Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering |
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| Institution: | 1. National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, People''s Republic of China;2. The State Key Laboratory of Microbial Technology and School of Life Science, Shandong University, Jinan, Shandong 250100, People''s Republic of China;1. Institute of Microbiology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany;2. Bioorganic Chemistry, Institute of Chemistry, University of Hohenheim, Garbenstrasse 30, 70599 Stuttgart, Germany;1. Graz University of Technology, Institute of Biotechnology and Biochemical Engineering, NAWI Graz, Petersgasse 12, A-8010 Graz, Austria;2. Austrian Centre of Industrial Biotechnology, Petersgasse 14, A-8010 Graz, Austria;1. Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 151-921, Republic of Korea;2. Department of Bioenergy Science and Technology, Chonnam National University, Gwangju 500-757, Republic of Korea;1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People’s Republic of China;2. Bloomage Biotechnology Corp., Ltd., Jinan, Shandong 250010, People’s Republic of China |
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| Abstract: | Guanosine 5′-diphosphate (GDP)-fucose is the indispensible donor substrate for fucosyltransferase-catalyzed synthesis of fucose-containing biomolecules, which have been found involving in various biological functions. In this work, the salvage pathway for GDP-fucose biosynthesis from Bacterioides fragilis was introduced into Escherichia coli. Besides, the biosynthesis of guanosine 5′-triphosphate (GTP), an essential substrate for GDP-fucose biosynthesis, was enhanced via overexpression of enzymes involved in the salvage pathway of GTP biosynthesis. The production capacities of metabolically engineered strains bearing different combinations of recombinant enzymes were compared. The shake flask fermentation of the strain expressing Fkp, Gpt, Gmk and Ndk obtained the maximum GDP-fucose content of 4.6 ± 0.22 μmol/g (dry cell mass), which is 4.2 fold that of the strain only expressing Fkp. Through fed-batch fermentation, the GDP-fucose content further rose to 6.6 ± 0.14 μmol/g (dry cell mass). In addition to a better productivity than previous fermentation processes based on the de novo pathway for GDP-fucose biosynthesis, the established schemes in this work also have the advantage to be a potential avenue to GDP-fucose analogs encompassing chemical modification on the fucose residue. |
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| Keywords: | Salvage pathway GDP-fucose Guanosine nucleotides Metabolic engineering |
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