Isolation and characterization of Δ6-desaturase, an ELO-like enzyme and Δ5-desaturase from the liverwort Marchantia polymorpha and production of arachidonic and eicosapentaenoic acids in the methylotrophic yeast Pichia pastoris

被引:49
作者
Kajikawa, M
Yamato, KT
Kohzu, Y
Nojiri, M
Sakuradani, E
Shimizu, S
Sakai, Y
Fukuzawa, H
Ohyama, K
机构
[1] Kyoto Univ, Grad Sch Biostudies, Div Integrated Life Sci, Plant Mol Biol Lab,Sakyo Ku, Kyoto 6068502, Japan
[2] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Lab Fermentat Physiol & Appl Microbiol, Kyoto 6068502, Japan
[3] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Lab Microbial Biotechnol, Kyoto 6068502, Japan
基金
日本学术振兴会;
关键词
acyl-CoA; bryophyte; fatty acid desaturation; fatty acid elongation; heterologous gene expression; polyunsaturated fatty acid;
D O I
10.1023/B:PLAN.0000036366.57794.ee
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. Coexpression of the MpDES6, MpELO1 and MpDES5 cDNAs resulted in the accumulation of arachidonic and eicosapentaenoic acids in the methylotrophic yeast Pichia pastoris. Interestingly, D6 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.
引用
收藏
页码:335 / 352
页数:18
相关论文
共 51 条
[1]   Molecular cloning and functional characterization of rat Δ-6 fatty acid desaturase [J].
Aki, T ;
Shimada, Y ;
Inagaki, K ;
Higashimoto, H ;
Kawamoto, S ;
Shigeta, S ;
Ono, K ;
Suzuki, O .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 255 (03) :575-579
[2]   Heterologous reconstitution in yeast of the polyunsaturated fatty acid biosynthetic pathway [J].
Beaudoin, F ;
Michaelson, LV ;
Hey, SJ ;
Lewis, MJ ;
Shewry, PR ;
Sayanova, O ;
Napier, JA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (12) :6421-6426
[3]   Production of fatty acid components of meadowfoam oil in somatic soybean embryos [J].
Cahoon, EB ;
Marillia, EF ;
Stecca, KL ;
Hall, SE ;
Taylor, DC ;
Kinney, AJ .
PLANT PHYSIOLOGY, 2000, 124 (01) :243-251
[4]   Cloning, expression, and nutritional regulation of the mammalian Δ-6 desaturase [J].
Cho, HP ;
Nakamura, MT ;
Clarke, SD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (01) :471-477
[5]   Cloning, expression, and fatty acid regulation of the human Δ-5 desaturase [J].
Cho, HP ;
Nakamura, M ;
Clarke, SD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (52) :37335-37339
[6]   LIPIDS OF BRYOPHYTES [J].
DEMBITSKY, VM .
PROGRESS IN LIPID RESEARCH, 1993, 32 (03) :281-356
[7]   Acyl carriers used as substrates by the desaturases and elongases involved in very long-chain polyunsaturated fatty acids biosynthesis reconstituted in yeast [J].
Domergue, F ;
Abbadi, A ;
Ott, C ;
Zank, TK ;
Zähringer, U ;
Heinz, E .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (37) :35115-35126
[8]   Cloning and functional characterization of Phaeodactylum tricornutum front-end desaturases involved in eicosapentaenoic acid biosynthesis [J].
Domergue, F ;
Lerchl, J ;
Zähringer, U ;
Heinz, E .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (16) :4105-4113
[9]   Metabolic engineering of fatty acids for breeding of new oilseed crops: strategies, problems and first results [J].
Drexler, H ;
Spiekermann, P ;
Meyer, A ;
Domergue, F ;
Zank, T ;
Sperling, P ;
Abbadi, A ;
Heinz, E .
JOURNAL OF PLANT PHYSIOLOGY, 2003, 160 (07) :779-802
[10]  
Fujisawa M, 2001, GENETICS, V159, P981