Enzymatic characterisation of high-palmitic acid sunflower ( Helianthus annuus L.) mutants

Martı́nez-Force, Enrique; Álvarez-Ortega, Rosario; Garcés, Rafael

doi:10.1007/s004250050514

Cited by 31 publications

(27 citation statements)

References 32 publications

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“…The values for BnMCAT activity in crude extracts were comparable to those found in spinach and sunflower crude extracts [7] [13] [14]. The relative activity of BnMCAT was similar at different embryo stages, with little variation, and a higher relative BnMCAT activity was found in embryos than leaves.…”

Section: Enzymatic Activity Of Bnmcat In Rapeseedsupporting

confidence: 57%

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Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in <i>Brassica napus</i> Cultivars with Different Oil Content

Qu¹,

Jiang

Zhang³

2014

AJPS

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The full-length genomic DNA of MCAT (Malonyl-CoA:acyl carrier protein transacylase) in Brassica napus was cloned. BnMCAT shares very high identity with AtMCAT in gene sequence and gene structure. A multiple alignment of the protein sequence showed that BnMCAT shares high identity with other MCATs from E. coli and plants. BnMCAT was expressed in all tissues, such as roots, stems, leaves, flowers, and seeds, and no significant differences in the expression level were found in different embryo stages after pollination. According to an in vitro relative activity analysis, purified recombinant BnMCAT expressed in E. coli had transacylase activity. Although the relative activities of BnMCAT in crude extracts isolated from different staged embryos were similar and showed little variation, a higher relative activity was found in a crude extract isolated from embryos in comparison to leaves. Different relative activities of BnMCAT in crude extracts isolated from cultivars with different oil content were also found, suggesting that the activity of BnMCAT might be a decisive factor for a high oil content. Together, these results showed that BnMCAT is an important enzyme in the FAS system and indicate that BnMCAT might be a new target enzyme for future crop improvement through genetic engineering.

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Section: Enzymatic Activity Of Bnmcat In Rapeseedsupporting

confidence: 57%

“…The activity of malonyl-CoA:ACP transacylase was assayed in a final volume of 150 μl with 120 μl of crude extract using a modification of the method described by Martínez-Force et al [7] and Kremer et al [8]. The crude extract was omitted from control assays.…”

Section: Assay Of Malonyl-coa:acp Transacylase In Vitromentioning

confidence: 99%

Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in <i>Brassica napus</i> Cultivars with Different Oil Content

Qu¹,

Jiang

Zhang³

2014

AJPS

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“…Palmitoleic acid, 16:1 is not usually found in the lipid fraction of most plant seeds, including sunflower. Nevertheless, we tested this acyl-ACP here because appreciable amounts are found in high-palmitic acid sunflower mutants, up to 12.3% (Martı´nez-Force et al 1999;Salas et al 2004). Due to its stability and low melting point, this fatty acid is of interest in oleochemistry.…”

Section: Expression Of Sunflower Fata1 Thioesterase In Escherichia Colimentioning

confidence: 99%

Cloning, characterization and structural model of a FatA-type thioesterase from sunflower seeds (Helianthus annuus L.)

