Reconstruction of the archaeal isoprenoid ether lipid biosynthesis pathway in Escherichia coli through digeranylgeranylglyceryl phosphate |
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| Authors: | Denton Lai Ben Lluncor Imke Schröder Robert P Gunsalus James C Liao Harold G Monbouquette |
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| Institution: | 1. Chemical and Biomolecular Engineering Department, University of California, Box 951592, Los Angeles, CA 90095-1592, USA;2. Microbiology, Immunology, and Molecular Genetics Department, University of California, Los Angeles, CA 90095, USA;1. Department of Life Sciences, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea;2. Department of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea;3. School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea;4. Department of Chemical and Biological Engineering, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul 02841, Korea;1. Department of Marine Organic Biogeochemistry, Royal NIOZ Netherlands Institute for Sea Research, P.O. Box 59, 1790AB Den Burg, Texel, The Netherlands;2. Department of Marine Geology, Royal NIOZ Netherlands Institute for Sea Research, Texel, The Netherlands;3. MARUM-Center for Marine Environmental Sciences, MARUM – Center for Marine Environmental Sciences, University of Bremen, Leobener Str., D-28359 Bremen, Germany;1. Alfred-Wegener-Institut Helmholtz-Zentrum für Polar- und Meeresforschung (AWI), D-27570 Bremerhaven, Germany;2. MARUM Center for Marine Environmental Sciences, Department of Geosciences, University of Bremen, D-28359 Bremen, Germany;1. Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, Los Angeles, CA 90095, United States;2. Academia Sinica, Taipei 11529, Taiwan;1. Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan;2. Department of Biomolecular Chemistry, Kyoto Prefectural University, Hangi-cho, Shimogamo, Sakyo-ku, Kyoto 606-8522, Japan |
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| Abstract: | The membrane lipids of archaea are characterized by unique isoprenoid biochemistry, which typically is based on two core lipid structures, sn-2,3-diphytanylglycerol diether (archaeol) and sn-2,3-dibiphytanyldiglycerol tetraether (caldarchaeol). The biosynthetic pathway for the tetraether lipid entails unprecedented head-to-head coupling of isoprenoid intermediates by an unknown mechanism involving unidentified enzymes. To investigate the isoprenoid ether lipid biosynthesis pathway of the hyperthermophilic archaeon, Archaeoglobus fulgidus, its lipid synthesis machinery was reconstructed in an engineered Escherichia coli strain in an effort to demonstrate, for the first time, efficient isoprenoid ether lipid biosynthesis for the production of the intermediate, digeranylgeranylglyceryl phosphate (DGGGP). The biosynthesis of DGGGP was verified using an LC/MS/MS technique and was accomplished by cloning and expressing the native E. coli gene for isopentenyl diphosphate (IPP) isomerase (idi), along with the A. fulgidus genes for G1P dehydrogenase (egsA) and GGPP synthase (gps), under the control of the lac promoter. The A. fulgidus genes for GGGP synthase (GGGPS) and DGGGP synthase (DGGGPS), under the control of the araBAD promoter, were then introduced and expressed to enable DGGGP biosynthesis in vivo. This investigation established roles for four A. fulgidus genes in the isoprenoid ether lipid pathway for DGGGP biosynthesis and provides a platform useful for identification of subsequent, currently unknown, steps in tetraether lipid biosynthesis proceeding from DGGGP, which is the presumed substrate for the head-to-head coupling reaction yielding unsaturated caldarchaeol. |
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