Activation of a Cryptic Manumycin-Type Biosynthetic Gene Cluster of Saccharothrix espanaensis DSM44229 by Series of Genetic Manipulations

0542621 - MBÚ 2022 RIV CH eng J - Článek v odborném periodiku
Gorniakova, D. - Petříček, M. - Kahoun, D. - Grabic, R. - Zelenka, Tomáš - Chroňáková, Alica - Petříčková, K.
Activation of a Cryptic Manumycin-Type Biosynthetic Gene Cluster of Saccharothrix espanaensis DSM44229 by Series of Genetic Manipulations.
Microorganisms. Roč. 9, č. 3 (2021), č. článku 559. E-ISSN 2076-2607
Grant CEP: GA MZd(CZ) NV17-30091A
Institucionální podpora: RVO:61388971 ; RVO:60077344
Klíčová slova: manumycin * colabomycin * cryptic BGC activation * actinomycetes * Saccharothrix * secondary metabolites * immunomodulators * cancerostatics
Obor OECD: Microbiology; Microbiology (BC-A)
Impakt faktor: 4.926, rok: 2021
Způsob publikování: Open access
https://www.mdpi.com/2076-2607/9/3/559

(1) Background: Manumycins are small actinomycete polyketides with prominent cancerostatic and immunosuppressive activities via inhibition of various eukaryotic enzymes. Their overall activity towards human cells depends on the structural variability of both their polyketide chains, mainly the upper one. In our genetic screening project to find novel producers of anti-inflammatory manumycins, the strain Saccharothrix espanaensis DSM44229 was identified as containing a novel manumycin-type biosynthetic gene cluster (BGC). (2) Methods: The biosynthetic genes appeared to be silent under all assayed laboratory conditions. Several techniques were used to activate the BGC, including: (i) heterologous expression in various hosts, (ii) overexpression of putative pathway-specific regulatory genes, and (iii) overexpression of a bottleneck cyclizing aminolevulinate synthase gene in both natural and heterologous producers. (3) Results: Multiple novel manumycin-type compounds were produced at various levels by genetically-modified strains, sharing a tetraene lower chain structure with a colabomycin subgroup of manumycins, but possessing much shorter and saturated upper chains. (4) Conclusions: A cryptic manumycin-type BGC was successfully activated by genetic means to gain production of novel manumycin-type compounds for future comparative activity assays. Heterologously produced compounds were identical to those found after final activation of the BGC in the original strain, proving the intactness of the cloned BGC.
Trvalý link: http://hdl.handle.net/11104/0320016