Expression of the eicosapentaenoic acid synthesis gene cluster from Shewanella sp. in a transgenic marine cyanobacterium, Synechococcus sp.
The eicosapentaenoic acid (EPA) synthesis gene cluster isolated from a marine bacterium, Shewanella putrefaciens strain SCRC-2738, was cloned and expressed in the marine cyanobacterium Synechococcus sp. A broad-host-range cosmid vector, pJRD215 (10.2 kb, Smr Kmr), was used to clone a 38 kb insert, pEPA, containing the EPA synthesis gene cluster, creating plasmid pJRDEPA (approx. 48 kb). This plasmid was transferred to the cyanobacterial host at a frequency of 2.2 x 10(-7). Cyanobacterial transconjugants grown at 29 degrees C produced 0.12 mg EPA (g dry weight)-1, whereas those grown at 23 degrees C produced 0.56 mg EPA (g dry weight)-1. The yield was further improved to 0.64 mg (g dry weight)-1 by incubation for 1 d at 17 degrees C. This is believed to be the first successful cloning and expression of such a large heterologous gene cluster in a marine cyanobacterium.
Takeyama
Haruko
H
Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16 Naka-cho, Koganei, Tokyo 184, Japan.
Takeda
Daisuke
D
Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16 Naka-cho, Koganei, Tokyo 184, Japan.
Yazawa
Kazunaga
K
Sagami Chemical Research Center, 4-4-1 Nishi-Ohnuma, Sagamihara, Kanagawa 229, Japan.
Yamada
Akiko
A
Sagami Chemical Research Center, 4-4-1 Nishi-Ohnuma, Sagamihara, Kanagawa 229, Japan.
Matsunaga
Tadashi
T
Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16 Naka-cho, Koganei, Tokyo 184, Japan.
D003227:Conjugation, Genetic; D000458:Cyanobacteria / Q000235:genetics / Q000378:metabolism*; D004041:Dietary Fats; D015118:Eicosapentaenoic Acid / Q000096:biosynthesis* / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D015870:Gene Expression; D005798:Genes, Bacterial; D005818:Genetic Engineering; D006092:Gram-Negative Facultatively Anaerobic Rods / Q000235:genetics*; D008386:Marine Biology; D008969:Molecular Sequence Data; D005810:Multigene Family
D004041:Dietary Fats; D015118:Eicosapentaenoic Acid