title: Molecular variation of the nonribosomal peptide-polyketide siderophore yersiniabactin through biosynthetic and metabolic engineering.
abstract: The production of the mixed nonribosomal peptide-polyketide natural product yersiniabactin (Ybt) has been established using E. coli as a heterologous host. In this study, precursor-directed biosynthesis was used to generate five new analogs of Ybt, demonstrating the flexibility of the heterologous system and the biosynthetic process in allowing compound diversity. A combination of biosynthetic and cellular engineering was then used to influence the production metrics of the resulting analogs. First, the cellular levels and activity of FadL, a hydrocarbon transport protein, were tested for subsequent influence upon exogenous precursor uptake and Ybt analog production with a positive correlation observed between FadL over-production and analog formation. Next, a Ybt biosynthetic editing enzyme was removed from the heterologous system which decreased native compound production but increased analog formation. A final series of experiments enhanced endogenous anthranilate towards complete pathway formation of the associated analog which showed a selective ability to bind gold.
authors.affiliation: Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, New York.
authors.forename: Mahmoud Kamal
authors.initials: MK
authors.lastname: Ahmadi
authors.affiliation: Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, New York.
authors.forename: Samar
authors.initials: S
authors.lastname: Fawaz
authors.affiliation: Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, New York.
authors.forename: Lei
authors.initials: L
authors.lastname: Fang
authors.affiliation: Department of Chemistry, University at Buffalo, The State University of New York, Buffalo, New York.
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, New York. blainepf@buffalo.edu.
authors.forename: Blaine A
authors.initials: BA
authors.lastname: Pfeifer
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D053898:Biosynthetic Pathways; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D050612:Fatty Acid Transport Proteins / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D010636:Phenols / Q000737:chemistry / Q000378:metabolism*; D017262:Siderophores / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D050612:Fatty Acid Transport Proteins; D010636:Phenols; D017262:Siderophores; D013844:Thiazoles; C065691:fadL protein, E coli; C104398:yersiniabactin
keywords: E. coli; analog; nonribosomal peptide; polyketide; siderophore; yersiniabactin