RT Journal Article
SR Electronic
T1 Prototyping a valinomycin biosynthesis pathway within a cell-free transcription-translation (TX-TL) system
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 091520
DO 10.1101/091520
A1 Tiffany Zhou
YR 2016
UL http://biorxiv.org/content/early/2016/12/04/091520.abstract
AB Many natural metabolites have antibacterial, antiviral, or anticancer effects and can be developed into new drugs. However, working with the microorganisms that produce these products can be challenging since they are not as well characterized as a model organism like Escherichia coli. In this paper, we investigate the potential for a cell-free transcription-translation (TX-TL) system to provide a rapid prototyping platform for characterizing new genetic pathways. We use the valinomycin biosynthesis pathway as a test case, and we show successful heterologous expression of the heterodimeric valinomycin synthetase (VlmSyn, Vlm1: 374 kDa and Vlm2: 284 kDa) from Strep-tomyces tsusimaensis within the TX-TL system. Using LC-MS analysis, we find that valinomycin is produced at low but detectable levels, even when only one out of the three basic precursors is fed into the system. Our work represents another step towards applying cell-free biosynthesis to the discovery and characterization of new natural products.