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Molecular cloning and characterization of a KCS gene from Cardamine graeca and its heterologous expression in Brassica oilseeds to engineer high nervonic acid oils for potential medical and industrial use.
Plant Biotechnol J. 2009 Dec; 7(9):925-38.PB

Abstract

Nervonic acid 24:1 Delta15 (cis-tetracos-15-enoic acid) is a very long-chain monounsaturated fatty acid and exists in nature as an elongation product of oleic acid. There is an increasing interest in production of high nervonic acid oils for pharmaceutical, nutraceutical and industrial applications. Using a polymerase chain reaction approach, we have isolated a gene from Cardamine graeca L., which encodes a 3-ketoacyl-CoA synthase (KCS), the first component of the elongation complex involved in synthesis of nervonic acid. Expression of the Cardamine KCS in yeast resulted in biosynthesis of nervonic acid, which is not normally present in yeast cells. We transformed Arabidopsis and Brassica carinata with the Cardamine KCS under the control of the seed-specific promoter, napin. The T(3) generations of transgenic Arabidopsis and B. carinata plants expressing the Cardamine KCS showed that seed-specific expression resulted in relatively large comparative increases in nervonic acid proportions in Arabidopsis seed oil, and 15-fold increase in nervonic acid proportions in B. carinata seed oil. The highest nervonic acid level in transgenic B. carinata lines reached 44%, with only 6% of residual erucic acid. In contrast, similar transgenic expression of the Cardamine KCS in high erucic B. napus resulted in 30% nervonic acid but with 20% residual erucic acid. Experiments using the Lunaria KCS gene gave results similar to the latter. In both cases, the erucic acid content is too high for human or animal consumption. Thus, the Cardamine KCS: B. carinata high nervonic/highly reduced erucic transgenic seed oils will be the most suitable for testing in pharmaceutical/nutraceutical applications to improve human and animal health.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Taylor DC
National Research Council of Canada, Plant Biotechnology Institute, Saskatoon, SK, Canada. david.taylor@nrc-cnrc.gc.ca
Francis T
No affiliation info available
Guo Y
No affiliation info available
Brost JM
No affiliation info available
Katavic V
No affiliation info available
Mietkiewska E
No affiliation info available
Michael Giblin E
No affiliation info available
Lozinsky S
No affiliation info available
Hoffman T
No affiliation info available

MeSH

3-Oxoacyl-(Acyl-Carrier-Protein) SynthaseBrassicaCardamineCloning, MolecularDNA, PlantFatty Acids, MonounsaturatedGene Expression Regulation, PlantGenes, PlantPlant OilsPlants, Genetically ModifiedTransformation, Genetic

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19843251

Citation

Taylor, David C., et al. "Molecular Cloning and Characterization of a KCS Gene From Cardamine Graeca and Its Heterologous Expression in Brassica Oilseeds to Engineer High Nervonic Acid Oils for Potential Medical and Industrial Use." Plant Biotechnology Journal, vol. 7, no. 9, 2009, pp. 925-38.
Taylor DC, Francis T, Guo Y, et al. Molecular cloning and characterization of a KCS gene from Cardamine graeca and its heterologous expression in Brassica oilseeds to engineer high nervonic acid oils for potential medical and industrial use. Plant Biotechnol J. 2009;7(9):925-38.
Taylor, D. C., Francis, T., Guo, Y., Brost, J. M., Katavic, V., Mietkiewska, E., Michael Giblin, E., Lozinsky, S., & Hoffman, T. (2009). Molecular cloning and characterization of a KCS gene from Cardamine graeca and its heterologous expression in Brassica oilseeds to engineer high nervonic acid oils for potential medical and industrial use. Plant Biotechnology Journal, 7(9), 925-38. https://doi.org/10.1111/j.1467-7652.2009.00454.x
Taylor DC, et al. Molecular Cloning and Characterization of a KCS Gene From Cardamine Graeca and Its Heterologous Expression in Brassica Oilseeds to Engineer High Nervonic Acid Oils for Potential Medical and Industrial Use. Plant Biotechnol J. 2009;7(9):925-38. PubMed PMID: 19843251.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular cloning and characterization of a KCS gene from Cardamine graeca and its heterologous expression in Brassica oilseeds to engineer high nervonic acid oils for potential medical and industrial use. AU - Taylor,David C, AU - Francis,Tammy, AU - Guo,Yiming, AU - Brost,Jennifer M, AU - Katavic,Vesna, AU - Mietkiewska,Elzbieta, AU - Michael Giblin,E, AU - Lozinsky,Sharla, AU - Hoffman,Travis, Y1 - 2009/10/13/ PY - 2009/10/22/entrez PY - 2009/10/22/pubmed PY - 2010/1/15/medline SP - 925 EP - 38 JF - Plant biotechnology journal JO - Plant Biotechnol J VL - 7 IS - 9 N2 - Nervonic acid 24:1 Delta15 (cis-tetracos-15-enoic acid) is a very long-chain monounsaturated fatty acid and exists in nature as an elongation product of oleic acid. There is an increasing interest in production of high nervonic acid oils for pharmaceutical, nutraceutical and industrial applications. Using a polymerase chain reaction approach, we have isolated a gene from Cardamine graeca L., which encodes a 3-ketoacyl-CoA synthase (KCS), the first component of the elongation complex involved in synthesis of nervonic acid. Expression of the Cardamine KCS in yeast resulted in biosynthesis of nervonic acid, which is not normally present in yeast cells. We transformed Arabidopsis and Brassica carinata with the Cardamine KCS under the control of the seed-specific promoter, napin. The T(3) generations of transgenic Arabidopsis and B. carinata plants expressing the Cardamine KCS showed that seed-specific expression resulted in relatively large comparative increases in nervonic acid proportions in Arabidopsis seed oil, and 15-fold increase in nervonic acid proportions in B. carinata seed oil. The highest nervonic acid level in transgenic B. carinata lines reached 44%, with only 6% of residual erucic acid. In contrast, similar transgenic expression of the Cardamine KCS in high erucic B. napus resulted in 30% nervonic acid but with 20% residual erucic acid. Experiments using the Lunaria KCS gene gave results similar to the latter. In both cases, the erucic acid content is too high for human or animal consumption. Thus, the Cardamine KCS: B. carinata high nervonic/highly reduced erucic transgenic seed oils will be the most suitable for testing in pharmaceutical/nutraceutical applications to improve human and animal health. SN - 1467-7652 UR - https://www.unboundmedicine.com/medline/citation/19843251/Molecular_cloning_and_characterization_of_a_KCS_gene_from_Cardamine_graeca_and_its_heterologous_expression_in_Brassica_oilseeds_to_engineer_high_nervonic_acid_oils_for_potential_medical_and_industrial_use_ L2 - https://doi.org/10.1111/j.1467-7652.2009.00454.x DB - PRIME DP - Unbound Medicine ER -