title: Evaluation of heterologous promoters for genetic analysis of Actinoplanes teichomyceticus--Producer of teicoplanin, drug of last defense.
abstract: Actinoplanes teichomyceticus is the only known producer of the valuable glycopeptide antibiotic teicoplanin. Random mutagenesis and selection were extensively applied to teicoplanin producers, while the gene engineering methods were not used, because of the paucity of genetic tools for A. teichomyceticus. Particularly, availability of promoters of different strength that are functional in Actinoplanes would be very useful for overexpression of beneficial genes. Here we report the use of a glucuronidase reporter system (gusA) for studying transcriptional activity in A. teichomyceticus and describe the behavior of a set of heterologous promoters in this strain. We reveal several elements that exceed in their strength the well-established Streptomyces promoter ermEp, underscoring the utility of the gusA reporter for Actinoplanes sp. Remarkable overproduction of teicoplanin was achieved by constructing strains carrying additional copies of the regulatory gene tcp28 under the control of one of the two most active promoters, moeE5p and actp, discovered in this study.
authors.affiliation: Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 4 Hrushevskoho st, Lviv 79005, Ukraine.
authors.forename: Liliya
authors.initials: L
authors.lastname: Horbal
authors.forename: Anton
authors.initials: A
authors.lastname: Kobylyanskyy
authors.forename: Olexandr
authors.initials: O
authors.lastname: Yushchuk
authors.forename: Nestor
authors.initials: N
authors.lastname: Zaburannyi
authors.forename: Andriy
authors.initials: A
authors.lastname: Luzhetskyy
authors.forename: Bohdan
authors.initials: B
authors.lastname: Ostash
authors.forename: Flavia
authors.initials: F
authors.lastname: Marinelli
authors.forename: Victor
authors.initials: V
authors.lastname: Fedorenko
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000378:metabolism / Q000627:therapeutic use; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D008848:Micromonosporaceae / Q000235:genetics* / Q000254:growth & development; D011401:Promoter Regions, Genetic; D013302:Streptomyces / Q000235:genetics; D017334:Teicoplanin / Q000096:biosynthesis* / Q000378:metabolism / Q000627:therapeutic use
chemical_list: D000900:Anti-Bacterial Agents; D017334:Teicoplanin; D005966:Glucuronidase
keywords: Actinoplanes teichomyceticus; Overexpression; Promoter; Reporter gene; Teicoplanin