Cloning, expression and stress-respondent transcription of long-chain acyl-coenzyme A synthetase cDNA gene of Nannochloropsis gaditana and its involvement in the biosynthesis of eicosapentaenoic and decosahexaenoic acids.
A novel cDNA gene, NgLACS, that encodes a long-chain acyl-CoA sythetase (LACS), was cloned from Nannochloropsis gaditana and characterized. The cDNA was 2,360 bp in length, consisting of an ORF of 1,950 bp, a 5'-untranslated region of 88 bp and a 3'-untranslated region of 322 bp. The deduced amino acid sequence of LACS was 649 amino acid residues in length with a predicted molecular weight of 71 kDa and an isoelectric point of pH 7.8. When the alga was treated with excessive nitrogen and iron, and at 15 °C, the proportion of long-chain polyunsaturated acyl-CoAs in the total acyl-CoAs and the abundance of NgLACS cDNA gene transcript were up-regulated. Over-expression of NgLACS in Saccharomyces cerevisiae caused the accumulation of eicosapentaenoic acid and docosahexaenoic acid.
The First Institute of Oceanography, State Oceanic Administration of China, Qingdao, 266061, People's Republic of China, zmg@fio.org.cn.
Ming-Gang
MG
Zheng
Yi-Jiang
YJ
Huang
Li
L
Zheng
Zhong-Tao
ZT
Sun
Ling
L
Wang
D003066:Coenzyme A Ligases / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004281:Docosahexaenoic Acids / Q000378:metabolism*; D015118:Eicosapentaenoic Acid / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D058009:Stramenopiles / Q000201:enzymology* / Q000235:genetics; D013312:Stress, Physiological
D018076:DNA, Complementary; D011994:Recombinant Proteins; D004281:Docosahexaenoic Acids; D015118:Eicosapentaenoic Acid; D003066:Coenzyme A Ligases