Effective Zearalenone Degradation in Model Solutions and Infected Wheat Grain Using a Novel Heterologous Lactonohydrolase Secreted by Recombinant Penicillium canescens

Abstract

This paper reports the first results on obtaining an enzyme preparation that might be promising for the simultaneous decontamination of plant feeds contaminated with a polyketide fusariotoxin, zearalenone (ZEN), and enhancing the availability of their nutritional components. A novel ZEN-specific lactonohydrolase (ZHD) was expressed in a Penicillium canescens strain PCA-10 that was developed previously as a producer of different hydrolytic enzymes for feed biorefinery. The recombinant ZHD secreted by transformed fungal clones into culture liquid was shown to remove the toxin from model solutions, and was able to decontaminate wheat grain artificially infected with a zearalenone-producing Fusarium culmorum. The dynamics of ZEN degradation depending on the temperature and pH of the incubation media was investigated, and the optimal values of these parameters (pH 8.5, 30 °C) for the ZHD-containing enzyme preparation (PR-ZHD) were determined. Under these conditions, the 3 h co-incubation of ZEN and PR-ZHD resulted in a complete removal of the toxin from the model solutions, while the PR-ZHD addition (8 mg/g of dried grain) to flour samples prepared from the infected ZEN-polluted grain (about 16 µg/g) completely decontaminated the samples after an overnight exposure.

Using Cholinesterases and Immobilized Luminescent Photobacteria for the Express-Analysis of Mycotoxins and Estimating the Efficiency of Their Enzymatic Hydrolysis

“…In general, the results obtained are consistent with the literature data on the relatively high hydrolytic activity of lactonases in relation to zearalenone in neutral and slightly alkaline media [ 17 ]; the catalytic activity of the enzyme decreases when the pH is lowered [ 12 , 18 ]. The established regularity, allows us to draw certain conclusions about the prospects of creating food supplements based on His 6 -OPH.…”

“…The controlled release of this enzyme in the digestive system in the areas with alkaline pH values can ensure efficient decomposition of mycotoxins that enter the body of animals with nutrition under the action of His 6 -OPH. His 6 -OPH can be potentially introduced into the feed contaminated with zearalenone, as in the case of recombinant lactonohydrolase [ 12 ], for mycotoxin detoxification. Additionally, using His 6 -OPH in a medium at pH 8.5 provided up to five times faster degradation of zearalenone as compared to recombinant lactonohydrolase expressed in Penicillium canescence .…”

“…These are currently the most actively pursued areas: (i) elucidating the mechanisms of toxic effects of mycotoxins on living organisms [ 5 ]; (ii) development and testing of effective selective and sensitive analytical methods for the detection of mycotoxins in food, agricultural feed, and raw materials for the pharmaceutical industry [ 9 , 10 ]; (iii) search for sorbents capable of removing mycotoxins from raw materials [ 11 ]; (iv) the search for new methods for the destruction of mycotoxins, especially those involving bio-destructors [ 12 , 13 ]. The last of the above directions is especially important today, since it implies the development of means, including combined action, that eliminate mycotoxins not only due to their sorption, but also due to their catalytic decomposition.…”

“…In fundamental and applied research in the field of mycotoxins, liquid chromatography (LC) and enzyme-linked immunosorbent assay (ELISA) are most widely used today. An LC run is followed by mass spectrometry (MS), sensitive fluorescence detection (FLD), or ultraviolet (UV) detection [ 9 , 10 , 12 ]. High analytical accuracy and selectivity are the main advantages of the abovementioned methods.…”

Antifungal Activity of the Dry Biomass of Penicillium chrysogenum F-24-28 and Is Application in Combination with Azoxystrobin for Efficient Crop Protection

“…The majority of these fungi are able to produce highly toxic metabolites (mycotoxins), such as deoxinivalenol, zearalenone, etc., belonging to different groups of chemical compounds. As a rule, fusariotoxins represent chemically stable compounds that are able to be kept unchanged for a long time in plant-derived food and feed [24,25].…”