Dynamic metabolomics is a vital tool for industrial biotechnology. Sample preparation approaches dramatically influence data quality and, ultimately, interpretation and conclusions from metabolome experiments. However, standardized protocols for highly reproducible metabolic datasets are limited.
Recently, the research group led by Prof. SUN Jibin and Prof. ZHENG Ping from Tianjin Institute of Industrial Biotechnology of the Chinese Academy of Sciences presented an optimized liquid chromatography-tandem mass spectrometry-based metabolomics methodology for Aspergillus niger, an important industrial workhorse.
It was found that fast filtration with liquid nitrogen was more suitable for cell quenching, due to its minimal disruption to cell integrity, and improved intracellular metabolite recovery.
For intracellular metabolite extraction, acetonitrile/water at -20°C, combined with boiling ethanol extraction protocols, achieved unbiased metabolite profiling.
This improved methodology has been applied to unveil the dynamic metabolite profile of one citrate over-producing A. niger isolate under citrate fermentation.
This study delivers a highly standardized and improved metabolomics methodology and paves the way for systems metabolic engineering of the biotechnologically important fungi.
The study entitled "Comprehensive improvement of sample preparation methodologies facilitates dynamic metabolomics of Aspergillus niger" has been published in Biotechnology Journal.
This study was supported by National Natural Sciences Foundation of China, and Tianjin Science and Technology Project.
Intracellular metabolite profiling of intermediates involved in central metabolic pathway in A. niger during citrate fermentation (Image by TIB)
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