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Consolidated plasmid Design for Stabilized Heterologous Production of the complex natural product Siderophore Yersiniabactin |
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| Authors: | Ruiquan Qi Girish Swayambhu Michael Bruno Guojian Zhang Blaine A Pfeifer |
| Institution: | 1. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, New York, USA
Contribution: Data curation, Formal analysis, Methodology, Writing - original draft;2. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, New York, USA Contribution: Data curation, Writing - original draft;3. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, New York, USA Contribution: Data curation;4. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, New York, USA Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, People's Republic of China Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao, People's Republic of China Contribution: Methodology, Supervision;5. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, New York, USA |
| Abstract: | Yersiniabactin (Ybt) is a hybrid polyketide-nonribosomal complex natural product also known as a siderophore for its iron chelation properties. The native producer of Ybt, Yersinia pestis, is a priority pathogen responsible for the plague in which the siderophore properties of Ybt are used to sequester iron and other metal species upon host infection. Alternatively, the high metal binding properties of Ybt enable a plethora of potentially valuable applications benefiting from metal remediation and/or recovery. For these applications, a surrogate production source is highly preferred relative to the pathogenic native host. In this work, we present a modification to the heterologous Escherichia coli production system established for Ybt biosynthesis. In particular, the multiple plasmids originally used to express the genetic pathway required for Ybt biosynthesis were consolidated to a single, copy-amplifiable plasmid. In so doing, plasmid stability was improved from ~30% to ≥80% while production values maintained at 20–30% of the original system, which resulted in titers of 0.5–3 mg/L from shake flask vessels. |
| Keywords: | E coli nonribosomal peptide polyketide siderophore Yersiniabactin |