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Acyl carrier proteins from sunflower (Helianthus annuus L.) seeds and their influence on FatA and FatB acyl-ACP thioesterase activities


Main conclusion

The kinetics of acyl-ACP thioesterases from sunflower importantly changed when endogenous ACPs were used. Sunflower FatB was much more specific towards saturated acyl-ACPs when assayed with them.

Acyl carrier proteins (ACPs) are small (~9 kDa), soluble, acidic proteins involved in fatty acid synthesis in plants and bacteria. ACPs bind to fatty acids through a thioester bond, generating the acyl-ACP lipoproteins that are substrates for fatty acid synthase (FAS) complexes, and that are required for fatty acid chain elongation, acting as important intermediates in de novo fatty acid synthesis in plants. Plants, usually express several ACP isoforms with distinct functionalities. We report here the cloning of three ACPs from developing sunflower seeds: HaACP1, HaACP2, and HaACP3. These proteins were plastidial ACPs expressed strongly in seeds, and as such they are probably involved in the synthesis of sunflower oil. The recombinant sunflower ACPs were expressed in bacteria but they were lethal to the prokaryote host. Thus, they were finally produced using the GST gene fusion system, which allowed the apo-enzyme to be produced and later activated to the holo form. Radiolabelled acyl-ACPs from the newly cloned holo-ACP forms were also synthesized and used to characterize the activity of recombinant sunflower FatA and FatB thioesterases, important enzymes in plant fatty acids synthesis. The activity of these enzymes changed significantly when the endogenous ACPs were used. Thus, FatA importantly increased its activity levels, whereas FatB displayed a different specificity profile, with much high activity levels towards saturated acyl-CoA derivatives. All these data pointed to an important influence of the ACP moieties on the activity of enzymes involved in lipid synthesis.

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Acyl carrier protein


Days after flowering




Fatty acid synthases


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  1. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res 25:3389–3402

  2. Battey JF, Ohlrogge JB (1990) Evolutionary and tissue-specific control of expression of multiple acyl-carrier protein isoforms in plants and bacteria. Planta 180:352–360

  3. Beisson F, Koo AJK, Ruuska S, Schwender J, Pollard MR, Thelen JJ, Paddock T, Salas JJ, Savage L, Milcamps A, Mhaske VB, Cho Y, Ohlrogge JB (2003) Arabidopsis genes involved in acyl lipid metabolism. A 2003 census of the candidates, a study of the distribution of expressed sequence tags in organs, and a web-based database. Plant Physiol 132:681–697

  4. Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE (2000) The protein data bank. Nucl Acids Res 28:235–242

  5. Bonaventure G, Salas JJ, Pollard MR, Ohlrogge JB (2003) Disruption of the FATB gene in arabidopsis demonstrates an essential role of saturated fatty acids in plant growth. Plant Cell 15:1020–1033

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  7. Chen M, Han G, Dietrich CR, Dunn TM, Cahoon EB (2006) The essential nature of sphingolipids in plants as revealed by the functional identification and characterization of the Arabidopsis LCB1 subunit of serine palmitoyltransferase. Plant Cell 18:3576–3593

  8. Dörmann P, Voelker VA, Ohlrogge JB (1995) Cloning and expression in Escherichia coli of a novel thioesterase from arabidopsis thaliana specific for long-chain acyl-acyl carrier proteins. Arch Biochem Bioph 316:612–618

  9. Dörmann P, Voelker TA, Ohlrogge JB (2000) Accumulation of palmitate in arabidopsis mediated by the acyl-acyl carrier protein thioesterase FATB1. Plant Physiol 123:637–644

  10. Ghosh SK, Bhattacharjee A, Jha JK, Mondal AK, Maiti MK, Basu A, Ghosh D, Ghosh S, Sen SK (2007) Characterization and cloning of a stearoyl/oleoyl specific fatty acyl–acyl carrier protein thioesterase from the seeds of Madhuca longifolia (latifolia). Plant Physiol Biochem 45:887–897

  11. González-Thuillier I, Venegas-Calerón M, Garcés R, Wettstein-Knowles P, Martínez-Force E (2015) Sunflower (Helianthus annuus) fatty acid synthase complex: enoyl-(acyl carrier protein)-reductase genes. Planta 241:43–56

  12. Guex N, Peitsch MC (1997) SWISS-MODEL and the Swiss-Pdb Viewer: an environment for comparative protein modelling. Electrophoresis 18:2714–2723

  13. Hawkins DJ, Kridl JC (1998) Characterization of acyl-ACP thioesterases of mangosteen (Garcinia mangostana) seed and high levels of stearate production in transgenic canola. Plant J 13:743–752

  14. Hiltunen JK, Schonauer MS, Autio KJ, Mittelmeier TM, Kastaniotis AJ, Dieckmann CL (2009) Mitochondrial fatty acid synthesis type II: more than just fatty acids. J Biol Chem 284:9011–9015

  15. Holak TA, Kearsley SK, Kim Y, Prestegard JH (1988) Three-dimensional structure of acyl carrier protein determined by NMR pseudoenergy and distance geometry calculations. Biochemistry 27:6135–6142

  16. Jones A, Davies HM, Voelker TA (1995) Palmitoyl-acyl carrier protein (ACP) thioesterase and the evolutionary origin of plant acyl-ACP thioesterases. Plant Cell 7:359–371

  17. Kim Y, Kovrigin EL, Eletr Z (2006) NMR studies of Escherichia coli acyl carrier protein: dynamic and structural differences of the apo- and holo-forms. Biochem Bioph Res Co 341:776–783

