Metabolisches Engineering von Aktinomyzeten

Die Entwicklung von Antibiotikaresistenzen ist ein zunehmendes Problem bei der Therapie von Infektionskrankheiten. Die Entwicklung neuer antibiotischer Medikamente basiert häufig auf bereits bekannten Molekülen und Wirkprinzipien, so dass die Bakterien sich schnell anpassen können. Aus diesem Grund, suchen Wissenschaftler nach ganz neuen Wirkstoffen. Am Helmholtz-Institut für Pharmazeutische Forschung Saarland (HIPS), einer Außenstelle des HZI, entwickeln sie neue Wege, mit denen sie Aktinomyceten bislang unbekannte Stoffe entlocken.

Publikationen

  • Brötz, E., Bilyk, O., Kröger, S., Paululat, T., Bechthold, A., Luzhetskyy, A. (2014)Amycomycins C and D, new angucyclines from Kitasatospora sp. Tetrahedron Letters. 55(42): 5771–5773.
  • Rebets, Y., Brötz, E., Manderscheid, N., Tokovenko, B., Myronovskyi, M., Metz, P., Petzke, L., Luzhetskyy, A. (2014) Insights into the Pamamycin Biosynthesis. Angew Chem Int Ed Engl. 54 (7): 2280‐4.
  • Rebets, Y., Tokovenko, B., Lushchyk, I., Rückert, C., Zaburannyi, N., Bechthold, A., Kalinowski, J., Luzhetskyy, A. (2014) Complete genome sequence of producer of the glycopeptide antibiotic Aculeximycin Kutzneria albida DSM 43870T, a representative of minor genus of Pseudonocardiaceae. BMC Genomics. 15: 885.
  • Myronovskyi, M., Rosenkränzer, B., Luzhetskyy, A. (2014) Iterative marker excision system. Appl Microbiol Biotechnol. 98 (10): 4557‐70.
  • Horbal, L., Fedorenko, V., Luzhetskyy, A. (2014) Novel and tightly regulated resorcinol and cumate‐inducible expression systems for Streptomyces and other actinobacteria. Appl Microbiol Biotechnol. 98 (20): 8641‐55.
  • Bilyk, B. and Luzhetskyy A. (2014) Unusual site‐specific DNA integration into the highly active pseudo‐attB of the Streptomyces albus J1074 genome Appl Microbiol Biotechnol. 98(11): 5095‐104.Zaburannyi, N., Rabyk, M., Ostash, B., Fedorenko, V. and Luzhetskyy, A. (2014) Insights into naturally minimised Streptomyces albus J1074 genome BMC Genomics, 15 (1): 97.
  • Myronovskyi, M., Tokovenko, B., Brötz, E., Rückert, C., Kalinowski, J. and Luzhetskyy, A. (2014) Genome rearrangements of Streptomyces albus J1074 lead to the carotenoid gene cluster activation Appl. Microbiol. Biotechnol., 98 (2): 795‐806.Rebets, Y., Brötz, E., Tokovenko, B. and Luzhetskyy, A. (2014) Actinomycetes biosynthetic potential: how to bridge in silico and in vivo? Journal of Industrial Microbiology & Biotechnology, 41 (2): 387‐402.
  • Siegl, T., Tokovenko, B., Myronovskyi, M., and Luzhetskyy, A. (2013) Design, constructionand characterisation of a synthetic promoter library for fine‐tuned gene expression inactinomycetes. Metabolic Engineering, 19: 98‐106.Myronovskyi, M., Tokovenko, B., Manderscheid, N., Petzke, L., and Luzhetskyy, A. (2013)Complete genome sequence of Streptomyces fulvissimus. Journal of Biotechnology,168(1):117‐118.
  • Myronovskyi, M and Luzhetskyy, A. (2013) Genome engineering in actinomycetes using site‐specific recombinases. Applied Microbiology and Biotechnology, 97 (11): 4701‐4712.
  • Bilyk, B., Weber, S., Myronovskyi, M., Bilyk, O., Petzke, L. and Luzhetskyy, A. (2013) In vivo random mutagenesis of streptomycetes using mariner‐based transposon Himar1. Applied Microbiology and Biotechnology, 97 (1): 351‐359.
  • Horbal, L., Fedorenko, V., Bechthold, A. and Luzhetskyy, A. (2013) A transposon‐based strategy to identify the regulatory gene network responsible for landomycin E biosynthesis. FEMS Microbiology Letters, 342 (2): 138‐146.
  • Siegl, T. and Luzhetskyy, A. (2012) Actinomycetes genome engineering approaches. Antonie Van Leeuwenhoek, 102 (3): 503‐516.
