数字农科院2.0

Fusion strategy enables high-level expression of fungal laccase in Pichia pastoris for efficient detoxification of aflatoxin B1 and zearalenone

文献类型： 外文期刊

作者： Lin Cui;Mengting Liu;Yang He;Ruiju Miao;Huiying Luo;Huoqing Huang;Xiaoyun Su、田健、Tao Tu;Yuan Wang;Xing Qin;Honglian Zhang;Jie Zhang;Yaru Wang;Bin Yao;Yingguo Bai;Xiaolu Wang

作者机构：

关键词： Detoxification;Fusion expression;Laccase;Mycotoxins;Pichia pastoris;Xylanase

期刊名称： Applied Food Research

ISSN： 2772-5022

年卷期： 2025 年 5 卷 2 期

页码：

收录情况： ESCI（2025版）

摘要： Detoxification of aflatoxin B1 (AFB1) and zearalenone (ZEN), which pose substantial threats to both agriculture and food safety, using fungal laccases is an efficient and environmentally friendly technology. However, the difficulty in heterologous expression of fungal laccases has hindered their practical application. In this study, the expression of a Steccherinum ochraceum -originated laccase in Pichia pastoris X33 was achieved through fusion with an Aspergillus niger xylanase ( An XynB). The volumetric activity of laccase was enhanced from an undetectable level to 9,102.0 U/L in a 15-L fermenter. Molecular docking of fusion proteins with mycotoxins indicated that So Lac potentially interacts with AFB1 and ZEN directly. Optimized conditions were established for the mediator-free catalytic transformation of AFB1 and ZEN using the An XynB- So Lac fusion protein, achieving transformation efficiencies of 71.0 % and 72.9 %, respectively, under 50 °C and pH 7.0 for 20 h. The main AFB1 and ZEN transformation products were AFQ1, 13-OH-ZEN-quinone, 15-OH-ZEN, and ZOM-1, respectively. Evaluation of toxicity revealed that the transformed products exhibited markedly reduced toxicity. Furthermore, An XynB- So Lac fusion protein effectively transformed AFB1 and ZEN in raw feed materials, including soybean meal and maize flour. This study describes a novel and feasible strategy for the production of fungal laccases that are difficult to express in P. pastoris in a food-grade manner and illustrated its promise in addressing the detoxification of AFB1 and ZEN residues in food and feed-related matrices.

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