Cloning and functional characterization of δ6 fatty acid desaturase (FADS2) in Eurasian perch (Perca fluviatilis)

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)112-125
Number of pages14
JournalComparative Biochemistry and Physiology Part - B: Biochemistry and Molecular Biology
Volume191
DOIs
Publication statusPublished - 1 Jan 2016

Fingerprint

Fatty Acid Desaturases
Perches
Cloning
Unsaturated Fatty Acids
Organism Cloning
Fresh Water
Liver
Intestines
Cytochromes b5
Amino Acids
Aquaculture
Plant Oils
Biosynthesis
Alternative Splicing
Gene expression
Yeast
Open Reading Frames
Real-Time Polymerase Chain Reaction
Amino Acid Sequence
Assays

Keywords

  • Eurasian perch
  • Fatty acid desaturase 2
  • Freshwater fish
  • Highly unsaturated fatty acids
  • Tissue expression

Cite this

@article{22172a419a224d24a6d804026c369f23,
title = "Cloning and functional characterization of δ6 fatty acid desaturase (FADS2) in Eurasian perch (Perca fluviatilis)",
abstract = "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.",
keywords = "Eurasian perch, Fatty acid desaturase 2, Freshwater fish, Highly unsaturated fatty acids, Tissue expression",
author = "F. Geay and E. Tinti and J. Mellery and C. Michaux and Y. Larondelle and E. Perp{\`e}te and P. Kestemont",
year = "2016",
month = "1",
day = "1",
doi = "10.1016/j.cbpb.2015.10.004",
language = "English",
volume = "191",
pages = "112--125",
journal = "Comparative Biochemistry and Physiology Part - B: Biochemistry and Molecular Biology",
issn = "1096-4959",
publisher = "Elsevier Inc.",

}

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

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

SN - 1096-4959

ER -