Technical advance in fungal biotechnology: development of a miniaturized culture method and an automated high-throughput screening |
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Authors: | F Alberto D Navarro RP de Vries M Asther E Record |
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Institution: | INRA, Universitéde Provence, Universitéde la Méditerranée, UMR1163, LBCF, Case 932, Marseille, France; Microbiology, Utrecht University, Utrecht, the Netherlands |
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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. |
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Keywords: | Aspergillus niger Aspergillus vadensis automated screening heterologous expression high-throughput method miniaturized cultures |
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