Journal
ACS SYNTHETIC BIOLOGY
Volume 2, Issue 1, Pages 22-33Publisher
AMER CHEMICAL SOC
DOI: 10.1021/sb3000657
Keywords
transfer of biosynthetic pathways; functional expression of clustered genes; secondary metabolites discovery; synthetic biology; metabolic engineering
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Funding
- Federal Ministry of Education and Research [GenoMik-Plus FKZ 0313751J, ExpresSys FKZ 0315586B]
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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 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 (ill) T7 RNA polymerase for unimpeded bidirectional 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.
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