Heterologous AdpA transcription factors enhance landomycin production in Streptomyces cyanogenus S136 under a broad range of growth conditions
Streptomyces cyanogenus S136 is the only known producer of landomycin A (LaA), one of the largest glycosylated angucycline antibiotics possessing strong antiproliferative properties. There is rising interest in elucidation of mechanisms of action of landomycins, which, in turn, requires access to large quantities of the pure compounds. Overproduction of LaA has been achieved in the past through manipulation of cluster-situated regulatory genes. However, other components of the LaA biosynthetic regulatory network remain unknown. To fill this gap, we elucidated the contribution of AdpA family pleiotropic regulators in landomycin production via expression of adpA genes of different origins in S. cyanogenus S136. Overexpression of the native S. cyanogenus S136 adpA ortholog had no effect on landomycin titers. In the same time, expression of several heterologous adpA genes led to significantly increased landomycin production under different cultivation conditions. Hence, heterologous adpA genes are a useful tool to enhance or activate landomycin production by S. cyanogenus. Our ongoing research effort is focused on identification of mutations that render S. cyanogenus AdpA nonfunctional.
KeywordsStreptomyces Landomycin A Regulation AdpA family regulators
We thank Simon Shaw (Technical University of Denmark, Copenhagen) for careful proofreading of the manuscript.
This work was supported by grants Bg-46F (to V. F.) and M/26-2018 and Bg-41Nr (to B.O.) from the Ministry of Education and Science of Ukraine.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Higashi T, Iwasaki Y, Ohnishi Y, Horinouchi S (2007) A-factor and phosphate depletion signals are transmitted to the grixazone biosynthesis genes via the pathway-specific transcriptional activator GriR. J Bacteriol 189(9):3515–3524. https://doi.org/10.1128/JB.00055-07 CrossRefPubMedPubMedCentralGoogle Scholar
- Higo A, Hara H, Horinouchi S, Ohnishi Y (2012) Genome-wide distribution of AdpA, a global regulator for secondary metabolism and morphological differentiation in Streptomyces, revealed the extent and complexity of the AdpA regulatory network. DNA Res 19(3):259–273. https://doi.org/10.1093/dnares/dss010 CrossRefPubMedPubMedCentralGoogle Scholar
- Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics. John Innes Foundation, NorwichGoogle Scholar
- 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(10):130121. https://doi.org/10.1098/rsob.130121 CrossRefPubMedPubMedCentralGoogle Scholar
- Myronovskyi M, Brötz E, Rosenkränzer B, Manderscheid N, Tokovenko B, Rebets Y, Luzhetskyy A (2016) Generation of new compounds through unbalanced transcription of landomycin A cluster. Appl Microbiol Biotechnol 100(21):9175–9186. https://doi.org/10.1007/s00253-016-7721-3 CrossRefPubMedGoogle Scholar
- Ohnishi Y, Yamazaki H, Kato JY, Tomono A, Horinouchi S (2005) AdpA, a central transcriptional regulator in the A-factor regulatory cascade that leads to morphological development and secondary metabolism in Streptomyces griseus. Biosci Biotechnol Biochem 69(3):431–439. https://doi.org/10.1271/bbb.69.431 CrossRefPubMedGoogle Scholar
- Ostash B, Rix U, Rix LL, Liu T, Lombo F, Luzhetskyy A, Gromyko O, Wang C, Braña AF, Méndez C, Salas JA, Fedorenko V, 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. https://doi.org/10.1016/j.chembiol.2004.03.011 CrossRefPubMedGoogle Scholar
- Panchuk RR, Lehka LV, Terenzi A, Matselyukh BP, Rohr J, Jha AK, Downey T, Kril IJ, Herbacek I, van Schoonhoven S, Heffeter P, Stoika RS, Berger W (2017) Rapid generation of hydrogen peroxide contributes to the complex cell death induction by the angucycline antibiotic landomycin E. Free Radic Biol Med 106:134–147. https://doi.org/10.1016/j.freeradbiomed.2017.02.024 CrossRefPubMedPubMedCentralGoogle Scholar
- Rebets Y, Ostash B, Luzhetskyy A, Hoffmeister D, Brana A, Mendez C, Salas JA, Bechthold A, 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):149–153CrossRefPubMedGoogle Scholar
- Rebets Y, Dutko L, Ostash B, Luzhetskyy A, Kulachkovskyy O, Yamaguchi T, Nakamura T, Bechthold A, 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. https://doi.org/10.1007/s00203-007-0299-5 CrossRefPubMedGoogle Scholar
- Sambrook J, Russell DW (2001) Molecular cloning, a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New-YorkGoogle Scholar
- Xu J, Zhang J, Zhuo J, Li Y, Tian Y, Tan H (2017) Activation and molecular mechanism of a cryptic oviedomycin biosynthetic gene cluster via the disruption of a global regulatory gene adpA in Streptomyces ansochromogenes. J Biol Chem 292(48):19708–19720. https://doi.org/10.1074/jbc.M117.809145 CrossRefPubMedGoogle Scholar
- Yao MD, Ohtsuka J, Nagata K, Miyazono KI, Zhi Y, Ohnishi Y, Tanokura M (2013) Complex structure of the DNA-binding domain of AdpA, the global transcription factor in Streptomyces griseus, and a target duplex DNA reveals the structural basis of its tolerant DNA sequence specificity. J Biol Chem 288:31019–31029. https://doi.org/10.1074/jbc.M113.473611 CrossRefPubMedGoogle Scholar