TREX: A Universal Tool for the Transfer and Expression of Biosynthetic Pathways in Bacteria

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• 作者：Anita Loeschcke ; Annette Markert ; Susanne Wilhelm ; Astrid Wirtz ; Frank Rosenau ; Karl-Erich Jaeger ; Thomas Drepper
• 刊名：ACS Synthetic Biology
• 出版年：2013
• 出版时间：January 18, 2013
• 年：2013
• 卷：2
• 期：1
• 页码：22-33
• 全文大小：590K
• 年卷期：v.2,no.1(January 18, 2013)
• ISSN：2161-5063

文摘

Secondary metabolites represent a virtually inexhaustible source of natural molecules exhibiting a high potential as pharmaceuticals or chemical building blocks. To gain broad access to these compounds, sophisticated expression systems are needed that facilitate the transfer and expression of large chromosomal regions, whose genes encode complex metabolic pathways. Here, we report on the development of the novel system for the transfer and expression of biosynthetic pathways (TREX), which comprises all functional elements necessary for the delivery and concerted expression of clustered pathway genes in different bacteria. TREX employs (i) conjugation for DNA transfer, (ii) randomized transposition for its chromosomal insertion, and (iii) T7 RNA polymerase for unimpeded bidirectional gene expression. The applicability of the TREX system was demonstrated by establishing the biosynthetic pathways of two pigmented secondary metabolites, zeaxanthin and prodigiosin, in bacteria with different metabolic capacities. Thus, TREX represents a valuable tool for accessing natural products by allowing comparative expression studies with clustered genes.

Keywords:

transfer of biosynthetic pathways; functional expression of clustered genes; secondary metabolites discovery; synthetic biology; metabolic engineering