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Isolation and Functional Characterisation of a fads2 in Rainbow Trout (Oncorhynchus mykiss) with Δ5 Desaturase Activity.
PLoS One 2016; 11(3):e0150770Plos

Abstract

Rainbow trout, Oncorhynchus mykiss, are intensively cultured globally. Understanding their requirement for long-chain polyunsaturated fatty acids (LC-PUFA) and the biochemistry of the enzymes and biosynthetic pathways required for fatty acid synthesis is important and highly relevant in current aquaculture. Most gnathostome vertebrates have two fatty acid desaturase (fads) genes with known functions in LC-PUFA biosynthesis and termed fads1 and fads2. However, teleost fish have exclusively fads2 genes. In rainbow trout, a fads2 cDNA had been previously cloned and found to encode an enzyme with Δ6 desaturase activity. In the present study, a second fads2 cDNA was cloned from the liver of rainbow trout and termed fads2b. The full-length mRNA contained 1578 nucleotides with an open reading frame of 1365 nucleotides that encoded a 454 amino acid protein with a predicted molecular weight of 52.48 kDa. The predicted Fads2b protein had the characteristic traits of the microsomal Fads family, including an N-terminal cytochrome b5 domain containing the heme-binding motif (HPPG), histidine boxes (HDXGH, HFQHH and QIEHH) and three transmembrane regions. The fads2b was expressed predominantly in the brain, liver, intestine and pyloric caeca. Expression of the fasd2b in yeast generated a protein that was found to specifically convert eicosatetraenoic acid (20:4n-3) to eicosapentaenoic acid (20:5n-3), and therefore functioned as a Δ5 desaturase. Therefore, rainbow trout have two fads2 genes that encode proteins with Δ5 and Δ6 desaturase activities, respectively, which enable this species to perform all the desaturation steps required for the biosynthesis of LC-PUFA from C18 precursors.

Authors+Show Affiliations

Deakin University, School of Life and Environmental Sciences, Waurn Ponds, Geelong, Victoria, Australia. Universiti Sains Malaysia, School of Biological Sciences, Penang, Malaysia.University of Stirling, Institute of Aquaculture, School of Natural Sciences, Stirling, Scotland, United Kingdom.University of Stirling, Institute of Aquaculture, School of Natural Sciences, Stirling, Scotland, United Kingdom.University of Stirling, Institute of Aquaculture, School of Natural Sciences, Stirling, Scotland, United Kingdom.Deakin University, School of Life and Environmental Sciences, Waurn Ponds, Geelong, Victoria, Australia.Deakin University, School of Life and Environmental Sciences, Waurn Ponds, Geelong, Victoria, Australia.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26943160

Citation

Abdul Hamid, Noor Khalidah, et al. "Isolation and Functional Characterisation of a Fads2 in Rainbow Trout (Oncorhynchus Mykiss) With Δ5 Desaturase Activity." PloS One, vol. 11, no. 3, 2016, pp. e0150770.
Abdul Hamid NK, Carmona-Antoñanzas G, Monroig Ó, et al. Isolation and Functional Characterisation of a fads2 in Rainbow Trout (Oncorhynchus mykiss) with Δ5 Desaturase Activity. PLoS ONE. 2016;11(3):e0150770.
Abdul Hamid, N. K., Carmona-Antoñanzas, G., Monroig, Ó., Tocher, D. R., Turchini, G. M., & Donald, J. A. (2016). Isolation and Functional Characterisation of a fads2 in Rainbow Trout (Oncorhynchus mykiss) with Δ5 Desaturase Activity. PloS One, 11(3), pp. e0150770. doi:10.1371/journal.pone.0150770.
Abdul Hamid NK, et al. Isolation and Functional Characterisation of a Fads2 in Rainbow Trout (Oncorhynchus Mykiss) With Δ5 Desaturase Activity. PLoS ONE. 2016;11(3):e0150770. PubMed PMID: 26943160.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation and Functional Characterisation of a fads2 in Rainbow Trout (Oncorhynchus mykiss) with Δ5 Desaturase Activity. AU - Abdul Hamid,Noor Khalidah, AU - Carmona-Antoñanzas,Greta, AU - Monroig,Óscar, AU - Tocher,Douglas R, AU - Turchini,Giovanni M, AU - Donald,John A, Y1 - 2016/03/04/ PY - 2015/11/05/received PY - 2016/02/17/accepted PY - 2016/3/5/entrez PY - 2016/3/5/pubmed PY - 2016/7/28/medline SP - e0150770 EP - e0150770 JF - PloS one JO - PLoS ONE VL - 11 IS - 3 N2 - Rainbow trout, Oncorhynchus mykiss, are intensively cultured globally. Understanding their requirement for long-chain polyunsaturated fatty acids (LC-PUFA) and the biochemistry of the enzymes and biosynthetic pathways required for fatty acid synthesis is important and highly relevant in current aquaculture. Most gnathostome vertebrates have two fatty acid desaturase (fads) genes with known functions in LC-PUFA biosynthesis and termed fads1 and fads2. However, teleost fish have exclusively fads2 genes. In rainbow trout, a fads2 cDNA had been previously cloned and found to encode an enzyme with Δ6 desaturase activity. In the present study, a second fads2 cDNA was cloned from the liver of rainbow trout and termed fads2b. The full-length mRNA contained 1578 nucleotides with an open reading frame of 1365 nucleotides that encoded a 454 amino acid protein with a predicted molecular weight of 52.48 kDa. The predicted Fads2b protein had the characteristic traits of the microsomal Fads family, including an N-terminal cytochrome b5 domain containing the heme-binding motif (HPPG), histidine boxes (HDXGH, HFQHH and QIEHH) and three transmembrane regions. The fads2b was expressed predominantly in the brain, liver, intestine and pyloric caeca. Expression of the fasd2b in yeast generated a protein that was found to specifically convert eicosatetraenoic acid (20:4n-3) to eicosapentaenoic acid (20:5n-3), and therefore functioned as a Δ5 desaturase. Therefore, rainbow trout have two fads2 genes that encode proteins with Δ5 and Δ6 desaturase activities, respectively, which enable this species to perform all the desaturation steps required for the biosynthesis of LC-PUFA from C18 precursors. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/26943160/Isolation_and_Functional_Characterisation_of_a_fads2_in_Rainbow_Trout__Oncorhynchus_mykiss__with_Δ5_Desaturase_Activity_ L2 - http://dx.plos.org/10.1371/journal.pone.0150770 DB - PRIME DP - Unbound Medicine ER -