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Genetic manipulation of pathway regulation for overproduction of angucycline-like antibiotic auricin in Streptomyces aureofaciens CCM 3239

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Abstract

The polyketide gene cluster aur1 is responsible for the production of the antibiotic auricin in Streptomyces aureofaciens CCM 3239. Auricin production is low and strictly regulated by two regulators, Aur1P and Aur1R. To improve auricin yield, we genetically manipulated S. aureofaciens CCM 3239 strain to overcome this strict regulation. A regulatory region including aur1R, aur1P, aur1O and the target biosynthetic aur1Ap promoter were replaced by the strong constitutive ermEp* promoter. However, auricin production was decreased in such a genetically manipulated strain. In the second strategy we placed the aur1P gene for auricin pathway-specific activator under the control of the ermEp* promoter. The resulting strain has been shown to produce 2.8-fold higher amount of auricin compared with the WT strain.

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Abbreviations

Am:

Apramycin

CFU:

Colony-forming units

DIG:

Digoxigenin

Km:

Kanamycin

TSP:

Transcription start point

Ap:

Ampicilline

cpm:

Counts per minute

HPLC:

High-pressure liquid chromatography

TLC:

Thin-layer chromatography

WT:

Wild type

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Acknowledgements

We are grateful to Dr. Mark Buttner for the plasmid pPM929, to Prof. Andreas Bechthold for the plasmid pUWLoriT, and to Dr. Bertold Gust for kindly providing all the plasmids and strains used in PCR-targeting system; the system has been supplied by Plant Bioscience Ltd. (Norwich, UK). We thank Ludovit Skultety for his assistance with the mass spectrometric analysis. This work was supported by the Slovak Research and Development Agency under the contract No. APVV-0017-07 the VEGA grant 2/0104/09 from Slovak Academy of Sciences. This publication is also the result of the project implementation: ‘TRANSMED, ITMS: 26240120008’, supported by the Research & Development Operational Programme funded by the ERDF.

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Authors and Affiliations

  1. Institute of Molecular Biology, Slovak Academy of Sciences, 845 51, Bratislava, Slovak Republic

    Renata Novakova, Alena Rehakova, Lubomira Feckova, Peter Kutas, Renata Knischova & Jan Kormanec

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  1. Renata Novakova
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  2. Alena Rehakova
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  3. Lubomira Feckova
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  4. Peter Kutas
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  5. Renata Knischova
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  6. Jan Kormanec
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Correspondence to Jan Kormanec.

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Novakova, R., Rehakova, A., Feckova, L. et al. Genetic manipulation of pathway regulation for overproduction of angucycline-like antibiotic auricin in Streptomyces aureofaciens CCM 3239. Folia Microbiol 56, 276–282 (2011). https://doi.org/10.1007/s12223-011-0033-1

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  • DOI: https://doi.org/10.1007/s12223-011-0033-1

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