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Selected Publication:

Vekiru, E; Fruhauf, S; Hametner, C; Schatzmayr, G; Krska, R; Moll, WD; Schuhmacher, R.
(2016): Isolation and characterisation of enzymatic zearalenone hydrolysis reaction products
WORLD MYCOTOXIN J. 2016; 9(3): 353-363. FullText FullText_BOKU

Abstract:
Zearalenone (ZEA) is an oestrogenic mycotoxin produced by several Fusarium species, and it frequently contaminates cereals used for food or animal feed. A ZEA-lactonase of Gliocladium roseum was previously described to hydrolyse ZEA to an unstable intermediate, which spontaneously decarboxylates to non-oestrogenic, decarboxylated hydrolysed ZEA (DHZEN). We expressed a codon-optimised version of the ZEA-lactonase (ZHD101) gene of G. roseum MA 918 with a secretion leader in Pichia pastoris and purified the recombinant enzyme from culture supernatant by His-tag mediated affinity chromatography. After incubation of the enzyme with ZEA, we detected the previously elusive primary reaction product hydrolysed ZEA (HZEN) by liquid chromatography tandem mass spectrometry, purified it by preparative high-performance liquid chromatography, and confirmed its postulated structure ((E)-2,4-dihydroxy-6-(10-hydroxy-6-oxo-1-undecen-1-yl) benzoic acid) by nuclear magnetic resonance techniques. Spontaneous decarboxylation to DHZEN ((E)-1-(3,5-dihydroxy-phenyl)-10-hydroxy-1-undecen-6-one), but not to a previously reported isomer, was observed. Biomass resuspensions of G. roseum strains MA 918 and the strains used for previous work, NBRC 7063 and ATCC 8684, all converted ZEA to HZEN, DHZEN, and further unknown metabolites. We studied partitioning of HZEN and DHZEN between aqueous phases and organic solvents, and found that HZEN did not partition into chloroform as extraction solvent, under the conditions used by previous authors. In contrast, extraction with ethyl acetate at pH 2.0 was suitable for simultaneous extraction of HZEN and DHZEN. The detection of HZEN and its availability as an analytical standard may assist further work towards possible application of ZEA-lactonase (e.g. determining kinetic parameters) for detoxification of ZEA.
Authors BOKU Wien:
Krska Rudolf
Kunz-Vekiru Elisavet
Schuhmacher Rainer
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Find related publications in this database (Keywords)
detoxification
lactonase
mass spectrometry
NMR
Gliocladium roseum


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