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Aspergillus niger is a superior expression host for the production of bioactive fungal cyclodepsipeptides
Boecker, Simon; Grätz, Stefan; Kerwat, Dennis; Adam, Lutz; Schirmer, David; Richter, Lennart; Schütze, Tabea; Petras, Daniel; Süßmuth, Roderich D.; Meyer, Vera
Background: Fungal cyclodepsipeptides (CDPs) are non-ribosomally synthesized peptides produced by a variety of flamentous fungi and are of interest to the pharmaceutical industry due to their anticancer, antimicrobial and anthelmintic bioactivities. However, both chemical synthesis and isolation of CDPs from their natural producers are limited due to high costs and comparatively low yields. These challenges might be overcome by heterologous expression of the respective CDP-synthesizing genes in a suitable fungal host. The well-established industrial fungus Aspergillus niger was recently genetically reprogrammed to overproduce the cyclodepsipeptide enniatin B in g/L scale, suggesting that it can generally serve as a high production strain for natural products such as CDPs. In this study, we thus aimed to determine whether other CDPs such as beauvericin and bassianolide can be produced with high titres in A. niger, and whether the generated expression strains can be used to synthesize new-to-nature CDP derivatives.
Results: The beauvericin and bassianolide synthetases were expressed under control of the tuneable Tet-on promoter, and titres of about 350–600 mg/L for bassianolide and beauvericin were achieved when using optimized feeding conditions, respectively. These are the highest concentrations ever reported for both compounds, whether isolated from natural or heterologous expression systems. We also show that the newly established Tet-on based expression strains can be used to produce new-to-nature beauvericin derivatives by precursor directed biosynthesis, including the compounds 12-hydroxyvalerate-beauvericin and bromo-beauvericin. By feeding deuterated variants of one of the necessary precursors (d-hydroxyisovalerate), we were able to purify deuterated analogues of beauvericin and bassianolide from the respective A. niger expression strains. These deuterated compounds could potentially be used as internal standards in stable isotope dilution analyses to evaluate and quantify fungal spoilage of food and feed products.
Conclusion: In this study, we show that the product portfolio of A. niger can be expanded from enniatin to other CDPs such as beauvericin and bassianolide, as well as derivatives thereof. This illustrates the capability of A. niger to produce a range of diferent peptide natural products in titres high enough to become industrially relevant.
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- The Correction to this article has been published in Fungal Biology and Biotechnology 2018 5:7. https://fungalbiolbiotech.biomedcentral.com/articles/10.1186/s40694-018-0051-8.
- Eine Korrektur zu diesem Artikel wurde publiziert in Fungal Biology and Biotechnology 2018 5:7. https://fungalbiolbiotech.biomedcentral.com/articles/10.1186/s40694-018-0051-8.
