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Functional characterization of polyunsaturated fatty acid delta 6-desaturase and elongase genes from the black seabream (Acanthopagrus schlegelii).
Cell Biochem Biophys 2014; 68(2):335-46CB

Abstract

Fatty acid delta 6-desaturase (D6DES) and elongases are key enzymes in the synthesis of polyunsaturated fatty acids (PUFAs) including arachidonic acid (ARA) and eicosapentaenoic acid (EPA) from microorganisms to higher animals. To identify the genes encoding D6DES and elongases for PUFAs, we isolated each cDNA with a high similarity to the D6DES and ELOVL5-like elongases of mammals and fishes via degenerate PCR and RACE-PCR from Acanthopagrus schlegelii. A recombinant vector expressing AsD6DES was subsequently constructed and transformed into Saccharomyces cerevisiae to test the enzymatic activity toward n-6 and n-3 fatty acids in the PUFA biosynthesis. The heterologously expressed AsD6DES produced γ-linolenic acid (GLA, C18:3 n-6) and stearidonic acid (STA, C18:4 n-3) at conversion rates of 26.3-35.6% from exogenous linoleic acid (LA, C18:2 n-6) and α-linolenic acid (ALA, C18:3 n-3) substrates, respectively. When AsELOVL5 was expressed in yeast, it conferred an ability to elongate GLA to di-homo-γ-linolenic acid (DGLA, C20:3 n-6). In addition, AsELOVL5 showed an ability to convert ARA (C20:4 n-6) and EPA (C20:5 n-3) to dodecylthioacetic acid (DTA, C22:4 n-6) and docosapentaenoic acid (DPA, C22:5 n-3), respectively. In these results, the AsD6DES encodes a delta 6-fatty acid desaturase and the AsELOVL5 encoding a long-chain fatty acid elongase shows activity to enlongate C18Δ6/C20Δ5, but not C22.

Authors+Show Affiliations

Rural Development Administration, National Academy of Agricultural Science, Seodun-Dong, Suwon, 441-707, Korea, sunheekim00@korea.kr.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo 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

24052399

Citation

Kim, Sun Hee, et al. "Functional Characterization of Polyunsaturated Fatty Acid Delta 6-desaturase and Elongase Genes From the Black Seabream (Acanthopagrus Schlegelii)." Cell Biochemistry and Biophysics, vol. 68, no. 2, 2014, pp. 335-46.
Kim SH, Park JS, Kim SY, et al. Functional characterization of polyunsaturated fatty acid delta 6-desaturase and elongase genes from the black seabream (Acanthopagrus schlegelii). Cell Biochem Biophys. 2014;68(2):335-46.
Kim, S. H., Park, J. S., Kim, S. Y., Kim, J. B., Roh, K. H., Kim, H. U., ... Kim, J. B. (2014). Functional characterization of polyunsaturated fatty acid delta 6-desaturase and elongase genes from the black seabream (Acanthopagrus schlegelii). Cell Biochemistry and Biophysics, 68(2), pp. 335-46. doi:10.1007/s12013-013-9714-5.
Kim SH, et al. Functional Characterization of Polyunsaturated Fatty Acid Delta 6-desaturase and Elongase Genes From the Black Seabream (Acanthopagrus Schlegelii). Cell Biochem Biophys. 2014;68(2):335-46. PubMed PMID: 24052399.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional characterization of polyunsaturated fatty acid delta 6-desaturase and elongase genes from the black seabream (Acanthopagrus schlegelii). AU - Kim,Sun Hee, AU - Park,Jong Sug, AU - Kim,So Yun, AU - Kim,Jung-Bong, AU - Roh,Kyung Hee, AU - Kim,Hyun Uk, AU - Lee,Kyeong-Ryeol, AU - Kim,Jong Bum, PY - 2013/9/21/entrez PY - 2013/9/21/pubmed PY - 2014/10/10/medline SP - 335 EP - 46 JF - Cell biochemistry and biophysics JO - Cell Biochem. Biophys. VL - 68 IS - 2 N2 - Fatty acid delta 6-desaturase (D6DES) and elongases are key enzymes in the synthesis of polyunsaturated fatty acids (PUFAs) including arachidonic acid (ARA) and eicosapentaenoic acid (EPA) from microorganisms to higher animals. To identify the genes encoding D6DES and elongases for PUFAs, we isolated each cDNA with a high similarity to the D6DES and ELOVL5-like elongases of mammals and fishes via degenerate PCR and RACE-PCR from Acanthopagrus schlegelii. A recombinant vector expressing AsD6DES was subsequently constructed and transformed into Saccharomyces cerevisiae to test the enzymatic activity toward n-6 and n-3 fatty acids in the PUFA biosynthesis. The heterologously expressed AsD6DES produced γ-linolenic acid (GLA, C18:3 n-6) and stearidonic acid (STA, C18:4 n-3) at conversion rates of 26.3-35.6% from exogenous linoleic acid (LA, C18:2 n-6) and α-linolenic acid (ALA, C18:3 n-3) substrates, respectively. When AsELOVL5 was expressed in yeast, it conferred an ability to elongate GLA to di-homo-γ-linolenic acid (DGLA, C20:3 n-6). In addition, AsELOVL5 showed an ability to convert ARA (C20:4 n-6) and EPA (C20:5 n-3) to dodecylthioacetic acid (DTA, C22:4 n-6) and docosapentaenoic acid (DPA, C22:5 n-3), respectively. In these results, the AsD6DES encodes a delta 6-fatty acid desaturase and the AsELOVL5 encoding a long-chain fatty acid elongase shows activity to enlongate C18Δ6/C20Δ5, but not C22. SN - 1559-0283 UR - https://www.unboundmedicine.com/medline/citation/24052399/Functional_characterization_of_polyunsaturated_fatty_acid_delta_6_desaturase_and_elongase_genes_from_the_black_seabream__Acanthopagrus_schlegelii__ L2 - https://dx.doi.org/10.1007/s12013-013-9714-5 DB - PRIME DP - Unbound Medicine ER -