Expanding the chemical diversity of natural esters by engineering a polyketide-derived pathway into Escherichia coli.
Microbial fatty acid (FA)-derived molecules have emerged as promising alternatives to petroleum-based chemicals for reducing dependence on fossil hydrocarbons. However, native FA biosynthetic pathways often yield limited structural diversity, and therefore restricted physicochemical properties, of the end products by providing only a limited variety of usually linear hydrocarbons. Here we have engineered into Escherichia coli a mycocerosic polyketide synthase-based biosynthetic pathway from Mycobacterium tuberculosis and redefined its biological role towards the production of multi-methyl-branched-esters (MBEs) with novel chemical structures. Expression of FadD28, Mas and PapA5 enzymes enabled the biosynthesis of multi-methyl-branched-FA and their further esterification to an alcohol. The high substrate tolerance of these enzymes towards different FA and alcohol moieties resulted in the biosynthesis of a broad range of MBE. Further metabolic engineering of the MBE producer strain coupled this system to long-chain-alcohol biosynthetic pathways resulting in de novo production of branched wax esters following addition of only propionate.
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional deInvestigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000 Rosario, Argentina.
Simón
S
Menendez-Bravo
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional deInvestigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000 Rosario, Argentina.
Santiago
S
Comba
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional deInvestigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000 Rosario, Argentina.
Martín
M
Sabatini
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional deInvestigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000 Rosario, Argentina. Electronic address: arabolaza@ibr-conicet.gov.ar.
Ana
A
Arabolaza
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional deInvestigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000 Rosario, Argentina. Electronic address: gramajo@ibr-conicet.gov.ar.
Hugo
H
Gramajo
D000217:Acyltransferases / Q000096:biosynthesis / Q000235:genetics; D019730:Carbon-Sulfur Ligases / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D004952:Esters / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D060847:Metabolic Engineering; D009169:Mycobacterium tuberculosis; D061065:Polyketides / Q000378:metabolism*
D004952:Esters; D005227:Fatty Acids; D061065:Polyketides; D000217:Acyltransferases; C487811:PapA5 protein, Mycobacterium tuberculosis; D019730:Carbon-Sulfur Ligases; C510957:FadD28 protein, Mycobacterium tuberculosis
Branched fatty acid; Lipid metabolism; Metabolic engineering; Mycocerosic acid; Polyketide-derived molecules