Isolation and functional characterization of a delta 6-desaturase gene from the pike eel (Muraenesox cinereus)

• Published in: The journal of microbiology 2013, 51(6), , pp.807-813
• Main Authors: Kim, Sun Hee, Roh, Kyung Hee, Kim, Jung-Bong, Kim, Kwang-Soo, Kim, Nam Shin, Kim, Hyun Uk, Lee, Kyeong-Ryeol, Park, Jong-Sug, Kim, Jong-Bum
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Biomedical and Life Sciences; Brackish; Cloning; Commercial fishing; Crops; Eels - genetics; Enzymatic activity; Enzymes; Fatty acids; Fatty Acids, Omega-3 - metabolism; Fish; Fish Proteins - chemistry; Fish Proteins - genetics; Fish Proteins - isolation & purification; Fish Proteins - metabolism; gamma-Linolenic Acid - metabolism; Genomics and Molecular Biology; Life Sciences; Linoleoyl-CoA Desaturase - chemistry; Linoleoyl-CoA Desaturase - genetics; Linoleoyl-CoA Desaturase - isolation & purification; Linoleoyl-CoA Desaturase - metabolism; Microbial Genetics; Microbiology; Molecular Sequence Data; Muraenesox cinereus; Oils & fats; Oilseeds; Polyunsaturated fatty acids; Rural development; Saccharomyces cerevisiae; Sequence Alignment; Yeast; 생물학
• ISSN: 1225-8873; 1976-3794
• DOI: 10.1007/s12275-013-3144-3

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.