Abstract
As the world population grows, the demand for food increases. Although vegetable oils provide an affordable and rich source of energy, the supply of vegetable oils available for human consumption is limited by the “fuel vs food” debate. To increase the nutritional value of vegetable oil, metabolic engineering may be used to produce oil crops of desirable fatty acid composition. We have isolated and characterized β-ketoacyl ACP-synthase II (KASII) cDNA from a high-oleic acid palm, Jessenia bataua. Jessenia KASII (JbKASII) encodes a 488-amino acid polypeptide that possesses conserved domains that are necessary for condensing activities. When overexpressed in E. coli, recombinant His-tagged JbKASII was insoluble and non-functional. However, Arabidopsis plants expressing GFP-JbKASII fusions had elevated levels of arachidic acid (C20:0) and erucic acid (C22:1) at the expense of stearic acid (C18:0) and oleic acid (C18:1). Furthermore, JbKASII failed to complement the Arabidopsis KASII mutant, fab1-2. This suggests that the substrate specificity of JbKASII is similar to that of ketoacyl-CoA synthase (KCS), which preferentially elongates stearic and oleic acids, and not palmitic acid. Our results suggest that the KCS-like JbKASII may elongate C18:0 and C18:1 to yield C20:0 and C22:1, respectively. JbKASII may, therefore, be an interesting candidate gene for promoting the production of very long chain fatty acids in transgenic oil crops.
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Acknowledgments
We thank the Director-General of the Malaysian Palm Oil Board for permitting to publish this article, Rajanaidu a/l Nookiah for providing the Jessenia samples, and Mahadzir b. Jaafar for technical assistance with the gas chromatography analysis of fatty acids.
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An erratum to this article is available at http://dx.doi.org/10.1007/s12033-010-9361-7.
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12033_2010_9350_MOESM1_ESM.doc
Table S1 - Predicted cellular localizations of plant KASII homologs. The amino acid sequences of KASII homologs were subjected to motif analysis using PSORT (http://psort.hgc.jp/), and cellular localizations were predicted. +, likely cellular localization; UD, undetermined. The GenBank accession numbers of the KASII homolgs are listed after the species names. (DOC 33 kb)
12033_2010_9350_MOESM2_ESM.doc
Figure S1 - ClustalW multiple sequence alignment of plant Ketoacyl-CoA Synthases and JbKASII. Amino acid sequences of KCS homologs from Camellia oleifera (ACQ41892), Arabidopsis thaliana (AAF02814, Q38860), Lunaria annua (ACJ6177), and Brassica oleracea (AAC25112) were subjected to ClustalW multiple sequence alignment. KCS homologs showed significant sequence identity from the 180th amino acid to the 530th amino acid in the chalcone and stilbene synthase domain. There is little sequence similarity between JbKASII and the KCS homologs. (DOC 99 kb)
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Teh, OK., Ramli, U.S. Characterization of a KCS-like KASII from Jessenia bataua that Elongates Saturated and Monounsaturated Stearic Acids in Arabidopsis thaliana . Mol Biotechnol 48, 97–108 (2011). https://doi.org/10.1007/s12033-010-9350-x
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DOI: https://doi.org/10.1007/s12033-010-9350-x