Mikrobiol. Z. 2016; 78(6):60-70.
Sequences of Landomycin E and Carotenoid Biosynthetic Gene Clusters, andMolecular Structure
of Transcriptional Regulator of Streptomyces globisporus 1912
Matselyukh B.P., Polishchuk L.V., Lukyanchuk V.V., Golembiovska S.L., Lavrenchuk V.Y.
Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
Streptomyces globisporus 1912 and its derivatives 1912-2 and 1912-4Crt are the producers of the landomycin E, carotenoids and the regulator of antibiotics biosynthesis and morphogenesis of streptomycetes. The genome DNA of two mutant strains, 1912-2, the more effective producer of the landomycin E and the regulator, and 1912-4Crt, the producer of beta-carotene and lycopene, was sequenced by Illumina. Comparative analysis of the DNA sequences using GenBank data allowed localization of 36 landomycin E biosynthetic genes lnd of S. globisporus 1912 in one cluster. Twenty of these lnd genes have been sequenced for the first time. The new regulatory responce gene lndRR and the sensor kinase gene lndY1 were proposed as the members of the putative two-component system. High identity (94 – 95 %) was determined for the lnd genes of the 1912 strain and those of the metagenomic clone AZ97. Seven carotenoid biosynthetic genes crt of the strain 1912-4Crt were sequenced and localized in one cluster consisting of two convergent operons from 4 and 3 crt genes. High homology (93 %) of the crt gene clusters of S. globisporus 1912 and S. griseus IFO 13350 was shown. Two non-punctual repeats (NPRs) of 21 bp were identified in the sequence of crtY gene coding lycopene cyclase. It was shown that the deletion of 117 bp including the sequence between NPRs of 96 bp and one NPR from 5`-side activated the crt gene cluster and increased the production of beta carotene (6.91 mg/l) and lycopene (3.24 mg/l) by the strain 1912-4Crt. Deletion of 86 bp was revealed in the regulatory gene lndRR resulting in the deficiency of landomycin E production in the strain 1912-4Crt. The DNA sequences of crt and lnd genes of S. globisporus 1912 were submitted to the NCBI database with accession numbers KM349312 and KJ645792, respectively.
S. globisporus 1912 produced a low-molecular-weight compound that, like A-factor, restored the landomycin E and streptomycin biosynthesis and sporulation of the defective mutants S. globisporus 1912-B2 and S. griseus 1439, respectively. The compound was purified by thin layer chromatography and HPLC. It had an absorption maximum at λmax = 245 nm and a molecular mass m/z 244. On the basis of NMR spectroscopy the chemical structure of the transcriptional regulator was elucidated as the new (L)-N-methylphenylalanyl-dehydrobutyrine diketopiperazine.
Key words: Streptomyces globisporus, landomycin E, beta-carotene, lycopene, diketopiperazine, gene clusters, sequences.
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- Borisova LN, Ivkina NS, Rapoport IA. Chromatographic study of the mutants of Actinomyces streptomycini induced by nitrosomethylbiuret. Dokl. Akad. Nauk SSSR. 1996; 171: 782–732. Russian.
- Fedorenko V, Basiliya L, Pankevych K, Dubitska L, Ostash B, Luzhetskyy A, et al. Genetic control of biosynthesis of antitumor antibiotics-polyketides by actinomycetes. Bull. Inst. Agr. Microbiol. 2000; 8: 27–31. Ukrainian.
- Frenglova G, Breshkova D. Carotenoids from Rhodotorula and Phaffia: yeasts of biotechnological importance. J. Industr. Microbiol. Biotechnol. 2009; 36: 163–180. https://doi.org/10.1007/s10295-008-0492-9
- Gillet JP, Gottesman MM. Mechanisms of multidrug resistance in cancer. Methods Mol. Biol. 2010; 596: 47–76. https://doi.org/10.1007/978-1-60761-416-6_4
- Hara H, Ohnishi J, Horinouchi S. DNA microarray analysis of global gene regulator by A-factor in Streptomyces griseus. Microbiology 2009; 155: 2197–2210. https://doi.org/10.1099/mic.0.027862-0
- Khokhlov AS. Low-molecular weight microbial regulators. Moscow: Nauka: 1988. Russian.
