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

Lett Appl Microbiol. 2009 Aug;49(2):278-82. doi: 10.1111/j.1472-765X.2009.02655.x. Epub 2009 May 27.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus / enzymology
  • Aspergillus / genetics
  • Aspergillus / growth & development*
  • Aspergillus / isolation & purification
  • Automation / methods
  • Biotechnology / methods*
  • Carboxylic Ester Hydrolases / biosynthesis
  • Genetic Engineering / methods*
  • Reproducibility of Results

Substances

  • Carboxylic Ester Hydrolases
  • feruloyl esterase