title: Coexpression of multiple genes reconstitutes two pathways of very long-chain polyunsaturated fatty acid biosynthesis in Pichia pastoris.
abstract: The introduction of novel traits to cells often requires the stable coexpression of multiple genes within the same cell. Herein, we report that C22 very long-chain polyunsaturated fatty acids (VLC-PUFAs) were synthesized from C18 precursors by reactions catalyzed by delta 6-desaturase, an ELOVL5 involved in VLC-PUFA elongation, and delta 5-desaturase. The coexpression of McD6DES, AsELOVL5, and PtD5DES encoding the corresponding enzymes, produced docosatetraenoic acid (C22:4 n-6) and docosapentaenoic acid (C22:5 n-3), as well as arachidonic acid (C20:4 n-6) and eicosapentaenoic acid (C20:5 n-3) in the methylotrophic yeast Pichia pastoris. The expression of each gene increased within 24 h, with high transcript levels after induction with 0.5 or 1 % methanol. High levels of the newly expressed VLC-PUFAs occurred after 144 h. This expression system exemplifies the recent progress and future possibilities of the metabolic engineering of VLC-PUFAs in oilseed crops.
authors.affiliation: National Academy of Agricultural Science, Rural Development Administration, Seodun-Dong, Suwon, 441-707, Korea, sunheekim00@korea.kr.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Kim
authors.forename: Kyung Hee
authors.initials: KH
authors.lastname: Roh
authors.forename: Kwang-Soo
authors.initials: KS
authors.lastname: Kim
authors.forename: Hyun Uk
authors.initials: HU
authors.lastname: Kim
authors.forename: Kyeong-Ryeol
authors.initials: KR
authors.lastname: Lee
authors.forename: Han-Chul
authors.initials: HC
authors.lastname: Kang
authors.forename: Jong-Bum
authors.initials: JB
authors.lastname: Kim
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics*; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D015870:Gene Expression; D060847:Metabolic Engineering; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D013997:Time Factors
chemical_list: D005231:Fatty Acids, Unsaturated