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KMID : 0880220130510060807
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Journal of Microbiology
2013 Volume.51 No. 6 p.807 ~ p.813
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Isolation and Functional Characterization of a Delta 6-Desaturase Gene from the Pike Eel (Muraenesox cinereus)
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Kim Sun-Hee
Roh Kyung-Hee
Kim Nam-Shin
Park Jong-Sug
Kim Jung-Bong
Kim Kwang-Soo
Kim Hyun-Uk
Lee Kyeong-Ryeol
Kim Jong-Bum
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Abstract
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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.
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KEYWORD
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delta 6-desaturase, γ-linolenic acid, Muraenesox cinereus, Saccharomyces cerevisiae, stearidonic acid
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