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Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds

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Abstract

The substrate specificity of the acyl–acyl carrier protein (ACP) thioesterases significantly determines the type of fatty acids that are exported from plastids. Thus, designing acyl-ACP thioesterases with different substrate specificities or kinetic properties would be of interest for plant lipid biotechnology to produce oils enriched in specialty fatty acids. In the present work, the FatA thioesterase from Helianthus annuus was used to test the impact of changes in the amino acids present in the binding pocket on substrate specificity and catalytic efficiency. Amongst all the mutated enzymes studied, Q215W was especially interesting as it had higher specificity towards saturated acyl-ACP substrates and higher catalytic efficiency compared to wild-type H. annuus FatA. Null, wild type and high-efficiency alleles were transiently expressed in tobacco leaves to check their effect on lipid biosynthesis. Expression of active FatA thioesterases altered the composition of leaf triacylglycerols but did not alter total lipid content. However, the expression of the wild type and the high-efficiency alleles in Arabidopsis thaliana transgenic seeds resulted in a strong reduction in oil content and an increase in total saturated fatty acid content. The role and influence of acyl-ACP thioesterases in plant metabolism and their possible applications in lipid biotechnology are discussed.

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Abbreviations

ACP:

Acyl carrier protein

CoA:

Coenzyme A

DAG:

Diacylglycerol

IMAC:

Immobilized metal affinity chromatography

TAG:

Triacylglycerides

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Acknowledgments

We are grateful to Rosario Sánchez and Valeria Gazda for their technical assistance. This work was supported by the MINECO and FEDER (Project AGL2011-23187).

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Authors and Affiliations

  1. Instituto de la Grasa, CSIC, Av. Padre García Tejero, 4, 41012, Sevilla, Spain

    Antonio Javier Moreno-Pérez, Mónica Venegas-Calerón, Joaquin J. Salas, Rafael Garcés & Enrique Martínez-Force

  2. Department of Biology, Centre for Novel Agricultural Products, Heslington, York, YO10 5DD, UK

    Fabián E. Vaistij, Tony R. Larson & Ian A. Graham

Authors
  1. Antonio Javier Moreno-Pérez
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  2. Mónica Venegas-Calerón
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  3. Fabián E. Vaistij
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  4. Joaquin J. Salas
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  5. Tony R. Larson
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  6. Rafael Garcés
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  7. Ian A. Graham
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  8. Enrique Martínez-Force
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Correspondence to Enrique Martínez-Force.

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Moreno-Pérez, A.J., Venegas-Calerón, M., Vaistij, F.E. et al. Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds. Planta 239, 667–677 (2014). https://doi.org/10.1007/s00425-013-2003-0

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