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Metabolomics of the bio-degradation process of aflatoxin B1 by actinomycetes at an initial pH of 6.0

机译:黄曲霉毒素B1在初始pH值为6.0时由放线菌生物降解过程的代谢组学

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摘要

Contamination of food and feed by Aflatoxin B1 (AFB1) is a cause of serious economic and health problems. Different processes have been used to degrade AFB1. In this study, biological degradation of AFB1 was carried out using three Actinomycete species, Rhodococcus erythropolis ATCC 4277, Streptomyces lividans TK 24, and S. aureofaciens ATCC 10762, in liquid cultures. Biodegradation of AFB1 was optimised under a range of temperatures from 25 to 40 °C and pH values of 4.0 to 8.0. An initial concentration of 20 µg/mL of AFB1 was used in this study. The amount of AFB1 remaining was measured against time by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC), coupled with UV and mass spectrometry (LC-MS). All species were able to degrade the AFB1, and no significant difference was found between them. AFB1 remained in the liquid culture for R. erythropolis, S. lividans and S. aureofaciens were 0.81 µg/mL, 2.41 µg/mL and 2.78 µg/mL respectively, at the end of the first 24 h. Degradation occurred at all incubation temperatures and the pH with the optimal conditions for R. erythropolis was achieved at 30 °C and pH 6, whereas for S. lividans and S. aureofaciens the optimum conditions for degradation were 30 °C and pH 5. Analysis of the degradative route indicated that each microorganism has a different way of degrading AFB1. The metabolites produced by R. erythropolis were significantly different from the other two microorganisms. Products of degradation were identified through metabolomic studies by utilizing high-resolution mass spectral data. Mass spectrometric analysis indicated that the degradation of AFB1 was associated with the appearance of a range of lower molecular weight compounds. The pathway of degradation or chemical alteration of AFB1 was followed by means of high resolution Fourier transform mass spectrometry (HR-FTMS) analysis as well as through the MS2 fragmentation to unravel the degradative pathway for AFB1. AFB1 bio-degradation was coupled with the accumulation of intermediates of fatty acid metabolism and glycolysis. A plausible mechanism of degradation of AFB1 by Rhodococcus was hypothesized.
机译:黄曲霉毒素B1(AFB1)对食品和饲料的污染是造成严重经济和健康问题的原因。已经使用了不同的过程来降级AFB1。在这项研究中,在液体培养中,使用三种放线菌属物种红球菌红球菌ATCC 4277,淡链霉菌TK 24和金黄色葡萄球菌ATCC 10762对AFB1进行了生物降解。在温度25到40°C和pH值4.0到8.0的范围内优化了AFB1的生物降解。在这项研究中,AFB1的初始浓度为20 µg / mL。通过薄层色谱(TLC)和高效液相色谱(HPLC)结合紫外和质谱(LC-MS)随时间测量剩余的AFB1量。所有物种都能够降解AFB1,并且它们之间没有发现显着差异。在最初的24 h结束时,残留在红细菌中的AFB1分别为0.81 µg / mL,葡萄球菌和2.41 µg / mL,而金黄色葡萄球菌为2.78 µg / mL。在所有温育温度下都会发生降解,并且在30°C和pH 6时,红球菌的最佳条件pH达到了,而对于S. lividans和金黄色葡萄球菌,最佳降解条件是30°C和pH5。分析降解途径的结果表明每种微生物具有不同的降解AFB1的方式。红血球菌产生的代谢产物与其他两种微生物明显不同。通过代谢组学研究,利用高分辨率质谱数据鉴定了降解产物。质谱分析表明,AFB1的降解与一系列低分子量化合物的出现有关。通过高分辨率傅里叶变换质谱(HR-FTMS)分析以及通过MS2片段化来阐明AFB1的降解或化学变化途径,以阐明AFB1的降解途径。 AFB1的生物降解与脂肪酸代谢和糖酵解中间体的积累有关。推测了红球菌降解AFB1的合理机制。

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