Recombinant Expression of Trametes versicolor Aflatoxin B1-Degrading Enzyme (TV-AFB1D) in Engineering Pichia pastoris GS115 and Application in AFB1 Degradation in AFB1-Contaminated Peanuts

Abstract

Aflatoxins seriously threaten the health of humans and animals due to their potential carcinogenic properties. Enzymatic degradation approach is an effective and environmentally friendly alternative that involves changing the structure of aflatoxins. In this study, Trametes versicolor aflatoxin B1-degrading enzyme gene (TV-AFB1D) was integrated into the genome of Pichia pastoris GS115 by homologous recombination approach. The recombinant TV-AFB1D was expressed in engineering P. pastoris with a size of approximately 77 kDa under the induction of methanol. The maximum activity of TV-AFB1D reached 17.5 U/mL after the induction of 0.8% ethanol (v/v) for 84 h at 28 °C. The AFB1 proportion of 75.9% was degraded using AFB1 standard sample after catalysis for 12 h. In addition, the AFB1 proportion was 48.5% using AFB1-contaminated peanuts after the catalysis for 18 h at 34 °C. The recombinant TV-AFB1D would have good practical application value in AFB1 degradation in food crops. This study provides an alternative degrading enzyme for the degradation of AFB1 in aflatoxin-contaminated grain and feed via enzymatic degradation approach.

Advanced mycotoxin control and decontamination techniques in view of an increased aflatoxin risk in Europe due to climate change

“…A wide variety of enzymes have been identified and characterized for their mycotoxin degrading capabilities in the last years (Loi et al, 2017;Wang et al, 2022) (Loi et al, 2016(Loi et al, , 2020aYang et al, 2021), currently there are no commercial options onto the market. Most of the literature focuses on in vitro AFs degradation, lacks data on degradation products, and toxicity assessments.…”

A NEW PRODUCER OF A RECOMBINANT AFLATOXIN-DEGRADING ENZYME OBTAINED VIA HETEROLOGOUS EXPRESSION IN Pichia pastoris

“…Представленные нами данные свидетельствуют о достаточно высоком биотехнологическом потенциале нового продуцента рекомбинантного ADTZ и расширяют пока что ограниченный спектр полученных с помощью системы гетерологичной экспрессии в P. pastoris рекомбинантных ферментов других ксилоторофных грибов, разлагающих АФВ1 (39).…”

Microbial Enzymes Involved in the Biotransformation of Major Mycotoxins

“…Yang et al expressed TV-AFB1D isolated from Trametes versicolor in P. pastoris , which could degrade 49% of AFB1 in peanut samples within 12 h. After induction with 0.8% ethanol (v/v) for 84 h at 28 °C, the maximum activity of TV-AFB 1 D was 17.5 U/mL. In another study, Yang et al employed molecular docking and site-directed mutagenesis to enhance the effectiveness of TV-AFB 1 D on AFB 1 .…”

“…The two AFB 1 degrading enzymes, namely, Lac010 and Chia010, were discovered by sequencing the whole BL010 genome using a bioinformatics technique, and the degradation product of the reaction was C 12 H 14 O 4 determined by quadrupole time-of- flight liquid chromatography−mass spectrometry (LC−Q-TOF/MS). 61 Yang et al 62 expressed TV-AFB1D isolated from Trametes versicolor in P. pastoris, which could degrade 49% of AFB1 in peanut samples within 12 h. After induction with 0.8% ethanol (v/v) for 84 h at 28 °C, the maximum activity of TV-AFB 1 D was 17.5 U/mL. In another study, Yang et al 63 employed molecular docking and site-directed mutagenesis to enhance the effectiveness of TV-AFB 1 D on AFB 1 .…”