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A novel fungal omega3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4.
Appl Microbiol Biotechnol 2005; 66(6):648-54AM

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 degrees C. Here, we describe the isolation and characterization of a gene (maw3) that encodes a novel omega3-desaturase from M. alpina 1S-4. Based on the conserved sequence information for M. alpina 1S-4 Delta12-desaturase and Saccharomyces kluyveri omega3-desaturase, the omega3-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 Delta12-desaturase, whereas it exhibited 36% identity with Sac. kluyveri omega3-desaturase. The cloned cDNA was confirmed to encode the omega3-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 omega3-desaturase differs from those of the known fungal omega3-desaturases from Sac. kluyveri and Saprolegnia diclina. Plant, cyanobacterial and Sac. kluyveri omega3-desaturases desaturate 18-carbon n-6 PUFAs, Spr. diclina omega3-desaturase desaturates 20-carbon n-6 PUFAs and Caenorhabditis elegans omega3-desaturase prefers 18-carbon n-6 PUFAs as substrates rather than 20-carbon n-6 PUFAs. The substrate specificity of M. alpina 1S-4 omega3-desaturase is rather similar to that of C. elegans omega3-desaturase, but the M. alpina omega3-desaturase can more effectively convert AA into EPA when expressed in yeast. The M. alpina 1S-4 omega3-desaturase is the first known fungal desaturase that uses both 18-carbon and 20-carbon n-6 PUFAs as substrates.

Authors+Show Affiliations

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto, 606-8502, Japan.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

15538555

Citation

Sakuradani, Eiji, et al. "A Novel Fungal Omega3-desaturase With Wide Substrate Specificity From Arachidonic Acid-producing Mortierella Alpina 1S-4." Applied Microbiology and Biotechnology, vol. 66, no. 6, 2005, pp. 648-54.
Sakuradani E, Abe T, Iguchi K, et al. A novel fungal omega3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4. Appl Microbiol Biotechnol. 2005;66(6):648-54.
Sakuradani, E., Abe, T., Iguchi, K., & Shimizu, S. (2005). A novel fungal omega3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4. Applied Microbiology and Biotechnology, 66(6), pp. 648-54.
Sakuradani E, et al. A Novel Fungal Omega3-desaturase With Wide Substrate Specificity From Arachidonic Acid-producing Mortierella Alpina 1S-4. Appl Microbiol Biotechnol. 2005;66(6):648-54. PubMed PMID: 15538555.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel fungal omega3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4. AU - Sakuradani,Eiji, AU - Abe,Takahiro, AU - Iguchi,Keita, AU - Shimizu,Sakayu, Y1 - 2004/11/05/ PY - 2004/07/12/received PY - 2004/08/31/accepted PY - 2004/08/23/revised PY - 2004/11/13/pubmed PY - 2005/8/5/medline PY - 2004/11/13/entrez SP - 648 EP - 54 JF - Applied microbiology and biotechnology JO - Appl. Microbiol. Biotechnol. VL - 66 IS - 6 N2 - 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 degrees C. Here, we describe the isolation and characterization of a gene (maw3) that encodes a novel omega3-desaturase from M. alpina 1S-4. Based on the conserved sequence information for M. alpina 1S-4 Delta12-desaturase and Saccharomyces kluyveri omega3-desaturase, the omega3-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 Delta12-desaturase, whereas it exhibited 36% identity with Sac. kluyveri omega3-desaturase. The cloned cDNA was confirmed to encode the omega3-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 omega3-desaturase differs from those of the known fungal omega3-desaturases from Sac. kluyveri and Saprolegnia diclina. Plant, cyanobacterial and Sac. kluyveri omega3-desaturases desaturate 18-carbon n-6 PUFAs, Spr. diclina omega3-desaturase desaturates 20-carbon n-6 PUFAs and Caenorhabditis elegans omega3-desaturase prefers 18-carbon n-6 PUFAs as substrates rather than 20-carbon n-6 PUFAs. The substrate specificity of M. alpina 1S-4 omega3-desaturase is rather similar to that of C. elegans omega3-desaturase, but the M. alpina omega3-desaturase can more effectively convert AA into EPA when expressed in yeast. The M. alpina 1S-4 omega3-desaturase is the first known fungal desaturase that uses both 18-carbon and 20-carbon n-6 PUFAs as substrates. SN - 0175-7598 UR - https://www.unboundmedicine.com/medline/citation/15538555/A_novel_fungal_omega3_desaturase_with_wide_substrate_specificity_from_arachidonic_acid_producing_Mortierella_alpina_1S_4_ L2 - https://dx.doi.org/10.1007/s00253-004-1760-x DB - PRIME DP - Unbound Medicine ER -