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两种漆酶的食品级表达及其对霉菌毒素的酶促降解特性研究

Food-Grade Expression of Two Laccases in and Study on Their Enzymatic Degradation Characteristics for Mycotoxins.

作者信息

Sun Zhen, You Yingxin, Xu Huidong, You Yang, He Wenjing, Wang Zhouping, Li Aitao, Xia Yu

机构信息

State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.

School of Life Sciences, Hubei University, Wuhan 430062, China.

出版信息

J Agric Food Chem. 2024 Apr 10. doi: 10.1021/acs.jafc.4c00521.

Abstract

Mycotoxin contamination poses substantial health risks to humans and animals. In this study, the two laccases PpLac1 and AoLac2 from and were selected and heterologously expressed in in a food-grade manner to detoxify aflatoxin B (AFB), zearalenone (ZEN), and deoxynivalenol (DON). Both laccases exhibited degradation activity toward these three mycotoxins, while the efficiency of these for DON was relatively low. Therefore, molecular docking between these laccases and DON was conducted to analyze their potential interaction mechanisms. Furthermore, the degradation conditions of AFB and ZEN by the two laccases were optimized, and the optimal degradation rates for AFB and ZEN by PpLac1 reached 78.51 and 78.90%, while those for AFB and ZEN by AoLac2 reached 72.27 and 80.60%, respectively. The laccases PpLac1 and AoLac2 successfully transformed AFB and ZEN into the compounds AFQ and 15-OH-ZEN, which were 90 and 98% less toxic than the original compounds, respectively. Moreover, the culture supernatants demonstrated effective mycotoxin degradation results for AFB and ZEN in contaminated feed samples. The residual levels of AFB and ZEN in all samples ranged from 6.61 to 8.72 μg/kg and 3.44 to 98.15 μg/kg, respectively, and these levels were below the limit set by the European Union standards. All of the results in this study indicated that the two laccases have excellent application potential in the feed industry.

摘要

霉菌毒素污染对人类和动物构成重大健康风险。在本研究中,从[具体来源1]和[具体来源2]中选择了两种漆酶PpLac1和AoLac2,并以食品级方式在[表达宿主]中进行异源表达,以解毒黄曲霉毒素B(AFB)、玉米赤霉烯酮(ZEN)和脱氧雪腐镰刀菌烯醇(DON)。两种漆酶对这三种霉菌毒素均表现出降解活性,而它们对DON的降解效率相对较低。因此,对这些漆酶与DON进行分子对接,以分析它们潜在的相互作用机制。此外,对两种漆酶降解AFB和ZEN的条件进行了优化,PpLac1对AFB和ZEN的最佳降解率分别达到78.51%和78.90%,而AoLac2对AFB和ZEN的最佳降解率分别达到72.27%和80.60%。漆酶PpLac1和AoLac2成功地将AFB和ZEN转化为AFQ和15-OH-ZEN化合物,其毒性分别比原始化合物低90%和98%。此外,培养上清液对受污染饲料样品中的AFB和ZEN显示出有效的霉菌毒素降解效果。所有样品中AFB和ZEN的残留水平分别为6.61至8.72μg/kg和3.44至98.15μg/kg,这些水平均低于欧盟标准设定的限值。本研究的所有结果表明,这两种漆酶在饲料工业中具有优异的应用潜力。

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