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利用胆碱酯酶和固定化发光光细菌进行真菌毒素的快速分析,并评估其酶解效率。

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

机构信息

Faculty of Chemistry, Lomonosov Moscow State University, Lenin Hills 1/3, 119991 Moscow, Russia.

N.M. Emanuel Institute of Biochemical Physics RAS, Kosigina str., 4, 119334 Moscow, Russia.

出版信息

Toxins (Basel). 2021 Jan 6;13(1):34. doi: 10.3390/toxins13010034.

Abstract

Novel sensitive analytical agents that can be used for simple, affordable, and rapid analysis of mycotoxins are urgently needed in scientific practice, especially for the screening of perspective bio-destructors of the toxic contaminants. We compared the characteristics of a rapid quantitative analysis of different mycotoxins (deoxynivalenol, ochratoxin A, patulin, sterigmatocystin, and zearalenone) using acetyl-, butyrylcholinesterases and photobacterial strains of luminescent cells in the current study. The best bioindicators in terms of sensitivity and working range (μg/mL) were determined as follows: sp. 17 cells for analysis of deoxynivalenol (0.8-89) and patulin (0.2-32); sp. 9.2 cells for analysis of ochratoxin A (0.4-72) and zearalenone (0.2-32); acetylcholinesterase for analysis of sterigmatocystin (0.12-219). The cells were found to be more sensitive than enzymes. The assayed strains of photobacterial cells ensured 44%-83% lower limit of detection for deoxynivalenol and sterigmatocystin as compared to the previously known data for immobilized luminescent cells, and the range of working concentrations was extended by a factor of 1.5-3.5. Calibration curves for the quantitative determination of patulin using immobilized photobacteria were presented in this work for the first time. This calibration was applied to estimate the enzyme efficiency for hydrolyzing mycotoxins using zearalenone and His-tagged organophosphorus hydrolase as examples.

摘要

在科学实践中,迫切需要新型的、灵敏的分析试剂,用于简单、经济且快速地分析真菌毒素,特别是用于筛选潜在的有毒污染物的生物解毒剂。在本研究中,我们比较了使用乙酰胆碱酯酶和光细菌发光细胞测定不同真菌毒素(脱氧雪腐镰刀菌烯醇、黄曲霉毒素 A、棒曲霉素、桔青霉素和玉米赤霉烯酮)的快速定量分析的特点。根据灵敏度和工作范围(μg/mL)确定了最佳的生物指示剂如下: sp. 17 细胞用于分析脱氧雪腐镰刀菌烯醇(0.8-89)和棒曲霉素(0.2-32); sp. 9.2 细胞用于分析黄曲霉毒素 A(0.4-72)和玉米赤霉烯酮(0.2-32);乙酰胆碱酯酶用于分析桔青霉素(0.12-219)。与酶相比,细胞更为敏感。与以前已知的固定化发光细胞数据相比,所测定的光细菌菌株确保了脱氧雪腐镰刀菌烯醇和桔青霉素的检测下限降低了 44%-83%,工作浓度范围扩大了 1.5-3.5 倍。本文首次提出了使用固定化光细菌定量测定棒曲霉素的校准曲线。该校准曲线用于以玉米赤霉烯酮和 His 标记的有机磷水解酶为例估计酶对真菌毒素的水解效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2613/7825120/924df49dcf9a/toxins-13-00034-g001.jpg

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