High-throughput screening of filamentous fungi using nanoliter-range droplet-based microfluidics

Filamentous fungi are an extremely important source of industrial enzymes because of their capacity to secrete large quantities of proteins. Currently, functional screening of fungi is associated with low throughput and high costs, which severely limits the discovery of novel enzymatic activities an...

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Published inScientific reports Vol. 6; no. 1; p. 27223
Main Authors Beneyton, Thomas, Wijaya, I. Putu Mahendra, Postros, Prexilia, Najah, Majdi, Leblond, Pascal, Couvent, Angélique, Mayot, Estelle, Griffiths, Andrew D., Drevelle, Antoine
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group UK 07.06.2016
Nature Publishing Group
Subjects
49/56
49/62
631/61
639/766/25
alpha-Amylases - genetics
alpha-Amylases - secretion
Aspergillus niger - enzymology
Aspergillus niger - genetics
Aspergillus niger - isolation & purification
Biotechnology
Enzymes
Fungi
High-Throughput Screening Assays - methods
Humanities and Social Sciences
Life Sciences
Microfluidic Analytical Techniques - methods
multidisciplinary
Mutation
Science
Yeast
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Summary:Filamentous fungi are an extremely important source of industrial enzymes because of their capacity to secrete large quantities of proteins. Currently, functional screening of fungi is associated with low throughput and high costs, which severely limits the discovery of novel enzymatic activities and better production strains. Here, we describe a nanoliter-range droplet-based microfluidic system specially adapted for the high-throughput sceening (HTS) of large filamentous fungi libraries for secreted enzyme activities. The platform allowed (i) compartmentalization of single spores in ~10 nl droplets, (ii) germination and mycelium growth and (iii) high-throughput sorting of fungi based on enzymatic activity. A 10 4 clone UV-mutated library of Aspergillus niger was screened based on α-amylase activity in just 90 minutes. Active clones were enriched 196-fold after a single round of microfluidic HTS. The platform is a powerful tool for the development of new production strains with low cost, space and time footprint and should bring enormous benefit for improving the viability of biotechnological processes.
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These authors contributed equally to this work.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep27223