Isolation and Functional Characterization of a Delta 6-Desaturase Gene from the Pike Eel (Muraenesox cinereus)

Sun Hee Kim; Kyung Hee Roh; Jung-Bong Kim; Kwang-Soo Kim; Nam Shin Kim; Hyun Uk Kim; Kyeong-Ryeol Lee; Jong-Sug Park; Jong-Bum Kim

doi:10.1007/s12275-013-3144-3

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Isolation and Functional Characterization of a Delta 6-Desaturase Gene from the Pike Eel (Muraenesox cinereus): Sun Hee Kim ¹, Kyung Hee Roh ¹, Jung-Bong Kim ¹, Kwang-Soo Kim ², Nam Shin Kim ³, Hyun Uk Kim ¹, Kyeong-Ryeol Lee ¹, Jong-Sug Park ⁴, Jong-Bum Kim ¹; Journal of Microbiology 2013;51(6):807-813.
DOI: https://doi.org/10.1007/s12275-013-3144-3
Published online: October 5, 2013

¹National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Republic of Korea, ²National Institute of Crop Science, Rural Development Administration, Suwon 441-707, Republic of Korea, ³Korean Bioinformation Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea, ⁴R&D Coordination Division, Rural Development Administration, Suwon 441-707, Republic of Korea¹National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Republic of Korea, ²National Institute of Crop Science, Rural Development Administration, Suwon 441-707, Republic of Korea, ³Korean Bioinformation Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea, ⁴R&D Coordination Division, Rural Development Administration, Suwon 441-707, Republic of Korea

Corresponding author: Jong-Bum Kim , Tel: +82-31-299-1702,

Received: 11 March 2013 • Accepted: 12 July 2013

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Abstract

Stearidonic acid (STA; 18:4n-3) and γ-linolenic acid (GLA; 18:3n-6) are significant intermediates in the biosynthetic pathway for the very-long-chain polyunsaturated fatty acids of eicosapentaenoic acid (EPA; 20:5n-3) and arachidonic acid (ARA; 20:4n-6), respectively. To develop a sustainable system for the production of dietary polyunsaturated fatty acids, we focused on the action of the enzyme delta 6-desaturase (D6DES) on the essential acids, linoleic acid (LA; 18:2n-6) and α-linolenic acid (ALA; 18:3n-3). A 1,335-bp full-length cDNA encoding D6DES (McD6DES) was cloned from Muraenesox cinereus using degenerate PCR and RACE-PCR
methods
. To investigate the enzymatic activity of McD6DES in the production of n-6 and n-3 fatty acids, a recombinant plasmid expressing McD6DES (pYES-McD6DES) was transformed into and expressed in Saccharomyces cerevisiae. The exogenously expressed McD6DES produced GLA and STA at conversion rates of 14.2% and 45.9%, respectively, from the exogenous LA and ALA substrates. These results indicate that McD6DES is essentially a delta 6-desaturase involved in very-long-chain polyunsaturated fatty acid synthesis.

Keywords: delta 6-desaturase;γ-linolenic acid;Muraenesox cinereus;Saccharomyces cerevisiae;stearidonic acid

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Isolation and Functional Characterization of a Delta 6-Desaturase Gene from the Pike Eel (Muraenesox cinereus)

J. Microbiol. 2013;51(6):807-813. Published online October 5, 2013

DOI: https://doi.org/10.1007/s12275-013-3144-3

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