title: Production of 3-hydroxypropionic acid via malonyl-CoA pathway using recombinant Escherichia coli strains.
abstract: Malonyl-CoA is an intermediary compound that is produced during fatty acid metabolism. Our study aimed to produce the commercially important platform chemical 3-hydroxypropionic acid (3-HP) from its immediate precursor malonyl-CoA by recombinant Escherichia coli strains heterologously expressing the mcr gene of Chloroflexus aurantiacus DSM 635, encoding an NADPH-dependent malonyl-CoA reductase (MCR). The recombinant E. coli overexpressing mcr under the T5 promoter showed MCR activity of 0.015 U mg⁻¹ protein in crude cell extract and produced 0.71 mmol/L of 3-HP in 24h in shake flask cultivation under aerobic conditions with glucose as the sole source of carbon. When acetyl-CoA carboxylase and biotinilase, encoded by the genes accADBCb (ACC) of E. coli K-12 were overexpressed along with MCR, the final 3-HP titer improved by 2-fold, which is 1.6 mM. Additional expression of the gene pntAB, encoding nicotinamide nucleotide transhydrogenase that converts NADH to NADPH, increased 3-HP production to 2.14 mM. The strain was further developed by deleting the sucAB gene, encoding α-ketoglutarate dehydrogenase complex in tricarboxylic acid (TCA) cycle, or blocking lactate and acetate production pathways, and evaluated for the production of 3-HP. We report on the feasibility of producing 3-HP from glucose through the malonyl-CoA pathway.
authors: Department of Chemical and Biomolecular Engineering, Pusan National University, Busan 609-735, Republic of Korea.
authors: Chelladurai
authors: C
authors: Rathnasingh
authors: Subramanian Mohan
authors: SM
authors: Raj
authors: Youjin
authors: Y
authors: Lee
authors: Christy
authors: C
authors: Catherine
authors: Somasundar
authors: S
authors: Ashok
authors: Sunghoon
authors: S
authors: Park
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000378:metabolism; D000332:Aerobiosis; D000693:Anaerobiosis; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D017353:Gene Deletion; D005947:Glucose / Q000378:metabolism; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000096:biosynthesis; D008316:Malonyl Coenzyme A / Q000378:metabolism*; D053858:Metabolic Networks and Pathways; D009250:NADP Transhydrogenases / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D013696:Temperature; D013997:Time Factors
chemical_list: D019344:Lactic Acid; D008316:Malonyl Coenzyme A; C031601:hydracrylic acid; D010088:Oxidoreductases; C454539:malonyl-Coa reductase; D009250:NADP Transhydrogenases; D000103:Acetyl-CoA Carboxylase; D005947:Glucose