  • Herrmann, S., Siegl, T., Luzhetska, M., Petzke, L., Jilg, C., Welle, E., Erb, A., Leadlay, P.F., Bechthold, A. and Luzhetskyy, A. (2012) Site‐specific recombination strategies for engineering actinomycete genomes. Applied and Environmental Microbiology, 78 (6): 1804‐1812.
  • Myronovskyi, M., Welle, E., Fedorenko, V. and Luzhetskyy, A. (2011) Beta‐glucuronidase as a sensitive and versatile reporter in actinomycetes. Applied and Environmental Microbiology, 77 (15): 5370‐5383.
  • Siegl, T., Petzke, L., Welle, E., and Luzhetskyy, A. (2010) I‐SceI endonuclease: a new tool for DNA repair studies and genetic manipulations in streptomycetes. Applied Microbiology and Biotechnology, 87 (4): 1525‐1532.
  • Petzke, L. and Luzhetskyy, A. (2009) In vivo Tn5‐based transposon mutagenesis of Streptomycetes. Applied Microbiology and Biotechnology, 83 (5): 979‐986.
  • Fedoryshyn, M., Petzke, L., Welle, E., Bechthold, A. and Luzhetskyy, A. (2008) Marker removal from actinomycetes genome using Flp recombinase. Gene, 419 (1‐2): 43‐47.
  • Fedoryshyn, M., Welle, E., Bechthold, A. and Luzhetskyy, A. (2008) Functional expression of the cre recombinase in actinomycetes. Applied Microbiology and Biotechnology, 78 (6): 1065‐1070.
  • Luzhetskyy, A., Hoffman, J., Pelzer, S., Wohlert, S., Vente, A. and Bechthold, A. (2008) Aranciamycin analogues generated by combinatorial biosynthesis show improved antitumor activity. Applied Microbiology and Biotechnology, 80(1): 15‐19.
  • Luzhetskyy, A., Weiss, H., Charge, A., Welle, E., Linennbrink, A. and Bechthold, A. (2007) A strategy for cloning glycosyltransferase genes involved in natural product biosynthesis. Applied Microbiology and Biotechnology, 75 (6): 1367‐1375.
  • Luzhetskyy, A., Mayer, A., Hoffmann, J., Pelzer, S., Wohlert, S., Vente, A. and Bechthold, A. (2007) Cloning and heterologous expression of the aranciamycin biosynthetic gene cluster revealed a new flexible glycosyltransferase. ChemBioChem, 8 (6): 599‐602.
  • Luzhetskyy, A., Fedoryshyn M., Gromyko O., Ostash B., Rebets Y., Bechthold A. and Fedorenko V. (2006) IncP plasmids are most effective in mediating conjugation between Escherichia coli and Streptomycetes. Russian Journal of Genetics, 42 (5): 476‐481.
  • Luzhetskyy, A. and Bechthold, A. (2005) It works: combinatorial biosynthesis for generatingnovel glycosylated compounds. Molecular Microbiology, 58 (1): 3‐5.
  • Luzhetskyy, A., Taguchi, T., Fedoryshyn, M., Dürr, C., Wohlert, S.E., Novikov, V. and Bechthold, A. (2005) LanGT2 catalyzes the first glycosylation step during Landomycin A biosynthesis. ChemBioChem, 6 (8): 1406‐1410.
  • Luzhetskyy, A., Fedoryshyn, M., Dürr, C., Taguchi, T., Novikov, V. and Bechthold, A. (2005) Iteratively acting glycosyltransferases involved in the hexasaccharide biosynthesis of Landomycin A. Chemistry & Biology, 12 (7): 725‐729.
  • Luzhetskyy, A., Zhu, L., Gibson, M., Fedoryshyn, M., Dürr, C., Hofmann, C., Hoffmeister, D.,Ostash, B., Mattingly, C., Adams, V., Fedorenko, V., Rohr, J. and Bechthold, A. (2005) Generation of novel Landomycins M and O through targeted gene disruption. ChemBioChem, 6 (4): 675‐678.
  • Luzhetskyy, A., Liu, T., Fedoryshyn, M., Ostash, B., Fedorenko, V., Rohr, J. and Bechthold, A. (2004) Function of lanGT3, a glycosyltransferase gene involved in Landomycin A biosynthesis. ChemBioChem, 5 (11): 1567‐1570.
  • Luzhetskyy, A., Fedoryshyn, M., Hoffmeister, D., Bechthold, A. and Fedorenko, V. (2002) Gene cloning system for Streptomyces cyanogenus S136. Visn.Lviv.Univ., 29, 62‐68.
  • Luzhetskyy, A., Ostash B.E. and Fedorenko V.A. (2001) Intergeneric Conjugation Escherichia coli–Streptomyces globisporus 1912 using integrative plasmid pSET152 and its derivatives / Interspecies conjugation of Escherichia coli‐Streptomyces globisporus 1912 using integrative plasmid pSET152 and its derivatives. Russian Journal of Genetics / Genetika, 37 (10): 1123‐1129 / 1340‐1347.

