Abstract
The mycotoxin patulin is produced by the blue mould pathogen Penicillium expansum in rotting apples during postharvest storage. Patulin is toxic to a wide range of organisms, including humans, animals, fungi and bacteria. Wash water from apple packing and processing houses often harbours patulin and fungal spores, which can contaminate the environment. Ubiquitous epiphytic yeasts, such as Rhodosporidium kratochvilovae strain LS11 which is a biocontrol agent of P. expansum in apples, have the capacity to resist the toxicity of patulin and to biodegrade it. Two non-toxic products are formed. One is desoxypatulinic acid. The aim of the work was to develop rapid, high-throughput bioassays for monitoring patulin degradation in multiple samples. Escherichia coli was highly sensitive to patulin, but insensitive to desoxypatulinic acid. This was utilized to develop a detection test for patulin, replacing time-consuming thin layer chromatography or high-performance liquid chromatography. Two assays for patulin degradation were developed, one in liquid medium and the other in semi-solid medium. Both assays allow the contemporary screening of a large number of samples. The liquid medium assay utilizes 96-well microtiter plates and was optimized for using a minimum of patulin. The semi-solid medium assay has the added advantage of slowing down the biodegradation, which allows the study and isolation of transient degradation products. The two assays are complementary and have several areas of utilization, from screening a bank of microorganisms for biodegradation ability to the study of biodegradation pathways.
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Notes
PBS, Phosphate buffered saline.
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Acknowledgments
We would like to express our sincere thanks to Dr. Anders Falk, who performed the statistical analysis. We are truly grateful to student Paolo D’Apruzzo, who participated in the development of the filter assay. We would like also to acknowledge Dr. Alison Hill, University of Exeter, United Kingdom, for her contributions to the discussion, to Fig. 1 and for the critical reading of the manuscript. We are indebted to Dr. Rosa Durán-Patrón, University of Cádiz, Spain, who enabled C.P.-R. to participate in this work. Dr. Duran-Patrón has been collaborating on the structural elucidation of the biodegradation products of patulin through a bilateral project entitled: “Studio del pathway di biodegradazione della micotossina patulina operata da un lievito basidiomicete”, with funding from the Ministero dell’Istruzione, dell’Università e della Ricerca (project number IT088MB951) in Italy, and by the Ministerio de Ciencia e Innovación (project number HI2007-0026) in Spain. The research was overall funded by the Italian Ministry for University and Scientific Research (MIUR), through the two projects: PRIN n. 2006072204 and “Incentivazione alla mobilità di studiosi stranieri e italiani residenti all’estero”, (DM 1.2.2005, n.18).
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Wright, S.A.I., de Felice, D.V., Ianiri, G. et al. Two rapid assays for screening of patulin biodegradation. Int. J. Environ. Sci. Technol. 11, 1387–1398 (2014). https://doi.org/10.1007/s13762-013-0325-x
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DOI: https://doi.org/10.1007/s13762-013-0325-x