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Identification of a novel type of polyunsaturated fatty acid synthase involved in arachidonic acid biosynthesis
Authors:Tetsuro Ujihara  Megumi Nagano  Hajime Wada  Satoshi Mitsuhashi
Institution:1. Kyowa Hakko Bio Co. Ltd., Bioprocess Development Center, Tsukuba, Ibaraki 305-0841, Japan;2. Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan
Abstract:Arachidonic acid (ARA) is a polyunsaturated fatty acid (PUFA) and an essential component of membrane lipids. However, the PUFA synthase required for ARA biosynthesis has not been identified in any organism. To identify the PUFA synthase producing ARA, we determined the draft genome sequence of the marine bacterium Aureispira marina, which produces a high level of ARA, and found a gene cluster encoding a putative PUFA synthase for ARA production. Expression of the gene cluster in Escherichia coli induced production of ARA, demonstrating that the gene cluster encodes a PUFA synthase required for ARA biosynthesis.
Keywords:ACP  acyl-carrier protein  ARA  arachidonic acid  AT  acyltransferase  DHA  docosahexaenoic acid  EPA  eicosapentaenoic acid  ER  enoyl-ACP reductase  GC  gas chromatography  GC&ndash  MS  GC&ndash  mass spectrometry  DH  3-hydroxyacyl-ACP dehydratase  FAME  fatty acid methyl ester  IPTG  isopropyl β-d-1-thiogalactopyranoside  KR  3-ketoacyl-ACP reductase  KS  3-ketoacyl-ACP synthase  LB  Luria&ndash  Bertani  MAT  malonyl-CoA:ACP transacylase  PCR  polymerase chain reaction  ppTase  phosphopantetheinyl transferase  PUFA  polyunsaturated fatty acid  X:Y(z)  fatty acid containing X carbon atoms with Y double bonds in the cis configuration at position Z counted from the carboxy terminus
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