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Microcystin-LR Detected in a Low Molecular Weight Fraction from a Crude Extract of Zoanthus sociatus

机译:在低分子量的Zoanthus sociatus粗提取物中检测到微囊藻毒素-LR

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摘要

Cnidarian constitutes a great source of bioactive compounds. However, research involving peptides from organisms belonging to the order Zoanthidea has received very little attention, contrasting to the numerous studies of the order Actiniaria, from which hundreds of toxic peptides and proteins have been reported. In this work, we performed a mass spectrometry analysis of a low molecular weight (LMW) fraction previously reported as lethal to mice. The low molecular weight (LMW) fraction was obtained by gel filtration of a Zoanthus sociatus (order Zoanthidea) crude extract with a Sephadex G-50, and then analyzed by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry (MS) in positive ion reflector mode from m/z 700 to m/z 4000. Afterwards, some of the most intense and representative MS ions were fragmented by MS/MS with no significant results obtained by Protein Pilot protein identification software and the Mascot algorithm search. However, microcystin masses were detected by mass-matching against libraries of non-ribosomal peptide database (NORINE). Subsequent reversed-phase C18 HPLC (in isocratic elution mode) and mass spectrometry analyses corroborated the presence of the cyanotoxin Microcystin-LR (MC-LR). To the best of our knowledge, this finding constitutes the first report of MC-LR in Z. sociatus, and one of the few evidences of such cyanotoxin in cnidarians.
机译:刺鼻虫是生物活性化合物的重要来源。但是,与来自棘轮蛾属的生物体肽有关的研究很少受到关注,这与猕猴桃属的众多研究相反,据报道,已有数百种毒性肽和蛋白质被报道。在这项工作中,我们对以前据称对小鼠具有致命性的低分子量(LMW)馏分进行了质谱分析。低分子量(LMW)馏分是通过用Sephadex G-50凝胶过滤Zoanthus sociatus(Zoanthidea订单)粗提取物获得的,然后通过基质辅助激光解吸/电离飞行时间/飞行时间进行分析。在正离子反射器模式下从m / z 700到m / z 4000进行飞行(MALDI-TOF / TOF)质谱(MS)。随后,一些强度最高且最具代表性的MS离子被MS / MS碎片化,没有明显的通过Protein Pilot蛋白质识别软件和Mascot算法搜索获得的结果。但是,通过与非核糖体肽数据库(NORINE)的库进行质量匹配来检测微囊藻毒素的质量。随后的反相C18 HPLC(等度洗脱模式)和质谱分析证实了氰毒素微囊藻毒素-LR(MC-LR)的存在。据我们所知,这一发现构成了在Z. sociatus中首次出现MC-LR的报道,并且是在刺胞动物中这种氰毒素的少数证据之一。

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