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KMID : 0380020050200020093
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Korean Journal of Biotechnology and Bioengineering
2005 Volume.20 No. 2 p.93 ~ p.99
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Cloning of a Δ5 desaturase from Thraustochytrium sp. 26185 and Functional Expression in Pichia pastoris
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Jung Tae-Ho
Lee Soo-Jin
Oh Hyo-Jeong
Kim Geun-Joong
Hur Byung-Ki
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Abstract
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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 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 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.
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KEYWORD
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Thraustochytrium, polyunsaturated fatty acid, Δ5 desaturase, P. pastoris, nucleic acid extraction
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