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Influence of pH and carbon to nitrogen ratio on mycotoxin production by Alternaria alternata in submerged cultivation
Production of the Alternaria mycotoxins alternariol (AOH), alternariol monomethylether (AME) and tenuazonic acid (TA) by Alternaria alternata DSM 12633 was influenced by pH and carbon to nitrogen (C:N) ratio of the growth medium both in shaking flasks and bioreactor cultivation. The impact of medium pH on mycotoxin production was studied in the range of pH 3.5 - 8. pH values above 5.5 led to a decreased mycotoxin production or inhibited mycotoxin formation completely whereas an acidic pH in the range of 4.0-4.5 was optimal for mycotoxin production. The influence of the C:N ratio was evaluated over the range of 24 to 96. Glucose was used as carbon source and its concentration was altered while nitrogen concentration was kept constant. Growth kinetics and mycotoxin production parameters were studied depending on different C:N ratios. With increasing initial glucose concentration fungal biomass did increase but the maximum specific growth rate was not influenced. The optimal initial C:N ratio for attaining highest mycotoxin concentrations was 72. A higher C:N ratio did not further enhance mycotoxin production.
Alternaria alternata; Mycotoxins; pH value; C:N ratio; Submerged cultivation
Process development for the elucidation of mycotoxin formation in Alternaria alternata
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.
In this system the effect of different aeration rates (0.53 vvm-0.013 vvm) was tested which exerted a great influence on mycotoxin production. The use of the semi-synthetic Czapek-Dox medium allowed the exchange of carbon and nitrogen sources for acetate and aspartic acid. The use of acetate instead of glucose resulted in the sole production of alternariol whereas the exchange of ammonium and nitrate for aspartate enhanced the production of both AOH and AME while TA production was not affected.
Alternaria alternata; Mycotoxin; Batch process; Aeration rate
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