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

TY - JOUR

T1 - Technical advance in fungal biotechnology

T2 - development of a miniaturized culture method and an automated high-throughput screening

AU - Alberto, F

AU - Navarro, D

AU - de Vries, R P

AU - Asther, M

AU - Record, E

PY - 2009/8

Y1 - 2009/8

N2 - 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.

AB - 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.

KW - Aspergillus

KW - Automation

KW - Biotechnology

KW - Carboxylic Ester Hydrolases

KW - Genetic Engineering

KW - Reproducibility of Results

U2 - 10.1111/j.1472-765X.2009.02655.x

DO - 10.1111/j.1472-765X.2009.02655.x

M3 - Article

C2 - 19486280

SN - 0266-8254

VL - 49

SP - 278

EP - 282

JO - Letters in Applied Microbiology

JF - Letters in Applied Microbiology

IS - 2

ER -