Technical advance in fungal biotechnology: development of a miniaturized culture method and an automated high-throughput screening

被引:23
作者
Alberto, F. [1 ]
Navarro, D. [1 ]
de Vries, R. P. [2 ]
Asther, M. [1 ]
Record, E. [1 ]
机构
[1] Univ Aix Marseille 1, Univ Mediterranee, INRA, LBCF,UMR1163, F-13288 Marseille 09, France
[2] Univ Utrecht, Utrecht, Netherlands
关键词
Aspergillus niger; Aspergillus vadensis; automated screening; heterologous expression; high-throughput method; miniaturized cultures; ASPERGILLUS-NIGER; FERULOYL ESTERASE; PROTEIN EXPRESSION; RECOMBINANT PROTEINS; FILAMENTOUS FUNGI; ESCHERICHIA-COLI; PELLET FORMATION; RHIZOPUS-ORYZAE; GENE; STRATEGIES;
D O I
10.1111/j.1472-765X.2009.02655.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:278 / 282
页数:5
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