Microbial metabolomics in open microscale platforms.
Microbial metabolomics in open microscale platforms.
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DOI：
10.1038/ncomms10610
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发表时间：
2016-02-04
影响因子：
16.6
通讯作者：
Berthier E
中科院分区：
文献类型：
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作者：
Barkal LJ;Theberge AB;Guo CJ;Spraker J;Rappert L;Berthier J;Brakke KA;Wang CCC;Beebe DJ;Keller NP;Berthier E
微生物次级代谢组包含巨大的合成多样性，使微生物能够调整其化学反应以适应不断变化的微环境。传统的代谢组学方法不足以探测各种环境或环境动态。在这里，我们介绍一类微型培养平台来分析真菌和细菌次级代谢组的化学多样性。通过利用稳定的双相界面，通过液-液萃取将微培养与小分子分离相结合，我们能够使用质谱进行代谢组学规模的分析。该平台有助于探索培养微环境（包括使用既定方法通常无法访问的稀有培养基）、用于代谢物分离的不寻常有机溶剂和微生物突变体。利用曲霉属（一种以其丰富的次生代谢而闻名的真菌属），我们描述了培养几何形状和生长基质对次生代谢的影响，强调了微尺度系统解锁未知或神秘的次生代谢物以发现天然产物的潜在用途。最后，我们证明了此类微流体系统研究真菌和细菌之间的界间通讯的潜力。 微生物培养和随后的代谢组学的传统方法既耗时又费力。在这里，作者提出了一个具有集成提取功能的微型培养平台，用于相关微环境和微生物共培养的高效、低容量代谢组学。
The microbial secondary metabolome encompasses great synthetic diversity, empowering microbes to tune their chemical responses to changing microenvironments. Traditional metabolomics methods are ill-equipped to probe a wide variety of environments or environmental dynamics. Here we introduce a class of microscale culture platforms to analyse chemical diversity of fungal and bacterial secondary metabolomes. By leveraging stable biphasic interfaces to integrate microculture with small molecule isolation via liquid–liquid extraction, we enable metabolomics-scale analysis using mass spectrometry. This platform facilitates exploration of culture microenvironments (including rare media typically inaccessible using established methods), unusual organic solvents for metabolite isolation and microbial mutants. Utilizing Aspergillus, a fungal genus known for its rich secondary metabolism, we characterize the effects of culture geometry and growth matrix on secondary metabolism, highlighting the potential use of microscale systems to unlock unknown or cryptic secondary metabolites for natural products discovery. Finally, we demonstrate the potential for this class of microfluidic systems to study interkingdom communication between fungi and bacteria. Traditional methods for microbial culture and subsequent metabolomics are time-consuming and labour-intensive. Here the authors present a microscale culture platform with integrated extraction for efficient, low-volume metabolomics of relevant microenvironments and microbial co-cultures.