 

Reviews and Book Chapters

  • Luzhetskyy, A., Weitnauer, G., Bechthold, A. (2012) Prokaryotic Cells in Biotech Production. In: Kayer, O. and Warzecha, H. (Eds) Pharmaceutical Biotechnology: Pharmaceutical Biotechnology: Drug Discovery and Clinical Applications, Second Edition, Wiley-VCH Verlag GmbH & Co. KGaA Weinheim, Germany, chapter 2: 15-41.
  • Luzhetskyy, A., Pelzer, S., Bechthold, A. (2007) The future of natural products as a source of new antibiotics. Curr Opin Investig Drugs. 8 (8): 608‐613.
  • Luzhetskyy, A., Vente, A, Bechthold, A. (2005) Glycosyltransferases involved in the biosynthesis of biologically active natural products that contain oligosaccharides. Mol.Biosyst. 1 (2): 117‐126.
  • Luzhetskyy, A., Mendez, C., Salas, J.A., Bechthold, (2008) A. Glycosyltransferases, important tools for drug design. Curr Topics Med Chem. 8 (8): 680‐709.
  • Luzhetskyy, A. and Bechthold, A. (2008) Features and applications of bacterial glycosyltransferases: current state and prospects. Appl Microbiol Biotechnol. 80 (6): 945‐952.
  • Hertweck, C., Luzhetskyy, A., Rebets, Y., Bechthold, A. (2007) Type II polyketide synthases: gaining a deeper insight into enzymatic teamwork. Nap Prod Rep. 24 (1): 162‐190.
  • Mendez, C., Luzhetskyy, A., Bechthold, A., Salas, J.A. (2008) Deoxysugars in bioactivenatural products: development of novel derivatives by altering the sugar pattern. Curr Topics Med Chem. 8 (8): 710‐724.
  • Bechthold, A., Weitnauer, G., Luzhetskyy, A., Berner, M., Bihlmeier, C., Boll, R., Durr, C., Frerich, A., Hofmann, C., Mayer, A., Treede, I., Vente, A., Luzhetska, M. (2005) Glycosyltransferases and other tailoring enzymes as tools for the generation of novel compounds. Ernst Schering Research Foundation Workshop. 51: 147‐163.

