TY - JOUR
T1 - Cloning and functional characterization of δ6 fatty acid desaturase (FADS2) in Eurasian perch (Perca fluviatilis)
AU - Geay, F.
AU - Tinti, E.
AU - Mellery, J.
AU - Michaux, C.
AU - Larondelle, Y.
AU - Perpète, E.
AU - Kestemont, P.
PY - 2016/1/1
Y1 - 2016/1/1
N2 - The Eurasian perch (Perca fluviatilis) is a freshwater carnivorous species of high interest to diversify inland aquaculture. However, little is known about its ability to bioconvert polyunsaturated fatty acids (PUFAs) from plant oils into long chain polyunsaturated fatty acids (LC-PUFAs). In this study, special attention has been given to the fatty acid desaturase 2 (FADS2) which is commonly described to be a rate-limiting enzyme of the LC-PUFA biosynthesis. This work reports on the cloning, tissue expression and functional characterization of the Eurasian perch fads2, but also on the cloning of two alternative splicing transcripts named fads2-AS1 and fads2-AS2. The fads2 cDNA cloned is composed of an open reading frame (ORF) of 1338 nucleotides (nt) and encodes a protein of 445 amino acids. This deduced amino acid sequence displays the typical structure of microsomal FADS2 including two transmembrane domains and an N-terminal cytochrome b5 domain with the "HPGG" motif. Quantitative real-time PCR assay of fads2, fads2-AS1 and fads2-AS2 expressions revealed that the fads2 transcript was mainly expressed in the liver and intestine and exhibited a typical gene expression pattern of freshwater species while fads2-AS1 and fads2-AS2 genes were highly expressed in the brain, followed by the liver and intestine. Functional characterization of Eurasian perch FADS2 in transgenic yeast showed a fully functional δ6 desaturation activity toward C18 PUFA substrates, without residual δ5 and δ8 desaturase activities.
AB - The Eurasian perch (Perca fluviatilis) is a freshwater carnivorous species of high interest to diversify inland aquaculture. However, little is known about its ability to bioconvert polyunsaturated fatty acids (PUFAs) from plant oils into long chain polyunsaturated fatty acids (LC-PUFAs). In this study, special attention has been given to the fatty acid desaturase 2 (FADS2) which is commonly described to be a rate-limiting enzyme of the LC-PUFA biosynthesis. This work reports on the cloning, tissue expression and functional characterization of the Eurasian perch fads2, but also on the cloning of two alternative splicing transcripts named fads2-AS1 and fads2-AS2. The fads2 cDNA cloned is composed of an open reading frame (ORF) of 1338 nucleotides (nt) and encodes a protein of 445 amino acids. This deduced amino acid sequence displays the typical structure of microsomal FADS2 including two transmembrane domains and an N-terminal cytochrome b5 domain with the "HPGG" motif. Quantitative real-time PCR assay of fads2, fads2-AS1 and fads2-AS2 expressions revealed that the fads2 transcript was mainly expressed in the liver and intestine and exhibited a typical gene expression pattern of freshwater species while fads2-AS1 and fads2-AS2 genes were highly expressed in the brain, followed by the liver and intestine. Functional characterization of Eurasian perch FADS2 in transgenic yeast showed a fully functional δ6 desaturation activity toward C18 PUFA substrates, without residual δ5 and δ8 desaturase activities.
KW - Eurasian perch
KW - Fatty acid desaturase 2
KW - Freshwater fish
KW - Highly unsaturated fatty acids
KW - Tissue expression
UR - http://www.scopus.com/inward/record.url?scp=84945157299&partnerID=8YFLogxK
U2 - 10.1016/j.cbpb.2015.10.004
DO - 10.1016/j.cbpb.2015.10.004
M3 - Article
AN - SCOPUS:84945157299
SN - 1096-4959
VL - 191
SP - 112
EP - 125
JO - Comparative Biochemistry and Physiology. Part B: Biochemistry and Molecular Biology
JF - Comparative Biochemistry and Physiology. Part B: Biochemistry and Molecular Biology
ER -
