Research Article: Process development for the elucidation of mycotoxin formation in Alternaria alternata

Date Published: October 4, 2011

Publisher: Springer

Author(s): Katrin Brzonkalik, Tanja Herrling, Christoph Syldatk, Anke Neumann.


The black mould Alternaria alternata produces a wide diversity of mycotoxins which are of particular health concern. Since no maximum allowable limits are set for Alternaria toxins in food and feed, prevention of Alternaria infestations and mycotoxin spoilage is the only way to avoid health risks. Thus, the understanding of mycotoxin biosynthesis is essential. For that purpose, a reliable batch process in a 2 L bioreactor was established which enables the study of several parameters influencing the production of the mycotoxins alternariol (AOH), alternariol monomethylether (AME) and tenuazonic acid (TA) by A. alternata DSM 12633. Modified Czapek-Dox medium was used with glucose as carbon source and ammonium and nitrate as nitrogen sources. Consumption of carbon and nitrogen sources as well as formation of the three mycotoxins were monitored; the average data of five independent fermentations was plotted and fitted using a logistic equation with four parameters. Maximum mycotoxin concentrations of 3.49 ± 0.12 mg/L AOH, 1.62 ± 0.14 mg/L AME and 38.28 ± 0.1 mg/L TA were obtained.

Partial Text

Mycotoxins are secondary metabolites of low molecular weight produced by filamentous fungi. Since the discovery of the first mycotoxins, the aflatoxins, in 1960 which caused the death of 10,000 turkeys many new mycotoxins have been identified in the last 50 years. Today 300 to 400 compounds are designated as mycotoxins (Bennett and Klich 2003). As other secondary metabolites mycotoxins are formed subsequently to the growth phase and are not necessary for growth or development (Fox and Howlett 2008). Mycotoxin formation is subjected to a complex regulation, but it is often induced by nutrient limitation (Demain 1986). Mycotoxins are released by the fungus in the surrounding substrate and contamination of agricultural products is therefore possible. They are connected to certain health disorders and elicit acute toxic, mutagenic, teratogenic, carcinogenic and sometimes estrogenic properties (Bhatnagar et al. 2002). Based on estimations of the Food and Agriculture Organization (FAO) of the United Nations approximately 25% of the world’s food crops are affected by mycotoxin producing fungi and global losses of foodstuffs due to mycotoxins are in the range of 1000 million tons per year

As it was shown with our results Alternaria toxins can be produced reproducibly in a bioreactor system under controlled conditions. Consumption of nutrient and mycotoxin formation can be characterized with logistic equations. The semi-synthetic Czapek-Dox broth is perfectly suitable for the elucidation of nutritional influences as it was shown previously by (Brzonkalik et al. 2011). Therefore, this medium was chosen for the fermentation experiments, but a further enhancement of mycotoxin production can be achieved by using other complex media.

The authors declare that they have no competing interests.




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