Evaluation of heterologous promoters for genetic analysis of Actinoplanes teichomyceticus—Producer of teicoplanin, drug of last defense
| Autor: | Flavia Marinelli, Bohdan Ostash, Anton Kobylyanskyy, Nestor Zaburannyi, Olexandr Yushchuk, Victor Fedorenko, Liliya Horbal, Andriy Luzhetskyy |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Overexpression
medicine.drug_class Mutagenesis (molecular biology technique) Bioengineering Glycopeptide antibiotic Actinoplanes teichomyceticus Promoter Reporter gene Teicoplanin Applied Microbiology and Biotechnology Streptomyces Genes Reporter medicine Promoter Regions Genetic Actinoplanes Glucuronidase Regulator gene Genetics biology Micromonosporaceae General Medicine biology.organism_classification Anti-Bacterial Agents Biotechnology medicine.drug |
| Zdroj: | Journal of Biotechnology. 168:367-372 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/j.jbiotec.2013.10.018 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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