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Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering

Paper ID Volume ID Publish Year Pages File Format Full-Text
16946 42625 2015 8 PDF Available
Title
Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering
Abstract

•The salvage pathway for GDP-fucose biosynthesis from Bacterioides fragilis was introduced into Escherichia coli.•The biosynthesis of guanosine 5′-triphosphate (GTP), an essential substrate for GDP-fucose biosynthesis, was enhanced.•The production capacities of metabolically engineered strains bearing different combinations of recombinant enzymes were compared.•The recombinant strain containing enzymes Fkp, Gpt, Gmk and Ndk have a better productivity.•The established schemes in this work also have the advantage to be a potential avenue to GDP-fucose analogs.

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.

Keywords
Escherichia coli; Salvage pathway; GDP-fucose; Guanosine nucleotides; Metabolic engineering
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Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 69, February 2015, Pages 38–45
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us