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Identification of a fatty acid ∆6-desaturase gene from the eicosapentaenoic acid-producing fungus Pythium splendens RBB-5

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Abstract

A new full-length cDNA (PsD6) putatively encoding a ∆6-desaturase was cloned from the eicosapentaenoic acid-producing fungus Pythium splendens RBB-5. PsD6 contained an open reading frame of 1380 bp encoding a protein of 459 amino acids. The deduced amino acid sequence showed high similarity to those of other ∆6-desaturases. A recombinant vector expressing PsD6 (pPIC3.5K-D6) was constructed and transformed into Pichia pastoris GS115. The heterologous expressed PsD6 in P. pastoris desaturated linoleic acid to γ-linolenic acid but not desaturated α-linolenic acid to stearidonic acid. The results indicated that PsD6 was a fatty acid ∆6-desaturase and it had a substrate specificity for linoleic acid.

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References

  • Cho HP, Nakamura MT, Clarke SD (1999) Cloning expression, and nutritional regulation of the mammalian ∆6-desaturase. J Biol Chem 274:471–477

    Article  PubMed  CAS  Google Scholar 

  • Domergur F, Lerchl J, Zähringer U et al (2002) Cloning and functional characterization of Phaeodactylum tricorntum front-end desaturases involved in eicosapentaenoic acid biosynthesis. Eur J Biochem 269:4105–4113

    Article  Google Scholar 

  • Hu J, Zhou P, Zhu Y et al (2011) Isolation of eicosapentaenoic acid-producing fungi from soil based on polymerase chain reaction amplification. Z Naturforsch 66c:429–433

    Google Scholar 

  • Huang YS, Chaudhary S, Thurmond JM et al (1999) Cloning of ∆12- and ∆6-desaturase from Mortierella alpina and recombinant production of γ-linolenic acid in Saccharomyces cerevisiae. Lipids 34:649–658

    Article  PubMed  CAS  Google Scholar 

  • Li YT, Li MT, Fu CH et al (2009) Improvement of arachidonic acid and eicosapentaenoic acid production by increasing the copy number of the genes encoding fatty acid desaturase and elongase into Pichia pastoris. Biotechnol Lett 31:1011–1017

    Article  PubMed  CAS  Google Scholar 

  • Liu J, Li D, Yin Y et al (2011) ∆6-Desaturase from Mortierella alpina: cDNA cloning, expression, and phylogenetic analysis. Biotechnol Lett 33:1985–1991

    Article  PubMed  CAS  Google Scholar 

  • Petrie JR, Shrestha P, Belide S et al (2012) Transgenic production of arachidonic acid in oilseeds. Transgenic Res 21:139–147

    Article  PubMed  CAS  Google Scholar 

  • Qiu X, Hong HP, Datla N et al (2002) Expression of borage ∆6 desaturase in Saccharomyces cerevisiae and oilseed crops. Can J Bot 80:42–49

    Article  CAS  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    PubMed  CAS  Google Scholar 

  • Sayanova OV, Beaudoin F, Michaelson LV et al (2003) Identification of Primula fatty acid ∆6-desaturase with n-3 substrate preferences. FEBS Lett 542:100–104

    Article  PubMed  CAS  Google Scholar 

  • Sayanova O, Haslam R, Qi B et al (2006) The alternate pathway C208-desaturase from the non-photosynthetic organism Acanthamoeba castellanii is an atypical cytochrome b5-fusion desaturase. FEBS Lett 580:1946–1952

    Article  PubMed  CAS  Google Scholar 

  • Shanklin J, White E, Fox BG (1994) Eight histidine residues are catalytically essential in a membrane-associated iron enzyme, stearoyl-CoA desaturase, and are conserved in alkane hydroxylase and xylene monooxygenase. Biochemistry 33:12787–12794

    Article  PubMed  CAS  Google Scholar 

  • Simopoulos AP (1991) Omega-3 fatty acids in health and disease and in growth and development. Am J Clin Nutr 54:438–463

    PubMed  CAS  Google Scholar 

  • Smith MA, Jonsson L, Stymne S et al (1992) Evidence for cytochrome b 5 as an electron donor in ricinoleic acid biosynthesis in microsomal preparations from developing castor bean (Ricinus communis L.). Biochem J 287:141–144

    PubMed  CAS  Google Scholar 

  • Sydenham E, Dangour AD, Lim WS (2012) Omega 3 fatty acid for the prevention of cognitive decline and dementia. Cochrane Data Syst Rev Issue 6 Art No CD005379. doi:10.1002/14651858.CD005379.pub3

  • Tamura K, Dudley J, Nei M et al (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  • Tavares S, Grotkjær T, Obsen T et al (2011) Metabolic engineering of Saccharomyces cerevisiae for production of eicosapentaenoic acid, using a novel ∆5-desaturase from Paramecium tetraurelia. Appl Environ Microbiol 77:1854–1861

    Article  PubMed  CAS  Google Scholar 

  • Tur JA, Bibiloni MM, Sureda A, Pons A (2012) Dietary sources of omega 3 fatty acids: public health risks and benefits. Br J Nutr 107(Suppl S2):S23–S52. doi:10.1017/S0007114512001456

  • Wallis JG, Browse J (1999) The ∆8-desaturase of Euglena gracilis: an alternate pathway for synthesis of 20-carbon polyunsaturated fatty acids. Arch Biochem Biophys 365:307–316

    Article  PubMed  CAS  Google Scholar 

  • Wen ZY, Chen F (2001) Application of statistically-based experimental designs for the optimization of eicosapentaenoic acid production by the diatom Nitzschia laevis. Biotechnol Bioeng 75:159–169

    Article  PubMed  CAS  Google Scholar 

  • Zhu M, Zhou PP, Yu LJ (2002) Extraction of lipids from Mortierella alpina and enrichment of arachidonic acid from the fungal lipids. Bioresour Technol 84:93–95

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 20976065).

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Correspondence to Longjiang Yu.

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Ruijiao Zhang and Yuanmin Zhu contributed equally to this work.

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Zhang, R., Zhu, Y., Ren, L. et al. Identification of a fatty acid ∆6-desaturase gene from the eicosapentaenoic acid-producing fungus Pythium splendens RBB-5. Biotechnol Lett 35, 431–438 (2013). https://doi.org/10.1007/s10529-012-1101-6

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  • DOI: https://doi.org/10.1007/s10529-012-1101-6

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