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Cloning of a $\Delta5$ desaturase from Thraustochytrium sp. 26185 and Functional Expression in Pichia Pastoris  

Chung Tae-Ho (Department of Biological and Chemical Engineering, College of Engineering, Inha University)
Lee Su-Jin (Department of Biological and Chemical Engineering, College of Engineering, Inha University)
Oh Hyo-Jeong (Department of Biological and Chemical Engineering, College of Engineering, Inha University)
Kim Geun-Joong (Department of Biological and Chemical Engineering, College of Engineering, Inha University)
Hur Byung-Ki (Department of Biological and Chemical Engineering, College of Engineering, Inha University)
Publication Information
KSBB Journal / v.20, no.2, 2005 , pp. 93-99 More about this Journal
Abstract
Polyunsaturated fatty acids, that is PUFAs, are important constituents of membranes particularly found in the retina and central nervous system. In microorganism-based PUFAs biosynthesis, the genus Thraustochytrids is well evaluated for their potential as a promising candidate in the practical production of PUFAs, such as AA and DHA. In this study, we attempted to optimize a method of total nucleic acid extraction from this microorganism as a preliminary experiment. Using the extracted nucleic acid and degenerated primers for direct PCR, we isolated a $\Delta5$ desaturase gene that contained 1320-nucleotide and encoded 439 amino acids. This gene exhibited an expected function, when expressed in P. pastoris in the presence of appropriate exogenous substrate, as an evidence for $\Delta5$ desaturase activity (conversion of DGLA to AA). These results and information could provide a basis for the construction of engineered strains suitable for the practical production of PUFAs.
Keywords
Thraustochytrium; polyunsaturated fatty acid; P. pastoris; nucleic acid extraction;
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