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Cloning and identification of a novel C18-Δ9 polyunsaturated fatty acid specific elongase gene from DHA-producing Isochrysis galbana H29

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Abstract

Isochrysis galbana, a marine prymnesiophyte microalga, is able to produce a high level of long chain polyunsaturated fatty acids such as docosahexaenoic acid (DHA, C22:6n-3). In this article, a novel gene (IgASE2) that encoded a C18-Δ9 polyunsaturase fatty acids specific (C18-Δ9-PUFAs-specific) elongase was isolated and characterized from DHA-rich microalga, I. galbana H29. A full-length cDNA of 1653 bp was cloned by rapidamplification of cDNA ends (RACE) PCR techniques. The IgASE2 contained a 786 bp ORF encoding a protein of 261 amino acids that shared 87% identity with the reported Δ9-elongase IgASE1, a 44 bp 5′ untranslated region and an 823 bp 3′ untranslated region. The function of IgASE2 was demonstrated by its heterologous expression in Saccharomyces cerevisiae. In S. cerevisiae, IgASE2 elongated linoleic acid (LA, C18:2n-6), α-linolenic (ALA, C18:3n-3) to eicosadienoic acid (EDA, C20:2n-6) and eicosatrienoic acid (ETrA, C20:3n-3). The conversion ratios of LA to EDA and ALA to ETrA were 60.47 and 58.36%, respectively. However, IgASE2 could not catalyze the elongation reactions of oleic acid (OA, C18:1n-9) and other fatty acids. These results confirmed that IgASE2 had C18-Δ9-PUFAs-specific elongase activity.

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Authors and Affiliations

  1. Key Laboratory of Molecular Microbiology and Technology, Ministry of Education; Department of Microbiology, Nankai University, Tianjin, 300-071, China

    Ming Li, Xiuyuan Ou, Laijun Xing & Mingchun Li

  2. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300-457, China

    Ming Li

  3. Biotechnology Research Center, Jilin Academy of Agriculture Sciences, Changchun, 130-033, China

    Xiangdong Yang, Dongquan Guo & Xueyan Qian

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  1. Ming Li
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  2. Xiuyuan Ou
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  3. Xiangdong Yang
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  4. Dongquan Guo
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  5. Xueyan Qian
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Correspondence to Mingchun Li.

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Li, M., Ou, X., Yang, X. et al. Cloning and identification of a novel C18-Δ9 polyunsaturated fatty acid specific elongase gene from DHA-producing Isochrysis galbana H29. Biotechnol Bioproc E 17, 22–32 (2012). https://doi.org/10.1007/s12257-011-0037-3

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  • DOI: https://doi.org/10.1007/s12257-011-0037-3

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