title: Butyrate production in engineered Escherichia coli with synthetic scaffolds.
abstract: Butyrate pathway was constructed in recombinant Escherichia coli using the genes from Clostridium acetobutylicum and Treponema denticola. However, the pathway constructed from exogenous enzymes did not efficiently convert carbon flux to butyrate. Three steps of the productivity enhancement were attempted in this study. First, pathway engineering to delete metabolic pathways to by-products successfully improved the butyrate production. Second, synthetic scaffold protein that spatially co-localizes enzymes was introduced to improve the efficiency of the heterologous pathway enzymes, resulting in threefold improvement in butyrate production. Finally, further optimizations of inducer concentrations and pH adjustment were tried. The final titer of butyrate was 4.3 and 7.2 g/L under batch and fed-batch cultivation, respectively. This study demonstrated the importance of synthetic scaffold protein as a useful tool for optimization of heterologous butyrate pathway in E. coli.
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Sungbuk-Gu, Seoul 136-713, South Korea.
authors.forename: Jang-Mi
authors.initials: JM
authors.lastname: Baek
authors.forename: Suman
authors.initials: S
authors.lastname: Mazumdar
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Lee
authors.forename: Moo-Young
authors.initials: MY
authors.lastname: Jung
authors.forename: Jae-Hyung
authors.initials: JH
authors.lastname: Lim
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Seo
authors.forename: Gyoo-Yeol
authors.initials: GY
authors.lastname: Jung
authors.forename: Min-Kyu
authors.initials: MK
authors.lastname: Oh
mesh_terms: D000085:Acetates / Q000032:analysis / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002087:Butyrates / Q000032:analysis / Q000378:metabolism; D046969:Clostridium acetobutylicum / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000235:genetics / Q000502:physiology; D058615:Synthetic Biology / Q000379:methods*; D048373:Treponema denticola / Q000201:enzymology / Q000235:genetics
chemical_list: D000085:Acetates; D001426:Bacterial Proteins; D002087:Butyrates; D005947:Glucose
keywords: Escherichia coli; butyrate; heterologous pathway; metabolic engineering; synthetic scaffold