Expression of Thraustochytrium sp.FJN-10 ?~4-Desaturase Gene in Saccharomyces cerevisiae / 微生物学通报

ABSTRACT

1.6 kb ?4-desaturase gene(FAD4)was amplified by PCR using plasmid pGEM-TFAD4 as template.The fragment was subcloned into the HindⅢ/XbaⅠrestriction site of pYES2.0 vector.Recombinant plasmid pYFAD4 was transformed into Saccharomyces cerevisiae strain INVScl for expression.It was found to exhibit ?4-fatty acid desaturase activity in the recombinant S.cerevisiae YFAD4 in the presence of exogenous fatty acid substrate docosapentaenoic acid(100?mol/L)under introduction of GAL1.Expression of the FAD4 under appropriate media and temperature conditions led to the production of DHA and it reached 41.13% of the total yeast fatty acid by GC detection.It was suggested that the protein encoded by FAD4 could specifically catalyze DPA into DHA.