Function of lanI in regulation of landomycin A biosynthesis in Streptomyces cyanogenus S136 and cross-complementation st
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ORIGINAL PAPER
Function of lanI in regulation of landomycin A biosynthesis in Streptomyces cyanogenus S136 and cross-complementation studies with Streptomyces antibiotic regulatory proteins encoding genes Yuriy Rebets · Lilia Dutko · Bohdan Ostash · Andriy Luzhetskyy · Olexandr Kulachkovskyy · Toshio Yamaguchi · Tatsunosuke Nakamura · Andreas Bechthold · Victor Fedorenko
Received: 6 February 2007 / Revised: 1 June 2007 / Accepted: 26 July 2007 / Published online: 5 September 2007 © Springer-Verlag 2007
Abstract The transcriptional regulator of landomycin A biosynthesis encoded by lanI gene has been inactivated within the chromosome of Streptomyces cyanogenus S136. The obtained mutant strain did not produce landomycin A and its known intermediates. Loss of landomycin A production caused signiWcant changes in morphology of the lanI deWcient strain. RT-PCR analysis conWrmed complete cessation of transcription of certain lan genes, including lanJ (encoding putative proton dependent transporter) and lanK (presumably involved in lanJ expression regulation). Intro-
Communicated by Jean-Luc Pernodet. Y. Rebets and L. Dutko have contributed equally to this work. Electronic supplementary material The online version of this article (doi:10.1007/s00203-007-0299-5) contains supplementary material, which is available to authorized users. Y. Rebets · L. Dutko · B. Ostash · O. Kulachkovskyy · V. Fedorenko (&) Department of Genetics and Biotechnology of Ivan Franko National University of L’viv, Grushevskogo st.4, L’viv, 79005, Ukraine e-mail: [email protected] Y. Rebets e-mail: [email protected] A. Luzhetskyy · A. Bechthold Institut für Pharmazeutische Wissenschaften, Lehrstuhl für Pharmazeutische Biologie und Biotechnologie Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany Y. Rebets · T. Yamaguchi · T. Nakamura Department of Microbiology, Niigata University of Pharmacy and Applied Life Sciences, Higashijima 265-1, 956-8603 Niigata, Japan
duction of either lanI or lndI [lanI homologue controlling landomycin E biosynthesis in Streptomyces globisporus 1912, both encoding Streptomyces antibiotic regulatory proteins (SARPs)] restored landomycin A production in the mutant strain. Chimeric constructs ladI and ladR were generated by exchanging the DNA sequences corresponding to Nand C-terminal parts of LndI and LanI. None of these genes were able to activate the production of landomycins in regulatory mutants of S. cyanogenus and S. globisporus. Nevertheless, the production of novel unidentiWed compound was observed in the case of S. cyanogenus harboring ladI gene. Various genes encoding SARPs have been expressed in S. globisporus and S. cyanogenus regulatory mutants and the results of these complementation experiments are discussed. Keywords Streptomyces · Antibiotics · Polyketide · SARP · Transcriptional regulators
Introduction EVorts in streptomycete gene cloning lead to identiWcation and studying of huge number of biosynthetic gene clusters governing the production of important sec
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