Aflatoxin-degradative capability of a crude extract from the spent substrate of Pleurotus eryngii cultivation.

Abstract

Biological degradation of mycotoxins is an emerging strategy for detoxification of agricultural commodities. In particular, enzymatic degradation of aflatoxin B1 (AFB1), the most harmful among the mycotoxin known which may occur as contaminant of most of food and feed, has lately raised considerable scientific interest. Ligninolytic enzymes, such as laccase and peroxidise, from white-rot fungi have been proven to be able to break the highly stable molecule of AFB1. However, the high cost of production and purification of these enzymes have limited their implementation into practical technologies aimed at reduction of aflatoxin contamination in the food and feed chains. Among the white-rot fungi there are also cultivable edible mushrooms, such as Pleurotus spp. Every year tons of spent mushroom substrate (SMS) are produced as a by-product of mushroom cultivation and disposed at a cost for farmers. However, SMS may still be a source of bioactive compounds, including ligninolytic enzymes potentially useful for degradation of aflatoxin. We investigated the AFB1-degradative activity of a crude extract (CE) of SMS and undertook a study for characterization of enzyme content and stability of the extracts. CE of SMS was obtained by an extraction buffer (sodium phosphate buffer 0.1 M, pH 7.3) and the extract was incubated with 1 ?g/ml of AFB1 at 25 °C, under continuous shaking at 120 rpm for 1, 3 and 7 days; then the aflatoxin content was determined by ultra performance liquid chromatography (UPLC). After 1 day of incubation, the CE was able to degrade more than 50% of AFB1 and after 3 and 7 days of incubation the percentage of degradation reached the values of 75% and 90%, respectively. The CE contained a high level of laccase activity, quantified in 4 Units per gram of SMS dry weight (U/g DW) and low level of Mn-peroxidase (0.4 U/g DW), as determined by spectrophotometric assays. The enzymatic activity of the CE had its optimum at temperature ranging between 5 and 25 °C and at pH 4, and it was stable at +4 °C for about 60 days. Heat treatment at 100 °C for 10 minutes completely destroyed the degradative activity of CE, and freeze drying resulted in a decrease by 35% of the laccase activity. Based on these preliminary results, SMS proved to be a suitable source of aflatoxin-degrading enzymes and the use of SMS and/or their CE for detoxification of aflatoxin-contaminated commodities, particularly those intended for feed, appears as a coinceivable technology for aflatoxin-free feed production. Further research is needed to improve the stability of SMS extracts and to implement their use in the pipeline of feed processing.


Tutti gli autori

  • Branà M.T.; Sergio L.; Cimmarusti M.T.; Haidukowski M.; Logrieco A.F.; Altomare C.

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2017

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