title: Dihydroxyacetone production in an engineered Escherichia coli through expression of Corynebacterium glutamicum dihydroxyacetone phosphate dephosphorylase.
abstract: Dihydroxyacetone (DHA) has several industrial applications such as a tanning agent in tanning lotions in the cosmetic industry; its production via microbial fermentation would present a more sustainable option for the future. Here we genetically engineered Escherichia coli (E. coli) for DHA production from glucose. Deletion of E. coli triose phosphate isomerase (tpiA) gene was carried out to accumulate dihydroxyacetone phosphate (DHAP), for use as the main intermediate or precursor for DHA production. The accumulated DHAP was then converted to DHA through the heterologous expression of Corynebacterium glutamicum DHAP dephosphorylase (cghdpA) gene. To conserve DHAP exclusively for DHA production we removed methylglyoxal synthase (mgsA) gene in the ΔtpiA strain. This drastically improved DHA production from 0.83g/l (0.06g DHA/g glucose) in the ΔtpiA strain bearing cghdpA to 5.84g/l (0.41g DHA/g glucose) in the ΔtpiAΔmgsA double mutant containing the same gene. To limit the conversion of intracellular DHA to glycerol, glycerol dehydrogenase (gldA) gene was further knocked out resulting in a ΔtpiAΔmgsAΔgldA triple mutant. This triple mutant expressing the cghdpA gene produced 6.60g/l of DHA at 87% of the maximum theoretical yield. In summary, we demonstrated an efficient system for DHA production in genetically engineered E. coli strain.
authors.affiliation: Industrial Biotechnology Division, Institute of Chemical & Engineering Sciences, Agency for Science, Technology and Research (A-STAR), 1, Pesek Road, Jurong Island 627833, Singapore. Electronic address: jainvk@ices.a-star.edu.sg.
authors.forename: Vishist Kumar
authors.initials: VK
authors.lastname: Jain
authors.affiliation: Industrial Biotechnology Division, Institute of Chemical & Engineering Sciences, Agency for Science, Technology and Research (A-STAR), 1, Pesek Road, Jurong Island 627833, Singapore.
authors.forename: Crystal Jing Ying
authors.initials: CJ
authors.lastname: Tear
authors.affiliation: Industrial Biotechnology Division, Institute of Chemical & Engineering Sciences, Agency for Science, Technology and Research (A-STAR), 1, Pesek Road, Jurong Island 627833, Singapore.
authors.forename: Chan Yuen
authors.initials: CY
authors.lastname: Lim
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics; D004098:Dihydroxyacetone / Q000096:biosynthesis*; D004099:Dihydroxyacetone Phosphate / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D017353:Gene Deletion; D007700:Kinetics; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways
chemical_list: D001426:Bacterial Proteins; D004099:Dihydroxyacetone Phosphate; D004098:Dihydroxyacetone
keywords: Corynebacterium glutamicum; Dihydroxyacetone; Dihydroxyacetone phosphate; Dihydroxyacetone phosphate dephosphorylase; Escherichia coli; Metabolic engineering