in the production of triacylglycerols (oil) is catalyzed by acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymes. We identified eight forms of DGATs in crambe and characterized them in microsomal preparations of yeast expressing the enzymes using various acyl-CoAs and both di-6:0-DAG and long-chain DAG species as acyl acceptors. All DGATs accepted 22:1-CoA when using di-6:0-DAG as acyl acceptor. When di-22:1-DAG was the acyl acceptor, the DGAT1 type of enzyme utilized 22:1-CoA at a much-reduced rate compared to assays with sn-1-22:1-sn-2-18:1(oleoyl)-DAG, the most frequently available DAG precursor in crambe seeds. None of the DGAT2 enzymes was able to acylate di-22:1-DAG. Our results indicate that formation of trierucin by crambe DGATs is a limiting step for further increasing the levels of 22:1 in the previously developed transgenic crambe lines due to their poor abilities to acylate di-22:1-DAG. We also show that the acyl-CoA specificities and the enzymatic activities are highly influenced by the fatty acid composition of the DAG acyl acceptor. This finding implies that the use of artificial acyl acceptors (e.g. di-6:0-DAG) may not always reflect the actual acyl-CoA specificities of DGATs in planta. The relevance of the here reported pronounced specificities for specific DAG species exerted by DGAT enzymes is discussed in the context of the findings of DAG pools of distinct catalytic origin in triacylglycerol biosynthesis in the seed oil.

Keywords

crambe; DGAT; DAG; TAG; Kennedy pathway; FA-composition

Published in

Frontiers in Plant Science
2019, Volume: 10, article number: 1442
Publisher: FRONTIERS MEDIA SA

SLU Authors

• Jeppsson, Simon

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Carlsson, Anders

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Zhu, Li-Hua

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Stymne, Sten

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Lager, Ida

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Biocatalysis and Enzyme Technology


Publication identifier

DOI: https://doi.org/10.3389/fpls.2019.01442

Permanent link to this page (URI)

https://res.slu.se/id/publ/103065