title: Identification of a Δ12 fatty acid desaturase from oil palm (Elaeis guineensis Jacq.) involved in the biosynthesis of linoleic acid by heterologous expression in Saccharomyces cerevisiae.
abstract: Oil palm (Elaeis guineensis Jacq.) is one of the highest oil-yield crops in the world. A Δ12-desaturases associated with the primary steps of long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis were successfully cloned from oil palm and their functions identified. The open reading frames (ORFs) of egFAD2 (GenBank accession: KT023602) consisted of 1176bp and code for 391 amino acids. Their deduced polypeptides showed 75-93% identity to microsomal Δ12-desaturases from other higher plants, and each contained the three histidine clusters typical of the catalytic domains of such enzymes. RT-PCR experiment indicated that the egFAD2 gene exhibited the highest accumulation in the mesocarp of fruits at 120-140 DAP (i.e. the fourth period of fruit development) and, despite having different expression levels, the other four stages were at significantly lower levels compared with the fourth stage. Plasmid pYES2-egFAD2 was transformed into Saccharomyces cerevisiae strain INVSc1 using lithium acetate method for expression under the induction of galactose. Yeast cells transformed with plasmid constructs containing egFAD12 produced an appreciable amount of linoleic acids (18:2(Δ9,)(12)), not normally present in wild-type yeast cells, indicating that the genes encoded functional Δ12-desaturase enzymes.
authors.affiliation: Department of Biotechnology, Hainan University, Haikou, Hainan 570228, China.
authors.forename: Ruhao
authors.initials: R
authors.lastname: Sun
authors.affiliation: Department of Biotechnology, Hainan University, Haikou, Hainan 570228, China.
authors.forename: Lingchao
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Biotechnology, Hainan University, Haikou, Hainan 570228, China.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Biotechnology, Hainan University, Haikou, Hainan 570228, China.
authors.forename: Yusheng
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Department of Biotechnology, Hainan University, Haikou, Hainan 570228, China. Electronic address: liddfym@hotmail.com.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Biotechnology, Hainan University, Haikou, Hainan 570228, China. Electronic address: xg-wang@163.com.
authors.forename: Xinguang
authors.initials: X
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D028023:Arecaceae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D019295:Computational Biology; D044943:Fatty Acid Desaturases / Q000302:isolation & purification / Q000378:metabolism*; D005638:Fruit / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D019787:Linoleic Acid / Q000096:biosynthesis*; D008861:Microsomes / Q000378:metabolism; D000073878:Palm Oil; D010802:Phylogeny; D010938:Plant Oils / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D016415:Sequence Alignment; D014170:Transformation, Genetic
chemical_list: D010938:Plant Oils; D000073878:Palm Oil; D019787:Linoleic Acid; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase
keywords: Biosynthesis of linoleic acid; Function analysis; Oil palm (Elaeis guineensis Jacq.); Saccharomyces cerevisiae; Δ12 fatty acid desaturase