High-throughput screening of filamentous fungi using nanoliter-range droplet-based microfluidics.
Sci Rep
; 6: 27223, 2016 06 07.
Article
em En
| MEDLINE
| ID: mdl-27270141
ABSTRACT
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.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Aspergillus niger
/
Técnicas Analíticas Microfluídicas
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Alfa-Amilases
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Revista:
Sci Rep
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
França