Cloning of delta12- and delta6-desaturases from Mortierella alpina and recombinant production of gamma-linolenic acid in Saccharomyces cerevisiae.
Two cDNA clones with homology to known desaturase genes were isolated from the fungus Mortierella alpina. The open reading frame in one clone encoded 399 amino acids and exhibited delta12-desaturase activity when expressed in Saccharomyces cerevisiae in the presence of endogenous fatty acid substrate oleic acid. The insert in another clone contained an open reading frame encoding 457 amino acids and exhibited delta6-desaturase activity in S. cerevisiae in the presence of exogenous fatty acid substrate linoleic acid. Expression of the delta12-desaturase gene under appropriate media and temperature conditions led to the production of linoleic acid at levels up to 25% of the total fatty acids in yeast. When linoleic acid was provided as an exogenous substrate to the yeast cultures expressing the delta6-desaturase activity, the level of gamma-linolenic acid reached 10% of the total yeast fatty acids. Co-expression of both the delta6- and delta12-desaturase cDNA resulted in the endogenous production of gamma-linolenic acid. The yields of gamma-linolenic acid reached as high as 8% of total fatty acids in yeast.
Huang
Y S
YS
Ross Products Division, Abbott Laboratories, Columbus, Ohio 43215, USA.
Chaudhary
S
S
Thurmond
J M
JM
Bobik
E G
EG
Yuan
L
L
Chan
G M
GM
Kirchner
S J
SJ
Mukerji
P
P
Knutzon
D S
DS
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004952:Esters; D044943:Fatty Acid Desaturases / Q000235:genetics*; D005227:Fatty Acids / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D008969:Molecular Sequence Data; D020094:Mortierella / Q000235:genetics*; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D017965:gamma-Linolenic Acid / Q000096:biosynthesis*
D017931:DNA Primers; D018076:DNA, Complementary; D004952:Esters; D005227:Fatty Acids; D017965:gamma-Linolenic Acid; D044943:Fatty Acid Desaturases