  18. Kuo TM, Ohlrogge JB (1984) Acylation of plant acyl carrier proteins by acyl-acyl carrier protein synthetase from Escherichia coli. Arch Biochem Bioph 230:110–116

  19. Lambalot RH, Walsh CT (1995) Cloning, overproduction, and characterization of the Escherichia coli holo-acyl carrier protein synthase. J Biol Chem 270:24658–24661

  20. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and clustal X version 2.0. Bioinformatics 23:2947–2948

  21. Li Q, Khosla C, Puglisi JD, Liu CW (2003) Solution structure and backbone dynamics of the holo form of the frenolicin acyl carrier protein. Biochemistry 42:4648–4657

  22. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2 − ΔΔCT method. Methods 25:402–408

  23. Moreno-Pérez AJ, Sánchez-García A, Salas JJ, Garcés R, Martínez-Force E (2011) Acyl-ACP thioesterases from macadamia (Macadamia tetraphylla) nuts: cloning, characterization and their impact on oil composition. Plant Physiol Biochem 49:82–87

  24. Moreno-Pérez AJ, Venegas-Calerón M, Vaistij FE, Salas JJ, Larson TR, Garcés R, Graham IA, Martínez-Force E (2012) Reduced expression of FatA thioesterases in Arabidopsis affects the oil content and fatty acid composition of the seeds. Planta 235:629–639

  25. Ohlrogge JB, Jaworski JG (1997) Regulation of fatty acid synthesis. Annu Rev Plant Biol 48:109–136

  26. Pérez-Vich B, Garcés R, Fernández-Martínez JM (1999) Genetic control of high stearic acid content in the seed oil of the sunflower mutant CAS-3. Theor Appl Genet 99:663–669

  27. Pleite R, Martínez-Force E, Garcés R (2006) Increase of the stearic acid content in high-oleic sunflower (Helianthus annuus) Seeds. J Agric Food Chem 54:9383–9388

  28. Pollard MR, Anderson L, Fan C, Hawkins DJ, Davies MH (1991) A specific acyl-ACP thioesterase implicated in medium-chain fatty acid production in immature cotyledons of Umbellularia californica. Arch Biochem Bioph 284:306–312

  29. Rawlings M, Cronan JE (1992) The gene encoding Escherichia coli acyl carrier protein lies within a cluster of fatty acid biosynthetic genes. J Biol Chem 267:5751–5754

  30. Rock CO, Garwin JL (1979) Preparative enzymatic synthesis and hydrophobic chromatography of acyl-acyl carrier protein. J Biol Chem 254:7123–7128

  31. Salas JJ, Ohlrogge JB (2002) Characterization of substrate specificity of plant FatA and FatB acyl-ACP thioesterases. Arch Biochem Biophys 403:25–34

  32. Sánchez-García A, Moreno-Pérez AJ, Muro-Pastor AM, Salas JJ, Garcés R, Martínez-Force E (2010) Acyl-ACP thioesterases from castor (Ricinus communis L.): an enzymatic system appropriate for high rates of oil synthesis and accumulation. Phytochemistry 71:860–869

  33. Serrano-Vega MJ, Venegas-Calerón M, Garcés R, Martínez-Force E (2003) Cloning and expression of fatty acids biosynthesis key enzymes from sunflower (Helianthus annuus L.) in Escherichia coli. J Chromatog B 786:221–228

  34. Serrano-Vega MJ, Garcés R, Martínez-Force E (2005) Cloning, characterization and structural model of a FatA-type thioesterase from sunflower seeds (Helianthus annuus L.). Planta 221:868–880

  35. Smith DB, Johnson KS (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67:31–40

  36. Suh MC, Schultz DJ, Ohlrogge JB (1999) Isoforms of acyl carrier protein involved in seed-specific fatty acid synthesis. Plant J 17:679–688

  37. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

  38. Thelen JJ, Ohlrogge JB (2002) Metabolic engineering of fatty acid biosynthesis in plants. Metab Eng 4:12–21

  39. Voelker T (1996) Plant Acyl-ACP Thioesterases: chain-length determining enzymes in plant fatty acid biosynthesis. Genetic Eng 18:111–133

  40. Wakil SJ (1989) Fatty acid synthase, a proficient multifunctional enzyme. Biochemistry 28:4523–4530

  41. White SW, Zheng J, Zhang Y-M, Rock CO (2005) The structural biology of type II fatty acid biosynthesis. Annu Rev Biochem 74:791–831

  42. Wu P-Z, Li J, Wei Q, Zeng L, Chen Y-P, Li MR, Jiang HW, Wu G-J (2009) Cloning and functional characterization of an acyl-acyl carrier protein thioesterase (JcFATB1) from Jatropha curcas. Tree Physiol 29:1299–1305

  43. Zornetzer GA, Fox BG, Markley JL (2006) Solution structures of spinach acyl carrier protein with decanoate and stearate. Biochemistry 45:5217–5227

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This work was funded by the “Ministerio de Economia y Competitividad” and FEDER (Project AGL2014-53537-R and JAE-CSIC to J.A. A-M).

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Correspondence to Joaquín J. Salas.

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Aznar-Moreno, J.A., Venegas-Calerón, M., Martínez-Force, E. et al. Acyl carrier proteins from sunflower (Helianthus annuus L.) seeds and their influence on FatA and FatB acyl-ACP thioesterase activities. Planta 244, 479–490 (2016).

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  • Acyl carrier protein
  • Sunflower, Helianthus annuus
  • Acyl-ACP thioesterase
  • Fatty acid synthesis