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Six novel constitutive promoters for metabolic engineering of Aspergillus niger

  • Applied genetics and molecular biotechnology
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Abstract

Genetic tools for the fine-tuning of gene expression levels are a prerequisite for rational strain optimization through metabolic engineering. While Aspergillus niger is an industrially important fungus, widely used for production of organic acids and heterologous proteins, the available genetic tool box for this organism is still rather limited. Here, we characterize six novel constitutive promoters of A. niger providing different expression levels. The selection of the promoters was based on published transcription data of A. niger. The promoter strength was determined with the β-glucuronidase (gusA) reporter gene of Escherichia coli. The six promoters covered a GUS activity range of two to three orders of magnitude depending on the strain background. In order to demonstrate the power of the newly characterized promoters for metabolic engineering, they were used for heterologous expression of the cis-aconitate decarboxylase (cad1) gene of Aspergillus terreus, allowing the production of the building block chemical itaconic acid with A. niger. The CAD activity, dependent on the choice of promoter, showed a positive correlation with the specific productivity of itaconic acid. Product titers from the detection limit to up to 570 mg/L proved that the set of constitutive promoters is a powerful tool for the fine-tuning of metabolic pathways for the improvement of industrial production processes.

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Acknowledgments

We acknowledge Karina Schindler for her contribution in plasmid construction and fungal transformation and Stefanie Müller for excellent technical assistance throughout the project. This work has been financially supported by the FHplus Program of the Austrian Research Promotion Agency FFG, Project METORGANIC. Furthermore this work has been supported by the Federal Ministry of Economy, Family and Youth (BMWFJ), the Federal Ministry of Traffic, Innovation and Technology (bmvit), the Styrian Business Promotion Agency SFG, the Standortagentur Tirol, and ZIT—Technology Agency of the City of Vienna through the COMET-Funding Program managed by the Austrian Research Promotion Agency FFG.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. School of Bioengineering, University of Applied Sciences FH-Campus Wien, Muthgasse 56, 1190, Vienna, Austria

    Marzena Blumhoff & Michael Sauer

  2. Department of Biotechnology, BOKU-VIBT University of Natural Resources and Life Sciences, Muthgasse 18, 1190, Vienna, Austria

    Marzena Blumhoff, Hans Marx, Diethard Mattanovich & Michael Sauer

  3. Austrian Centre of Industrial Biotechnology (ACIB GmbH), Muthgasse 11, 1190, Vienna, Austria

    Matthias G. Steiger, Diethard Mattanovich & Michael Sauer

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  1. Marzena Blumhoff
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  2. Matthias G. Steiger
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  3. Hans Marx
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  4. Diethard Mattanovich
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  5. Michael Sauer
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Correspondence to Matthias G. Steiger.

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Blumhoff, M., Steiger, M.G., Marx, H. et al. Six novel constitutive promoters for metabolic engineering of Aspergillus niger . Appl Microbiol Biotechnol 97, 259–267 (2013). https://doi.org/10.1007/s00253-012-4207-9

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  • DOI: https://doi.org/10.1007/s00253-012-4207-9

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