The pathway-specific regulatory genes, tei15* and tei16*, are the master switches of teicoplanin production in Actinoplanes teichomyceticus
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Pathogenic antibiotic-resistant bacteria are an unprecedented threat to health care worldwide. The range of antibiotics active against these bacteria is narrow; it includes teicoplanin, a “last resort” drug, which is produced by the filamentous actinomycete Actinoplanes teichomyceticus. In this report, we determine the functions of tei15* and tei16*, pathway-specific regulatory genes that code for StrR- and LuxR-type transcriptional factors, respectively. The products of these genes are master switches of teicoplanin biosynthesis, since their inactivation completely abolished antibiotic production. We show that Tei15* positively regulates the transcription of at least 17 genes in the cluster, whereas the targets of Tei16* still remain unknown. Integration of tei15* or tei16* under the control of the aminoglycoside resistance gene aac(3)IV promoter into attBϕC31 site of the A. teichomyceticus chromosome increased teicoplanin productivity to nearly 1 g/L in TM1 industrial medium. The expression of these genes from the moderate copy number episomal vector pKC1139 led to 3–4 g/L teicoplanin, while under the same conditions, wild type produced approximately 100 mg/L. This shows that a significant increase in teicoplanin production can be achieved by a single step of genetic manipulation of the wild-type strain by increasing the expression of the tei regulatory genes. This confirms that natural product yields can be increased using rational engineering once suitable genetic tools have been developed. We propose that this new technology for teicoplanin overproduction might now be transferred to industrial mutants of A. teichomyceticus.
KeywordsActinoplanes Teicoplanin Pathway-specific regulators Strain improvement Overproducer
This work was supported by grant Bg-98 F of Ministry of education and science of Ukraine (to VF) and by the DAAD Researh Fellowship to LH (PKZ A/13/03150), grant from Fondo di Ateneo per la Ricerca to FM, by Federation of European Microbiological Societies (FEMS) Research Fellowship to AK. Authors also thank the support from Consorzio Interuniversitario per le Biotecnologie (CIB). Authors are particularly grateful to Luciano Gastaldo for his assistance in fermentations.
Conflict of interest
The authors declare that they have no conflicts of interest.
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