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Molecular cloning of a Pinguiochrysis pyriformis oleate-specific microsomal {Delta}12-fatty acid desaturase and functional analysis in yeasts and thraustochytrids.
MedLine Citation:
PMID:  21705343     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
We isolated a putative desaturase gene from a marine alga, Pinguiochrysis pyriformis MBIC 10872, which is capable of accumulating eicosapentaenoic acid (C20:5(Δ5,8,11,14,17)). The gene possessed an open reading frame of 1,314 bp encoding a putative 437 amino acid residues showing high sequence identity (37-48%) with fungal and nematode Δ12-fatty acid desaturases. Yeast cells transformed with the gene converted endogenous oleic acid (C18:1(Δ9)) to linoleic acid (C18:2(Δ9,12)). However, no double bonds were introduced into other endogenous fatty acids or exogenously added fatty acids. Flag-tagged enzyme was recovered in the micosome fraction when expressed in yeast cells. To express the gene in thraustochytrids, a construct driven by the thraustochytrid-derived ubiquitin promoter was used. Interestingly, exogenously added oleic acid was converted to linoleic acid in the gene transformants but not mock transformants of Aurantiochytrium limacinum mh0186. These results clearly indicate that the gene encodes a microsomal Δ12-fatty acid desaturase and was expressed functionally in not only yeasts but also thraustochytrids. This is the first report describing the heterozygous expression of a fatty acid desaturase in thraustochytrids, and could facilitate a genetic approach toward fatty acid synthesis in thraustochytrids which are expected to be an alternative source of polyunsaturated fatty acids.
Authors:
Takanori Matsuda; Keishi Sakaguchi; Takumi Kobayashi; Eriko Abe; Norihide Kurano; Akira Sato; Yuji Okita; Shinichi Sugimoto; Yoichiro Hama; Masahiro Hayashi; Nozomu Okino; Makoto Ito
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-6-23
Journal Detail:
Title:  Journal of biochemistry     Volume:  -     ISSN:  1756-2651     ISO Abbreviation:  -     Publication Date:  2011 Jun 
Date Detail:
Created Date:  2011-6-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0376600     Medline TA:  J Biochem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan; Marine Biotechnology Institute Co., Ltd. 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan; Nippon Suisan Kaisha, Ltd., 6-2 Otemachi 2-chome, Chiyoda-ku, Tokyo 100-8686, Japan; Faculty of Agriculture, Saga University, 1 Honjo, Saga 840-8502, Japan; Faculty of Agriculture, Miyazaki University, Miyazaki 889-2192, Japan; Bio-Architecture Center, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
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