Abstract
The rifamycin gene cluster in Amycolatopsis mediterranei includes the gene rifP, whose role in antibiotic production has not yet been established. In this work, the rifP gene was silenced and the results indicated that it codes for a protein to export rifamycin, avoiding its accumulation inside the cell. An antisense cassette was constructed by inserting the rifP gene in an antisense orientation downstream from the modified ermE* promoter, and upstream of the Tasd terminator (aspartate semialdehyde dehydrogenase of A. lactamdurans). Partial silencing of the rifP gene by the use of the antisense cassette, cloned in the plasmid pUAMAE5, resulted in a 70% decrease in the extracellular rifamycin B. A protein of 53 kDa was absent in the membrane fraction of the silenced strain. This is the same size of the expected product from the rifP gene. The 2D structure analysis indicated it belongs to a Drug:H+ antiporter family which includes a wide number of membrane transport proteins.
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Acknowledgments
The authors thank to Professor Heinz G. Floss (Department of Chemistry, University of Washington, Seattle U.S.A.) for providing A. mediterranei S699 strain. This work was supported by Consejo Nacional de Ciencia y Tecnología (CONACyT, México.) (Grant SEP-2003-C02-44911).
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Absalón, A.E., Fernández, F.J., Olivares, P.X. et al. RifP; a membrane protein involved in rifamycin export in Amycolatopsis mediterranei . Biotechnol Lett 29, 951–958 (2007). https://doi.org/10.1007/s10529-007-9340-7
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DOI: https://doi.org/10.1007/s10529-007-9340-7