Investigating the mechanism of Bacillus amyloliquefaciens YUAD7 degrading aflatoxin B1 in alfalfa silage using isotope tracing and nuclear magnetic resonance methods

IF 5.2 2区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Ying Tang, Xiaojing Liu, Linlin Tang, Jianxun Dong
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引用次数: 0

Abstract

Background

Fungal toxins are highly toxic and widely distributed, presenting a considerable threat to global agricultural development. Addressing the issue of aflatoxin B1 (AFB1) contamination in feed, it is crucial to ascertain the effectiveness and mechanisms of microbial strains in degradation.

Results

This study used isotope tracing and nuclear magnetic resonance (NMR) to investigate the degradation products of Bacillus amyloliquefaciens YUAD7 in complex substrates. By tracing 14C34-AFB1 and utilizing NMR, ultra-performance liquid chromatography–quadrupole time-of-flight/mass spectrometry (UPLC–Q-TOF/MS) purification and identification techniques, it was confirmed that AFB1 was degraded by YUAD7 into C12H14O4, C5H12N2O2, C10H14O2, and C4H12N2O, effectively removing 99.7% of AFB1 (100 μg/kg) from alfalfa silage. YUAD7 targeted the ester bond in the vanillin lactone ring structure, the ether bond in the furan ring structure, and the unsaturated carbon–carbon double bond in the furan ring structure during AFB1 degradation, disrupting the toxic sites responsible for AFB1's carcinogenic, teratogenic, and mutagenic effects and achieving biodegradation. Moreover, B. amyloliquefaciens YUAD7 transformed AFB1 through processes like hydrogenation, enzyme modification, and the loss of the -CO group while also being associated with metabolic pathways such as alanine, aspartate, glutamate metabolism, glutathione metabolism, cysteine and methionine metabolism, and pentose and glucuronate interconversions.

Conclusions

The utilization of isotope tracing allowed for rapid identification of degradation products in complex substrates, while NMR elucidated the structures of these products. This deepens our understanding of AFB1 biodegradation mechanisms, providing technical support for the practical application of these bacteria in degradation, and new insights into studying the biological degradation mechanism. B. amyloliquefaciens YUAD7 can be used as a potential strain for degrading AFB1 in large-scale silage.

Graphical Abstract

利用同位素示踪和核磁共振方法研究淀粉芽孢杆菌 YUAD7 降解苜蓿青贮中黄曲霉毒素 B1 的机理
真菌毒素毒性强、分布广,对全球农业发展构成了巨大威胁。针对饲料中的黄曲霉毒素 B1(AFB1)污染问题,确定微生物菌株的降解效果和机制至关重要。本研究利用同位素示踪和核磁共振(NMR)技术研究了淀粉芽孢杆菌 YUAD7 在复杂底物中的降解产物。通过追踪 14C34-AFB1,并利用核磁共振、超高效液相色谱-四极杆飞行时间/质谱(UPLC-Q-TOF/MS)纯化和鉴定技术,证实 YUAD7 可将 AFB1 降解为 C12H14O4、C5H12N2O2、C10H14O2 和 C4H12N2O,从而有效去除苜蓿青贮中 99.7% 的 AFB1(100 μg/kg)。在降解 AFB1 的过程中,YUAD7 针对香草醛内酯环结构中的酯键、呋喃环结构中的醚键和呋喃环结构中的不饱和碳碳双键,破坏了导致 AFB1 致癌、致畸和致突变作用的毒性位点,实现了生物降解。此外,淀粉芽孢杆菌 YUAD7 通过氢化、酶修饰、失去 -CO 基团等过程转化了 AFB1,同时还与丙氨酸、天门冬氨酸、谷氨酸代谢、谷胱甘肽代谢、半胱氨酸和蛋氨酸代谢、戊糖和葡萄糖醛酸盐相互转化等代谢途径有关。利用同位素追踪技术可快速鉴定复杂底物中的降解产物,而核磁共振则可阐明这些产物的结构。这加深了我们对 AFB1 生物降解机制的理解,为这些细菌在降解领域的实际应用提供了技术支持,也为研究生物降解机制提供了新的视角。淀粉芽孢杆菌 YUAD7 可作为大规模青贮饲料中降解 AFB1 的潜在菌株。
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来源期刊
Chemical and Biological Technologies in Agriculture
Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.80
自引率
3.00%
发文量
83
审稿时长
15 weeks
期刊介绍: Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.
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