Improvement of arachidonic acid and eicosapentaenoic acid production by increasing the copy number of the genes encoding fatty acid desaturase and elongase into Pichia pastoris.
Genes encoding Delta6 desaturase, Delta6 fatty acid elongase, and Delta5 desaturase from the alga, Phaeodactylum tricornutum, were co-expressed in Pichia pastoris to produce arachidonic acid (ARA; 20:4 Delta(5, 8, 11, 14)) and eicosapentaenoic acid (EPA; 20:5 Delta(5, 8, 11, 14, 17)). A panel of Pichia clones carrying progressively increasing copies of the heterologous gene expression cassette was created using an in vitro multimerization approach. ARA and EPA accumulated up to 0.3 and 0.1% of total fatty acids, respectively, in the recombinant P. pastoris carrying with double copies of these three heterologous genes, as compared to 0.1 and 0.05%, respectively, in the recombinant P. pastoris with single copy.
Li
Yun-Tao
YT
Institute of Resource Biology and Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China.
Li
Mao-Teng
MT
Fu
Chu-Hua
CH
Zhou
Peng-Peng
PP
Liu
Jian-Min
JM
Yu
Long-Jiang
LJ
D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D016718:Arachidonic Acid / Q000096:biosynthesis*; D017377:Diatoms / Q000201:enzymology / Q000235:genetics; D015118:Eicosapentaenoic Acid / Q000096:biosynthesis*; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism; D000081031:Fatty Acid Elongases; D018628:Gene Dosage; D005800:Genes, Fungal; D010843:Pichia / Q000201:enzymology* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D016718:Arachidonic Acid; D015118:Eicosapentaenoic Acid; D044943:Fatty Acid Desaturases; D000123:Acetyltransferases; D000081031:Fatty Acid Elongases