Technical advance in fungal biotechnology: development of a miniaturized culture method and an automated high-throughput screening
2009
Alberto, F. | Navarro, D. | de Vries, R.P. | Asther, M. | Record, E.
The goal of the study was to develop a reliable, reproducible and rapid method of culture in order to screen a large number of fungal transformants. The method is based upon miniaturized cell cultures and automated expression screening in microwell plates. For the method development, 50 recombinant Aspergillus vadensis clones producing feruloyl esterase B (FaeB) from Aspergillus niger were screened in 6 days. Then a panel of clones showing various behaviours was checked in flasks in order to demonstrate the reproducibility of the method. Using this method, a transformant of A. vadensis producing 1·2 g l⁻¹ of FaeB was selected (12-fold more than the A. niger overproducing strain). This miniaturized culture method allows to obtain reliable and reproducible results. The procedure has the advantages of being efficient, time-saving and more efficient than conventional in-flask culture screening as it can screen 800 clones per day after a culture of 3 days. This method could be applied to any other fungal strain culture, enzyme activity or biodiversity screening.
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