Quantifying Metabolic Activity of Filamentous Fungi Using a Colorimetric XTT Assay

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• 作者：Bill J. Moss ; Yonghyun Kim ; M. P. Nandakumar ; Mark R. Marten
• 刊名：Biotechnology Progress
• 出版年：2008
• 出版时间：June 2008
• 年：2008
• 卷：24
• 期：3
• 页码：780 - 783
• 全文大小：52K
• 年卷期：v.24,no.3(June 2008)
• ISSN：1520-6033

文摘

The filamentous nature and robust cell walls of many fungi render traditional measurements ofactive biomass (e.g., turbidity, dry cell weight) ineffective for most fungal bioprocesses. Toovercome this challenge, an assay for quantification of overall metabolic activity is developedusing 2,3-bis(2-methoxy-4-nitro-5-sulfophenly)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT), which in the presence of active mitochondria is converted to a water-solubleformazan derivative that absorbs light in the visible spectrum (430-490 nm). Tests on the modelfungus Aspergillus nidulans show that in actively growing cultures XTT absorbance is linearlyrelated to dry cell weight below 0.2 g/kg broth. Validation through growth rate testing showsthe developed XTT assay is able to accurately quantify reductions in culture metabolism duringdamaging physical treatment (heat, high shear, microwaving). Experiments in batch culturedemonstrate that the developed XTT assay is capable of reporting on metabolic activity wheredry cell weight is not. The developed assay is inexpensive, relatively rapid, and easy to conduct,making it ideally suited for assessment of fungal processes in the biotechnology industry.