Abstract
Streptomyces ghanaensis ATCC14672 is the producer of phosphoglycolipid antibiotics moenomycins that for almost 40 years were used worldwide as an animal feed additive. As the use of moenomycins narrows down (due to bans in the EU and some other countries), it opens the opportunity to develop much-needed antibiotics against Gram-positive human pathogens, such as cocci. It is desirable to develop ATCC14672 strains accumulating only certain members of moenomycin family which would facilitate their purification, analysis and/or chemical modification. Here we tested site-specific recombinases (SSRs) as a tool to manipulate the genome of ATCC14672 and to achieve aforementioned goals. We show that of three SSRs tested — Cre, Dre, and Flp — the first two efficiently catalyzed recombination reactions, while Flp showed no activity in ATCC14672 cells. Cre recombinase can be reused at least three times to modify ATCC14672 genome without detrimental effects, such as large-scale inversions or deletions. Properties of the generated strains and SSRs are discussed.
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References
Fedoryshyn M, Petzke L, Welle E, Bechthold A, Luzhetskyy A (2008) Marker removal from actinomycetes genome using Flp recombinase. Gene 419:43–47. doi:10.1016/j.gene.2008.04.011
Gampe CM, Tsukamoto H, Wang A, Walker S, Kahne D (2011) Modular synthesis of diphospholipid oligosaccharide fragments of the bacterial cell wall and their use to study the mechanism of moenomycin and other antibiotics. Tetrahedron 67:9771–9778. doi:10.1016/j.tet.2011.09.114
Makitrynskyy R, Rebets Y, Ostash B, Zaburannyi N, Rabyk M, Walker S, Fedorenko V (2010) Genetic factors that influence moenomycin production in streptomycetes. J Ind Microbiol Biotechnol 37:559–566
Makitrynskyy R, Ostash B, Tsypik O, Rebets Y, Doud E, Meredith T, Luzhetskyy A, Bechthold A, Walker S, Fedorenko V (2013) Pleiotropic regulatory genes bldA, adpA and absB are implicated in production of phosphoglycolipid antibiotic moenomycin. Open Biol 3:130121. doi:10.1098/rsob.130121
Ostash B, Walker S (2010) Moenomycin family antibiotics: chemical synthesis, biosynthesis, biological activity. Nat Prod Rep 27:1594–1617. doi:10.1039/c001461n
Ostash B, Saghatelian A, Walker S (2007) A streamlined metabolic pathway for the biosynthesis of moenomycin A. Chem Biol 14:257–267. doi:10.1016/j.chembiol.2007.01.008
Ostash B, Makitrinskyy R, Walker S, Fedorenko V (2009) Identification and characterization of Streptomyces ghanaensis ATCC14672 integration sites for three actinophage-based plasmids. Plasmid 61:171–175. doi:10.1016/j.plasmid.2008.12.002
Ostash B, Campbell J, Luzhetskyy A, Walker S (2013) MoeH5: a natural glycorandomizer from the moenomycin biosynthetic pathway. Mol Microbiol 90:1324–1338. doi:10.1111/mmi.12437
Pfaller MA (2006) Flavophospholipol use in animals: positive implications for antimicrobial resistance based on its microbiologic properties. Diagn Microbiol Infect Dis 56:115–1121. doi:10.1016/j.diagmicrobio.2006.03.014
Rebets Y, Lupoli T, Qiao Y, Schirner K, Villet R, Hooper D, Kahne D, Walker S (2014) Moenomycin resistance mutations in Staphylococcus aureus reduce peptidoglycan chain length and cause aberrant cell division. ACS Chem Biol 9:459–467. doi:10.1021/cb4006744
Schweizer HP (2003) Applications of the Saccharomyces cerevisiae Flp-FRT system in bacterial genetics. J Mol Microbiol Biotechnol 5:67–77. doi:10.1159/000069976
Siegl T, Luzhetskyy A (2012) Actinomycetes genome engineering approaches. Antonie Van Leeuwenhoek 102(3):503–516. doi:10.1007/s10482-012-9795-y
Welzel P (2005) Syntheses around the transglycosylation step in peptidoglycan biosynthesis. Chem Rev 105:4610–4660. doi:10.1021/cr040634e
Acknowledgments
This work was supported by grant Bg-98f from Ministry of Education and Science of Ukraine (to V.F.) and by NIH grant R03TW009424 (to S.W. and V.F.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. M.L. and R.M. were supported by DAAD fellowships.
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Communicated by: Agnieszka Szalewska-Palasz
M. Lopatniuk and B. Ostash contributed equally to this work.
Fedorenko holds a PhD, Ivan Franko National University of Lviv.
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Lopatniuk, M., Ostash, B., Makitrynskyy, R. et al. Testing the utility of site-specific recombinases for manipulations of genome of moenomycin producer Streptomyces ghanaensis ATCC14672. J Appl Genetics 56, 547–550 (2015). https://doi.org/10.1007/s13353-015-0283-8
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DOI: https://doi.org/10.1007/s13353-015-0283-8