Rapid Contamination Detection in Validamycin A Production by HS-SPME/GC-MS

Xiao-Mei Yu, Jing Jiang, Shangtian Yang, Wen-Wen Zhou
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Abstract

Validamycin A (Val-A) is one of the most widely used agricultural antibiotics in East Asia especially for controlling rice sheath blight disease. Fermentation contamination of the industrial Val-A producing strain is a common occurrence. Fermentation culture of S. hygroscopicus 5008 has a special smell that could be distinguished from other tainted samples. The change of the volatiles in untainted and tainted samples was characterized using headspace solid phase microextraction (HS-SPME) combined with gas chromatography- mass spectrometry (GC-MS). Seventy-one volatile compounds (including alkanes, amines, alcohols, esters, aldehydes and others) were identified and there were significant differences in the composition of volatiles among different samples. Principal component analysis (PCA) based on the GC-MS data was used to identify the important volatile compounds that contributed to the differentiation of the fermentation samples under different fermentation stages, as well as among different pollution species and fermentation media. Contamination could be discovered in time irrespective of the stage of fermentation and the contaminating bacteria in broth. It is the first report to detect contamination by volatile compounds in the antibiotic fermentation and it was proved that HS-SPME/GC-MS is an effective contamination detection method in Val-A production.
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高效液相色谱-质谱联用技术快速检测缬霉素A生产过程中的污染
缬霉素A (valamycin A, Val-A)是东亚地区应用最广泛的农业抗生素之一,尤其用于防治水稻纹枯病。发酵污染的工业vala生产菌株是一个常见的事件。吸湿菌5008的发酵培养物有一种特殊的气味,可以与其他污染样品区分开来。采用顶空固相微萃取法(HS-SPME)结合气相色谱-质谱法(GC-MS)对未污染和污染样品中挥发物的变化进行了表征。共鉴定出71种挥发性化合物(包括烷烃、胺类、醇类、酯类、醛类等),不同样品的挥发性成分存在显著差异。基于GC-MS数据的主成分分析(PCA)鉴定了在不同发酵阶段、不同污染种类和发酵介质中导致发酵样品差异的重要挥发性化合物。无论发酵期和发酵液中的污染菌数如何,都能及时发现污染。本文首次报道了抗生素发酵过程中挥发性污染物的检测方法,证明了HS-SPME/GC-MS是一种有效的抗生素发酵过程中挥发性污染物的检测方法。
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