摘要
黄曲霉毒素B_(1)(Aflatoxin B_(1),AFB_(1))是二氢呋喃氧杂萘邻酮的衍生物,是目前已知最强的化学致癌物之一,对人类和动物健康造成严重威胁。研究以铜绿假单胞菌M4的发酵上清液为对象,利用超滤、凝胶过滤色谱和离子交换色谱对AFB_(1)降解酶进行分离纯化,通过超高效液相色谱-电喷雾-离子肼串联质谱仪对纯化的活性酶进行鉴定,得到比活性为2298.49 U/mg的小分子多肽。利用薄层色谱法和超高效液相色谱-飞行时间质谱仪对AFB_(1)降解产物进行解析,结果显示硅胶板上出现了不同于AFB_(1)的新荧光斑点,质谱检测到一个主要降解产物,利用Massynlynx V4.1软件推测降解产物的分子式为C 11 H 10 O 4,并对AFB_(1)降解产物的降解途径进行推导。研究纯化的耐高温小分子多肽可有效将降解AFB_(1),从降解破坏位点推测降解产物可能是低毒或无毒物质,这为进一步开发AFB_(1)脱毒剂提供参考。
Aflatoxin B_(1)(AFB_(1)),a derivative of dihydrofuran coumarin,is one of the strongest known chemical carcinogenic,posing a serious threat to human and animal health.In this study,the degradation enzymes were purified from the supernatant of Pseudomonas aeruginosa M4 fermentation broth by ultrafiltration,gel filtration chromatography,and ion-exchange chromatography.The purified active enzymes were identified by ultra-high-performance liquid chromatography with electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS).The results indicated that the specific activity of the polypeptide was 2298.49 U/mg.The degradation products were analyzed by thin-layer chromatography and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).The results indicated that a new fluorescent spot appeared on the silica plate.A main degradation product was detected by UPLC-Q-TOF-MS.The molecular formula of the degradation product was speculated to be C 11 H 10 O 4 by using Massynlynx V4.1 software.And the degradation pathway of the AFB_(1) degradation product was derived.The high-temperature resistance micromolecule polypeptide purified in this study could effectively degrade AFB_(1),and it may be low or non-toxic substance from the degradation destruction site to infer the degradation product,providing a reference for the further development of AFB_(1) detoxification.
作者
许艳华
董慧燕
娄海伟
王香玉
赵仁勇
Xu Yanhua;Dong Huiyan;Lou Haiwei;Wang Xiangyu;Zhao Renyong(College of Food Science and Engineering,Henan University of Technology,Zhengzhou 450001)
出处
《中国粮油学报》
CSCD
北大核心
2023年第8期33-40,共8页
Journal of the Chinese Cereals and Oils Association
基金
国家自然科学基金-河南联合基金重点项目(U1604234)
河南省现代农业产业技术体系建设专项资金资助项目(S2015-02-06)
河南工业大学粮油食品学科国家工程实验室/省重点实验室开放课题项目(NL2021009)。
作者简介
第一作者:许艳华,女,1991年出生,博士,粮食资源转化与利用,xyhst0906@126.com;通信作者:赵仁勇,男,1969年出生,教授,粮食资源转化与利用,zry8600@haut.edu.cn。
