Technical advance in fungal biotechnology: development of a miniaturized culture method and an automated high-throughput screening
Technical advance in fungal biotechnology: development of a miniaturized culture method and an automated high-throughput screening
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DOI：
10.1111/j.1472-765x.2009.02655.x
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发表时间：
2009-08-01
影响因子：
2.4
通讯作者：
Record, E.
中科院分区：
文献类型：
--
作者：
Alberto, F.;Navarro, D.;Record, E.
目的：建立一种可靠、重复性好、快速的培养方法，以筛选大量的真菌转化体。方法和结果：该方法是基于微型细胞培养和微孔板自动表达筛选。为了开发方法，在6天内从黑曲霉中筛选了50个产生阿魏酰酯酶B （FaeB）的重组黑曲霉克隆。然后在烧瓶中检查一组表现出各种行为的克隆，以证明该方法的可重复性。利用该方法，选择了一个产生1个中心点2 g l(-1) FaeB的黑曲霉转化菌株（比黑曲霉过量菌株多12倍）。结论：该微型培养方法可获得可靠、重复性好的结果。与传统的瓶内培养筛选相比，该方法具有高效、节省时间和效率的优点，因为在培养3天后，每天可以筛选800个克隆。研究的意义和影响:这种方法可以应用于其他真菌菌种,酶活性或生物多样性筛查。
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 center dot 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.