Serrano-Vega

Garcés²,

Martı́nez-Force³

2005

Planta

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The substrate specificity of acyl-acyl carrier protein (ACP) thioesterases (EC 3.1.2.14) determines the fatty acids available for the biosynthesis of storage and membrane lipids in seeds. In order to determine the mechanisms involved in the biosynthesis of fatty acids in sunflower seeds (Helianthus annuus L.), we isolated, cloned and sequenced a cDNA clone of acyl-ACP thioesterase from developing sunflower seeds, HaFatA1. Through the heterologous expression of HaFatA1 in Escherichia coli we have purified and characterized this enzyme, showing that sunflower HaFatA1 cDNA encodes a functional thioesterase with preference for monounsaturated acyl-ACPs. The HaFatA1 thioesterase was most efficient (kcat/K(m)) in catalyzing oleoyl-ACP, both in vivo and in vitro. By comparing this sequence with those obtained from public databases, we constructed a phylogenetic tree that included FatA and FatB thioesterases, as well as related prokaryotic proteins. The phylogenetic relationships support the endosymbiotic theory of the origin of eukaryotic cells and the suggestion that eubacteria from the delta-subdivision were the guest cells in the symbiosis with archaea. These prokaryotic proteins are more homologous to plant FatB, suggesting that the ancient thioesterases were more similar to FatB. Finally, using the available structure prediction methods, a 3D model of plant acyl-ACP thioesterases is proposed that reflects the combined data from direct mutagenesis and chimera studies. In addition, the model was tested by mutating the residues proposed to interact with the ACP protein in the FatA thioesterase by site-directed mutagenesis. The results indicate that this region is involved in the stabilization of the substrate at the active site.

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“…Notably, some TAG containing palmitolinoleic acid were present, such as PPlPo and PPlL in groups 3 and 4. The proportion of palmitolinoleic acid was low (0.5%) and it was generated, as reported previously in high-palmitic mutants, by the desaturation of palmitoleic acid by ER-localized ODS [26]. Furthermore, n-7 fatty acids were present at both the sn-2 and sn-1,3 positions, as demonstrated by pancreatic lipase digestion of these TAG (Tab.…”

Section: Tag Species Composition Of Cas-37 Oilmentioning

confidence: 54%

Characterization of the glycerolipid composition of a high‐palmitoleic acid sunflower mutant

Salas

Moreno‐Pérez

Martı́nez-Force

et al. 2007

Euro J Lipid Sci & Tech

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The glycerolipid composition of a high-palmitoleic acid sunflower (Helianthus annuus L.) mutant accumulating up to 20% of n-7 fatty acids was studied. This line produces oil with a complex triacylglycerol (TAG) composition, containing species that have not been previously identified in sunflower. In this regard, palmitoleic acid was esterified in an unexpected way in the three positions of the TAG molecules. The polar glycerolipid composition of the mutant was also studied, in order to identify and quantify the changes in membrane lipids imposed by the sunflower enzymatic machinery during the accumulation of the unusual n-7 fatty acids. The high-palmitoleic mutant accumulated important quantities of n-7 fatty acids in the polar lipid fraction, especially in the phosphatidylcholine lipid class. However, the total polar lipid content of these lines was not affected. On the other hand, the mutations responsible for the n-7 lipid accumulation induced an important decrease in the oil yield of the new mutant.

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Enzymatic characterisation of high-palmitic acid sunflower ( Helianthus annuus L.) mutants

Cited by 31 publications

References 32 publications

Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in <i>Brassica napus</i> Cultivars with Different Oil Content

Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in <i>Brassica napus</i> Cultivars with Different Oil Content

Cloning, characterization and structural model of a FatA-type thioesterase from sunflower seeds (Helianthus annuus L.)

Characterization of the glycerolipid composition of a high‐palmitoleic acid sunflower mutant

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Enzymatic characterisation of high-palmitic acid sunflower ( Helianthus annuus L.) mutants

Cited by 31 publications

References 32 publications

Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in &lt;i&gt;Brassica napus&lt;/i&gt; Cultivars with Different Oil Content

Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in &lt;i&gt;Brassica napus&lt;/i&gt; Cultivars with Different Oil Content

Cloning, characterization and structural model of a FatA-type thioesterase from sunflower seeds (Helianthus annuus L.)

Characterization of the glycerolipid composition of a high‐palmitoleic acid sunflower mutant

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Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in <i>Brassica napus</i> Cultivars with Different Oil Content

Cloning and Enzymatic Activity Analysis of the Malonyl-CoA:acyl Carrier Protein Transacylase in <i>Brassica napus</i> Cultivars with Different Oil Content