Makitrynskyy R. Genes involved in regulation of the biosynthesis of moenomycin A in actinomycetes

Object of the research: mechanisms of regulation of moenomycin biosynthesis in actinomycetes. The aim of the work: to identify the pleiotropic regulatory genes that control moenomycin A biosynthesis, and examine their effect on antibiotic production in S. ghanaensis ATCC14672 and heterologous host carrying moe-gene cluster. Methods: microbiological, molecular genetic, biochemical, mathematical statistics. A system of genetic-engineering manipulations was optimized in S. ghanaensis, namely the introduction of BT1-actinophage based integrative vectors; directed inactivation of genes in the chromosome; utilization of the site-specific recombinases; analysis of transcriptional and translational levels of genes expression using ?-glucuronidase gene uidA as a reporter. By introduction of moeno38-5 cosmid, which carries genes involved in moenomycin biosynthesis, several heterologous hosts for moenomycins production were created. Among them were S. coelicolor, S. lividans, S. albus. Increased concentrations of glucose and nonorganic phosphates in the medium cause considerable reduction of moenomycin production. The influence of several pleiotropic regulatory genes on moenomycin biosynthesis were investigated, namely afsS, afsR, relA, absB, scbA and str6 mutation confers a resistance to streptomycin. AdpAgh, AbsBgh and BldAgh form a complicated and intricate regulatory network that governs moenomycin A biosynthesis and morphological differentiation in S. ghanaensis. Area of use: applied genetics, biotechnology, microbiology.