 

Selected publications as a co‐author

  • Horbal, L., Kobylyanskyy, A., Truman, A.W., Zaburranyi, N., Ostash, B., Luzhetskyy, A.,Marinelli, F., Fedorenko, V. (2014) The pathway‐specific regulatory genes, tei15* and tei16*, are the master switches of teicoplanin production in Actinoplanes teichomyceticus. Appl Microbiol Biotechnol. 98 (22): 9295‐309.
  • Lopatniuk, M., Ostash, B., Luzhetskyy, A., Walker, S., Fedorenko, V. (2014) Generation and study of the strains of streptomycetes ‐ heterologous hosts for production of moenomycin. Russ J Genet. 50 (4): 360‐365.
  • Raju, R., Gromyko, O., Andriy, B., Fedorenko, V., Luzhetskyy, A., Müller, R. (2014) Oleamycins A and B: new antibacterial cyclic hexadepsipeptides isolated from a terrestrial Streptomyces sp. J Antibiot (Tokyo) in print.
  • Ostash, B., Campbell, J., Luzhetskyy, A. and Walker, S. (2013) MoeH5: a natural glycorandomizer from the moenomycin biosynthetic pathway. Mol Microbiol. 90 (6):1324‐1338.
  • Horbal, L., Kobylyanskyy, A., Yushchuk, O., Zaburannyi, N., Luzhetskyy, A., Ostash, B., Marinelli, F. and Fedorenko, V. (2013) Evaluation of heterologous promoters for genetic analysis of Actinoplanes teichomyceticus‐Producer of teicoplanin, drug of last defense. J Biotechnol. 168 (4):367‐372.
  • Raju, R., Gromyko, O., Fedorenke,V., Luzhetskyy,A. and Müller, R. (2013) Lorneic acids C and D, new trialkyl‐substituted aromatic acids isolated from a terrestrial Streptomyces sp. The Journal of Antibiotics (Tokyo), 66 (6): 347‐349.
  • Makitrynskyy, R., Ostash, B., Tsypik, O., Rebets, Y., Doud, E., Meredith, T., Luzhetskyy, A., Bechthold, A., Walker, S. and Fedorenko, V. (2013) Pleiotropic regulatory genes bldA, adpA and absB are implicated in production of phosphoglycolipid antibiotic moenomycin. Open Biol. 3 (10): 130121.
  • Raju, R., Gromyko, O., Fedorenko, V., Herrmann, J., Luzhetskyy, A. and Müller, R. (2013)Rubimycinone A, a new anthraquinone from a terrestrial Streptomyces sp. Tetrahedron Letters, 54 (8): 900‐902.
  • Raju R., Gromyko O., Fedorenko V., Luzhetskyy, A. and Müller R. (2013) Oleaceran: A Novel Spiro [isobenzofuran‐1,2'‐naptho[1,8‐bc]furan] Isolated from a Terrestrial Streptomyces sp. Organic Letters, 15 (14): 3487‐3489.
  • Strobel, T., Schmidt, Y., Linnenbrink, A., Luzhetskyy, A., Luzhetska, M., Taguchi, T., Brötz, E., Paululat, T., Stasevych, M., Stanko, O., Novikov, V. and Bechthold, A. (2013) Tracking down biotransformation to the genetic level: Identfication of a highly flexible glycosyltransferase from Saccharothrix espanaensis. Appl. and Environm. Microbiology, 79 (17): 5224‐5232.
  • Raju, R., Gromyko, O., Fedorenko, V., Luzhetskyy, A., Plaza, A. and Müller, R. (2012) Juniperolide A: A new polyketide isolated from a terrestrial actinomycete, Streptomyces sp. Organic Letters, 14 (23): 5860‐5863.
  • Raju, R., Gromyko, O., Fedorenko, V., Luzhetskyy, A. and Müller, R. (2012) Leopolic acid A, isolated from a terrestrial actinomycete, Streptomyces sp., Tetrahedron Letters 53 (14): 6300‐6301.
  • Raju, R, Gromyko, O., Fedorenko, O., Luzhetskyy, A. and Müller, R. (2012) Pimprinols A–C, from the terrestrial actinomycete, Streptomyces sp. Tetrahedron Letters, 53 (24): 3009‐3011.
  • Horbal, L., Rebets, Y., Rabyk, M., Makitrynskyy, R., Luzhetskyy, A., Fedorenko, V. and Bechthold, A. (2012) SimReg1 is a master switch for biosynthesis and export of simocyclinone D8 and its precursors. AMB Express, 2 (1):1‐8.
