Isolation and characterization of delta(6)-desaturase, an ELO-like enzyme and delta(5)-desaturase from the liverwort Marchantia polymorpha and production of arachidonic and eicosapentaenoic acids in the methylotrophic yeast Pichia pastoris.
The liverwort Marchantia polymorpha contains high proportions of arachidonic and eicosapentaenoic acids. In general, these C20 polyunsaturated fatty acids (PUFA) are synthesized from linoleic and alpha -linolenic acids, respectively, by a series of reactions catalyzed by Delta(6)-desaturase, an ELO-like enzyme involved in Delta(6) elongation and Delta(5)-desaturase. Here we report the isolation and characterization of the cDNAs, MpDES6, MpELO1 and MpDES5, coding for the respective enzymes from M. polymorpha. Co-expression of the MpDES6, MpELO1 and MpDES5 cDNAs resulted in the accumulation of arachidonic and eicosapentaenoic acids in the methylotrophic yeast Pichia pastoris. Interestingly, Delta(6) desaturation by the expression of the MpDES6 cDNA appears to occur both in glycerolipids and the acyl-CoA pool, although other lower-plant Delta(6)-desaturases are known to have a strong preference for glycerolipids.
Laboratory of Plant Molecular Biology, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kitashirakawaoiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
Masataka
M
Kajikawa
Katsuyuki T
KT
Yamato
Yoshito
Y
Kohzu
Masutoshi
M
Nojiri
Eiji
E
Sakuradani
Sakayu
S
Shimizu
Yasuyoshi
Y
Sakai
Hideya
H
Fukuzawa
Kanji
K
Ohyama
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000595:Amino Acid Sequence; D016718:Arachidonic Acid / Q000032:analysis / Q000096:biosynthesis*; D000090943:Delta-5 Fatty Acid Desaturase; D015118:Eicosapentaenoic Acid / Q000032:analysis / Q000096:biosynthesis*; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000096:biosynthesis; D015971:Gene Expression Regulation, Enzymologic; D035926:Hepatophyta / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D050602:Linoleoyl-CoA Desaturase; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010802:Phylogeny; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D017962:alpha-Linolenic Acid / Q000096:biosynthesis
D000090943:Delta-5 Fatty Acid Desaturase; D005231:Fatty Acids, Unsaturated; D011994:Recombinant Proteins; D017962:alpha-Linolenic Acid; D016718:Arachidonic Acid; D000105:Acetyl Coenzyme A; D015118:Eicosapentaenoic Acid; D044943:Fatty Acid Desaturases; D050602:Linoleoyl-CoA Desaturase