Tracing toxic fumonisins - Fumonisin sensing by Aspergillus niger Fumonisin Amine Oxidase (AnFAO) and amperometric hydrogen peroxide detection
- Fumonisins are a class of toxic secondary metabolites produced by various Fusarium species. The two most important producers of fumonisins are F. verticillioides and F. proliferatum but also Aspergillus niger is known to produce fumonisins. Most frequently they occur on maize, but also other grains can be contaminated with this group of mycotoxins. Exposure to fumonisins by dietary intake can have serious health effects on farm animals such as equine leukoencephalomalacia and porcine pulmonary oedema and is associated with neural tube defects and esophageal cancer in humans. Thus, the European Commission sets legal limits for fumonisins in foodstuffs. The detection of fumonisins is frequently performed in laboratories by chromatographic methods, which are costly and require trained personnel. Simplifying the analysis is therefore a major goal using portable detection systems. Electrochemical enzymatic biosensors offer great promise to meet this demand. Here we report for the first timeFumonisins are a class of toxic secondary metabolites produced by various Fusarium species. The two most important producers of fumonisins are F. verticillioides and F. proliferatum but also Aspergillus niger is known to produce fumonisins. Most frequently they occur on maize, but also other grains can be contaminated with this group of mycotoxins. Exposure to fumonisins by dietary intake can have serious health effects on farm animals such as equine leukoencephalomalacia and porcine pulmonary oedema and is associated with neural tube defects and esophageal cancer in humans. Thus, the European Commission sets legal limits for fumonisins in foodstuffs. The detection of fumonisins is frequently performed in laboratories by chromatographic methods, which are costly and require trained personnel. Simplifying the analysis is therefore a major goal using portable detection systems. Electrochemical enzymatic biosensors offer great promise to meet this demand. Here we report for the first time an enzymatic fumonisin sensing approach with amperometric detection. For this purpose, an Aspergillus niger fumonisin amine oxidase (AnFAO) catalyzing the oxidative deamination of fumonisins, producing hydrogen peroxide, was recombinantly produced in E. coli. For the first time, the specific enzyme activity of AnFAO was determined using a horseradish peroxidase-based fluorescence assay. It was found that the specific activity of AnFAO using 20 μM Fumonisin B1 as substrate is higher than for 20 μM Fumonisin B2 with 0.122 U mg-1 and 0.058 U mg-1, respectively. It was possible to show a dependence of enzyme activity with enzyme – and substrate-concentration. For fumonisin B1 detection, the enzyme was coupled covalently to magnetic particles and the enzymatically produced H2O2 was detected amperometrically in a flow injection system using Prussian blue carbon electrodes. The developed method allows to quantify fumonisin B1 concentrations down to 1.5 µM and demonstrates that the recombinantly produced AnFAO was able to deaminate different concentrations of fumonisin even in immobilized form. Thus, this enzyme is well suited to develop an enzyme based electrochemical biosensor for fumonisin contaminated food and feed.…
Autor*innen: | Soraya RiedelORCiD |
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Koautor*innen: | Rabia Bayram, Sarah Döring, Rudolf SchneiderORCiD, Zoltán KonthurORCiD |
Dokumenttyp: | Posterpräsentation |
Veröffentlichungsform: | Präsentation |
Sprache: | Englisch |
Jahr der Erstveröffentlichung: | 2022 |
Organisationseinheit der BAM: | 1 Analytische Chemie; Referenzmaterialien |
1 Analytische Chemie; Referenzmaterialien / 1.8 Umweltanalytik | |
DDC-Klassifikation: | Naturwissenschaften und Mathematik / Chemie / Analytische Chemie |
Freie Schlagwörter: | Amperometry; Biosensor; Electrochemical sensor; Mycotoxins; Recombinant protein expression |
Themenfelder/Aktivitätsfelder der BAM: | Umwelt |
Umwelt / Sensorik | |
Veranstaltung: | Posterschau Adlershofer Forschungsforum |
Veranstaltungsort: | Berlin, Germany |
Beginndatum der Veranstaltung: | 11.11.2022 |
Bemerkung: | Geburtsname von Riedel, Soraya: Höfs, S. - Birth name of Riedel, Soraya: Höfs, S. |
Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
Datum der Freischaltung: | 15.11.2022 |
Referierte Publikation: | Nein |