  • Härle, J., Günther, S., Lauinger, B., Weber, M., Kammerer, B., Zechel, D.L., Luzhetskyy, A. and Bechthold, A. (2011) Rational design of an aryl‐C‐glycoside catalyst from a natural product O‐glycosyltransferase. ChemBiol, 18 (4): 520‐530.
  • Ostash, B., Ostash, I., Zhu, L., Kharel, M.K., Luzhetskyy, A., Bechthold ,A., Walker, S., Rohr, J., and Fedorenko, V. (2010) Properties of lanK‐based regulatory circuit involved in landomycin biosynthesis in Streptomyces cyanogenus S136. Genetika, 46 (5): 604‐609.
  • Horbal, L., Rebets, Y., Rabyk, M., Luzhetskyy, A., Ostash, B., Welle, E., Nakamura, T., Fedorenko, V. and Bechthold, A. (2010) Characterization and analysis of the regulatory network involved in control of lipomycin biosynthesis in Streptomyces aureofaciens Tü117. Appl. Microbiol. Biotechnol., 85 (4): 1069‐1079.
  • Erb, A., Luzhetskyy, A., Hardter, U. and Bechthold, A. (2009) Cloning and sequencing of the biosynthetic gene cluster for saquayamycin Z and galtamycin B and elucidating the assembly of their saccharide chains. ChemBioChem, 10 (8): 1392‐1401
  • Krauth, C., Fedoryshyn, M., Schleberger, C., Luzhetskyy, A. and Bechthold, A. (2009) Engineering a function into a glycosyltransferase. ChemBiol, 16 (1): 28‐35.
  • Erb, A., Krauth, C., Luzhetskyy, A. and Bechthold, A. (2009) Differences in the substrate specificity of glycosyltransferases involved in landomycins A and E biosynthesis. Appl. Microbiol. Biotechnol., 83 (6): 1067‐1076.
  • Ostash, I., Ostash, B., Luzhetskyy, A., Bechthold, A., Walker, S. and Fedorenko, V. (2008) Coordination of export and glycosylation of landomycins in Streptomyces cyanogenus S136. FEMS Microbiol. Lett., 285 (2): 195‐202.
  • Rebets, Y., Dutko, L., Ostash, B., Luzhetskyy, A., Kulachkovskyy, O., Yamaguchi, T., Nakamura, T., Bechthold, A. and Fedorenko, V. (2008) Function of lanI in regulation of landomycin A biosynthesis in Streptomyces cyanogenus S136 and cross‐complementation studies with Streptomyces antibiotic regulatory proteins encoding genes. Arch Microbiol.,189 (2): 111‐120.
  • Fedoryshyn, M., Nur‐e‐Alam, M., Zhu, L., Luzhetskyy, A., Rohr, J. and Bechthold, A. (2007) Surprising production of a new urdamycin derivative by S. fradiae urdQ/R. J Biotechnol., 130 (1): 32‐38.
  • Zhu, L., Luzhetskyy, A., Luzhetska, M., Mattingly, C., Adams, V., Bechthold, A. and Rohr, J. (2007) Generation of New Landomycins with Altered Saccharide Patterns through Overexpression
    of the Glycosyltransferase Gene lanGT3 in the Biosynthetic Gene Cluster of Landomycin A in Streptomyces cyanogenus S‐136. ChemBioChem, 8 (1): 83‐88.
  • Dürr, C., Schnell, H.J., Luzhetskyy, A., Murillo, R., Weber, M., Welzel, K., Vente, A. and Bechthold, A. (2006) Biosynthesis of the terpene phenalinolactone in Streptomyces sp. Tü6071: analysis of the gene cluster and generation of derivatives. ChemBiol., 13 (4): 365‐377.
  • Sianidis, G., Wohlert, S.E., Pozidis, C., Karamanou, S., Luzhetskyy, A., Vente, A. and Economou, A. (2006) Cloning, purification and characterization of a functional anthracycline glycosyltransferase. J Biotechnol., 125 (3): 425‐433.
  • Baig, I., Kharel, M., Kobylyanskyy, A., Zhu, L., Rebets, Y., Ostash, B., Luzhetskyy, A., Bechthold, A., Fedorenko, V. and Rohr, J. (2006) On the Acceptor Substrate of CGlycosyltransferase UrdGT2: Three Prejadomycin C‐Glycosides from an Engineered Mutant of Streptomyces globisporus 1912 lndE(urdGT2). Angewandte Chemie Int. Ed., 45 (46): 7842‐7846.
