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A novel fungal ω3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

A filamentous fungus, Mortierella alpina 1S-4, is capable of producing not only arachidonic acid (AA; 20:4n-6) but also eicosapentaenoic acid (EPA; 20:5n-3) below a cultural temperature of 20°C. Here, we describe the isolation and characterization of a gene (maw3) that encodes a novel ω3-desaturase from M. alpina 1S-4. Based on the conserved sequence information for M. alpina 1S-4 Δ12-desaturase and Saccharomyces kluyveri ω3-desaturase, the ω3-desaturase gene from M. alpina 1S-4 was cloned. Homology analysis of protein databases revealed that the amino acid sequence showed 51% identity, at the highest, with M. alpina 1S-4 Δ12-desaturase, whereas it exhibited 36% identity with Sac. kluyveri ω3-desaturase. The cloned cDNA was confirmed to encode the ω3-desaturase by its expression in the yeast Sac. cerevisiae. Analysis of the fatty acid composition of the yeast transformant demonstrated that 18-carbon and 20-carbon n-3 polyunsaturated fatty acids (PUFAs) were accumulated through conversion of exogenous 18-carbon and 20-carbon n-6 PUFAs. The substrate specificity of the M. alpina 1S-4 ω3-desaturase differs from those of the known fungal ω3-desaturases from Sac. kluyveri and Saprolegnia diclina. Plant, cyanobacterial and Sac. kluyveri ω3-desaturases desaturate 18-carbon n-6 PUFAs, Spr. diclina ω3-desaturase desaturates 20-carbon n-6 PUFAs and Caenorhabditis elegans ω3-desaturase prefers 18-carbon n-6 PUFAs as substrates rather than 20-carbon n-6 PUFAs. The substrate specificity of M. alpina 1S-4 ω3-desaturase is rather similar to that of C. elegans ω3-desaturase, but the M. alpina ω3-desaturase can more effectively convert AA into EPA when expressed in yeast. The M. alpina 1S-4 ω3-desaturase is the first known fungal desaturase that uses both 18-carbon and 20-carbon n-6 PUFAs as substrates.

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Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research (No. 15658024, to S.S.) from the Ministry of Education, Science, Sports and Culture, Japan, by the COE for Microbial-Process Development Pioneering Future Production Systems (COE program of the Ministry of Education, Science, Sports and Culture, Japan, to S.S.) and by the Project of the New Energy and Industrial Technology Development Organization, Japan.

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  1. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto, 606-8502, Japan

    Eiji Sakuradani, Takahiro Abe, Keita Iguchi & Sakayu Shimizu

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  1. Eiji Sakuradani
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  2. Takahiro Abe
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  3. Keita Iguchi
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  4. Sakayu Shimizu
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Correspondence to Sakayu Shimizu.

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Sakuradani, E., Abe, T., Iguchi, K. et al. A novel fungal ω3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4. Appl Microbiol Biotechnol 66, 648–654 (2005). https://doi.org/10.1007/s00253-004-1760-x

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  • DOI: https://doi.org/10.1007/s00253-004-1760-x

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