Cloning of omega 3 desaturase from cyanobacteria and its use in altering the degree of membrane-lipid unsaturation.
Cyanobacteria respond to a decrease in temperature by desaturating fatty acids of membrane lipids to compensate for the decrease in membrane fluidity. Among various desaturation reactions in cyanobacteria, the desaturation of the omega 3 position of fatty acids is the most sensitive to the change in temperature. In the present study, we isolated a gene, designated desB, for the omega 3 desaturase from the cyanobacterium, Synechocystis sp. PCC 6803. The desB gene encodes a protein a 359 amino-acid residues with molecular mass of 41.9 kDa. The desB gene is transcribed as a monocistronic operon that produced a single transcript of 1.4 kb. The level of the desB transcript in cells grown at 22 degrees C was 10 times higher than that in cells grown at 34 degrees C. In order to manipulate the fatty-acid unsaturation of membrane lipids, the desB gene in Synechocystis sp. PCC 6803 was mutated by insertion of a kanamycin-resistance gene cartridge. The resultant mutant was unable to desaturate fatty acids at the omega 3 position. The desA gene, which encodes the delta 12 desaturase of Synechocystis sp. PCC 6803, and the desB gene were introduced into Synechococcus sp. PCC 7942. Whilst the parent cyanobacterium can only desaturate membrane lipids at the delta 9 position of fatty acids, the resultant transformant was able to desaturate fatty acids of membrane lipids at the delta 9, delta 12 and omega 3 positions. These results confirm the function of the desB gene and demonstrate that it is possible to genetically manipulate the fatty-acid unsaturation of membrane lipids in cyanobacteria.
Sakamoto
T
T
Department of Molecular Biomechanics, Graduate University of Advanced Studies, Okazaki, Japan.
Los
D A
DA
Higashi
S
S
Wada
H
H
Nishida
I
I
Ohmori
M
M
Murata
N
N
D000595:Amino Acid Sequence; D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology / Q000235:genetics*; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005798:Genes, Bacterial / Q000235:genetics; D015698:Genomic Library; D008563:Membrane Lipids / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013696:Temperature
D005227:Fatty Acids; D008563:Membrane Lipids; D012329:RNA, Bacterial; D012333:RNA, Messenger; D044943:Fatty Acid Desaturases; C078565:omega-3 fatty acid desaturase