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Technical advance in fungal biotechnology: development of a miniaturized culture method and an automated high-throughput screening
Authors:F Alberto  D Navarro  RP de Vries  M Asther  E Record
Institution: INRA, Universitéde Provence, Universitéde la Méditerranée, UMR1163, LBCF, Case 932, Marseille, France;
 Microbiology, Utrecht University, Utrecht, the Netherlands
Abstract:Aims:  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.
Methods and Results:  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).
Conclusions:  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.
Significance and Impact of the Study:  This method could be applied to any other fungal strain culture, enzyme activity or biodiversity screening.
Keywords:Aspergillus niger                        Aspergillus vadensis            automated screening  heterologous expression  high-throughput method  miniaturized cultures
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