  • Dutko, L., Rebets, Y., Ostash, B., Luzhetskyy, A., Bechthold, A., Nakamura, T. and Fedorenko, V. (2006) A putative proteinase gene is involved in regulation of landomycin E biosynthesis in Streptomyces globisporus1912. FEMS Microbiol Lett., 255 (2): 280‐285.
  • Ostash, B.E., Ogonyan, S.V., Luzhetskyy, A., Bechthold, A. and Fedorenko, V.A. (2005) The use of PCR for detecting genes that encode type I polyketide synthases in genomes of Actinomycetes. Russian Journal of Genetics / Genetika, 41 (5): 473‐478 / 2005: 41(5): 595‐
    600.
  • Mayer, A., Taguchi, T., Linnenbrink, A., Hofmann, C., Luzhetskyy, A. and Bechthold, A. (2005) LanV, a bifunctional enzyme: aromatase and ketoreductase during landomycin A biosynthesis, Chembiochem, 6 (12): 2312‐2315.
  • Rebets, Y, Ostash, B, Luzhetskyy, A., Kushnir, S, Fukuhara, M, Bechthold, A, Nashimoto, M, Nakamura, T and Fedorenko, V. (2005) DNA‐binding activity of LndI protein and temporal expression of the gene that upregulates landomycin E production in Streptomyces globisporus 1912. Microbiology, 151 (1): 281‐290.
  • Zhu, L., Ostash, B., Rix, U., Nur‐E‐Alam, M., Mayer, A., Luzhetskyy, A., Mendez, C., Salas, J.A., Bechthold, A., Fedorenko, V. and Rohr, J. (2005) Identification of the function of gene lndM2 encoding a bifunctional oxygenase‐reductase involved in the biosynthesis of the antitumor antibiotic landomycin E by Streptomyces globisporus 1912 supports the
    originally assigned structure for Landomycinone. Journal of Organic Chemistry, 70 (2): 631‐638.
  • Gromyko, O., Rebets, Y., Ostash, B., Luzhetskyy, A., Fukuhara, M., Bechthold, A., Nakamura, T. and Fedorenko, V. (2004) Generation of Streptomyces globisporus SMY622 strain with increased landomycin E production and it's initial characterization. J Antibiotics, 57 (6): 383‐389.
  • Ostash, B., Rix, U., Rix, L., Liu, T., Lombo, F., Luzhetskyy, A., Gromyko, O., Wang, C., Brana, A., Mendez, C., Salas, J.A., Fedorenko, V. and Rohr, J. (2004) Generation of new landomycins by combinatorial biosynthetic manipulation of the LndGT4 gene of the landomycin E cluster in S. globisporus. Chem Biol., 11 (4): 547‐555.
  • Mulert, U., Luzhetskyy, A., Hofmann, C., Mayer, A. and Bechthold, A. (2004) Expression of the landomycin biosynthetic gene cluster in a PKS mutant of Streptomyces fradiae is dependent on the coexpression of a putative transcriptional activator gene. FEMS Microbiol Lett., 230 (1):91‐97.
  • Rebets, Y., Ostash, B., Luzhetskyy, A., Hoffmeister, D., Brana, A., Mendez, C., Salas, J., Bechthold, A. and Fedorenko, V. (2003) Production of landomycins in Streptomyces globisporus 1912 and S cyanogenus S136 is regulated by genes encoding putative transcriptional activators. FEMS Microbiol Lett., 222 (1): 149‐153.

Patents

  • Europäische Patentanmeldung Nr.07.017025.3 “ Herstellung von genetisch modifizierten Aktinomyceten durch Rekombination”.
  • Ukrainian national patent Nr:62200A „Obtaining method of Streptomyces globisporus 1912 landomycin E overproducers”. 

 

 

Leitung

  • Dr. Andriy Luzhetskyy

    Andriy Luzhetskyy

    Leiter der Nachwuchsgruppe Metabolisches Engineering von Aktinomyzeten

    0681 302-70215

    Kontakt

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