- Kieser T, Bibb M, Buttner M, Chater K, Hopwood D. Practical Streptomyces Genetics. The John Innes Foundation: Norwich; 2000.
- Korynevska A, Heffeter P, Matseliukh B, Elbling L, Micksche M, Stoika R, et al. Mechanisms underlying the anticancer activities of the angucycline landomycin E. Biochem. Pharmacol. 2007; 74: 1713–1726. https://doi.org/10.1016/j.bcp.2007.08.026
- Krohn K, Rohr J. Angucyclines: total synthesis, new structures, and biosynthetic studies of an emerging new class of antibiotics. Nat. Prod. Res. 1997; 188: 127–195.
- Lee HS, Ohnishi J, Horinouchi S. A σB-like factor responsible for carotenoid biosynthesis in Streptomyces griseus. J. Mol. Microbiol. Biotechnol. 2001; 3: 95–101.
- Matselyukh B, Lutchenko V, Polishchuk L. Synthesis of carotenoids by mutant strains of Streptomyces globisporus 1912. Mikrobiol. Z. 2003; 65: 24–30. Ukrainian.
- Matselyukh BP, Tymoshenko SG, Bambura OI, Kopejko OP. Screening and characterization of regulators of landomycin E biosynthesis. Mikrobiol. Z. 2011; 73: 16–20. Ukrainian.
- Matselyukh BP, Polishchuk LV, Lukyanchuk VV, Golembiovska SL, Lavrenchuk VY. Molecular mechanism of the carotenoid biosynthesis activation in the producer Streptomyces globisporus 1912. Biotechnologia acta. 2014; 7(6): 69–74. https://doi.org/10.15407/biotech7.06.069
- Matselyukh BP, Polishchuk LV, Lukyanchuk VV. Complete sequence of landomycin E biosynthetic gene cluster from Streptomyces globisporus 1912. Mikrobiol. Z. 2015; 77(1): 51–56. https://doi.org/10.15407/microbiolj77.01.033
- Matselyukh B, Mohammmadipanah F, Laatsch H, Rohr J, Efremenkova O, Khilya V. N-methylphenylalanyl-dehydrobutyrine diketopiperazine, an A-factor mimic that restores antibiotic biosynthesis and morphogenesis in Streptomyces globisporus 1912-B2 and Streptomyces griseus 1439. J. Antibiot. 2015; 68: 9–14. https://doi.org/10.1038/ja.2014.86
- Myronovskyj M, Tokovenko B, Brötz E, Rückert C, Kalinowski J, Luzhetsky A. Genome rearrangement of Streptomyces albus J1074 lead to the carotenoid gene cluster activation. Appl. Microbiol. Biotechnol. 2014; 98: 795–806. https://doi.org/10.1007/s00253-013-5440-6
- Ostash B, Rebets Y, Myronovskyy M, Tsypik O, Kulachkovskyy O, Datsyuk Y, et al. Identification and characterization of the Streptomyces globisporus 1912 regulatory gene lndYR that affects sporulation and antibiotic production. Microbiology. 2011; 157: 1240–1249. https://doi.org/10.1099/mic.0.045088-0
- Romero-Rodriguez A, Robledo-Casados I, Sanchez S. An overview on transcriptional regulators in Streptomyces. Biochim. Biophys. Acta. 2015; 1849: 1017–1039.
- Takano H, Obitsu S, Beppu T, Ueda K. Light-induced carotenogenesis in Streptomyces coelicolor A3(2): identification of an extracytoplasmic function sigma factor that directs photodipendent transcription of the carotenoid biosynthesis gene cluster. J. Bacteriol. 2005; 187: 1825–1832. https://doi.org/10.1128/JB.187.5.1825-1832.2005
- Van Wezel G, McKenzie N, Nodwell J. Applying the genetics of secondary metabolism in model Actinomycetes to the discovery of new antibiotics. Methods Enzymol. 2009; 458: 117–141. https://doi.org/10.1016/S0076-6879(09)04805-8