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Consolidated plasmid Design for Stabilized Heterologous Production of the complex natural product Siderophore Yersiniabactin
Biotechnology Progress ( IF 2.9 ) Pub Date : 2020-11-14 , DOI: 10.1002/btpr.3103
Ruiquan Qi 1 , Girish Swayambhu 1 , Michael Bruno 1 , Guojian Zhang 1, 2, 3 , Blaine A Pfeifer 1, 2, 3
Affiliation  

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.

中文翻译:

用于稳定异源生产复杂天然产物铁载体耶尔森菌素的整合质粒设计

耶尔森菌素 (Ybt) 是一种混合聚酮化合物-非核糖体复合物天然产物,因其铁螯合特性而被称为铁载体。Ybt 的本地生产者鼠疫耶尔森氏菌是导致瘟疫的主要病原体,其中 Ybt 的铁载体特性用于在宿主感染后隔离铁和其他金属物种。或者,Ybt 的高金属结合特性使大量潜在有价值的应用受益于金属修复和/或回收。对于这些应用,替代生产来源相对于致病性天然宿主是高度优选的。在这项工作中,我们对异源大肠杆菌进行了修改建立Ybt生物合成生产体系。特别是,最初用于表达 Ybt 生物合成所需的遗传途径的多个质粒被整合到一个单一的、可复制的质粒中。这样做时,质粒稳定性从约 30% 提高到≥80%,而生产值保持在原始系统的 20-30%,这导致摇瓶容器的滴度为 0.5-3 mg/L。
更新日期:2020-11-14
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