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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
Affiliation: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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